Chapter 5 Chapter 3 The Genesis Project - Interpretation of the Human Genome Project

Why do we want to spy on a small piece of text in our body, which Yan said can be surprisingly small?Do we have to be so arrogant?Why can't I let go of this text and not interpret it? "A good heart" is one reason, but "humility" is an even better reason, and "awe" wins.The evolutionary process took about four billion years to write down the DNA sequence possessed by synaptic organisms.And we're about to be able to read them, almost in the same way that we humans have invented books.What an astonishing prospect! The human genome is the genetic material of human beings (its chemical essence is deoxyribonucleic acid, referred to as DNA), including all gene sequences and non-gene sequences.Human genes are located on chromosomes, and there are three types, which are encoded as protein, transfer ribonucleic acid and ribosomal ribonucleic acid respectively. Among them, there are about 100,000 genes encoded as protein, which play a decisive role in various physiological functions and life phenomena. , is the most important type of gene.

It is now known that human life, old age, sickness and death, emotions, even ecological environment and biological evolution are closely related to genes.Therefore, the famous Nobel Prize winner in Physiology and Medicine, Du Bock once said: "The DNA sequence of human beings is the essence of human beings. Everything that happens in this world is closely related to this sequence, including human diseases including cancer. The occurrence is directly or indirectly related to genes..." Now the concept of gene is more specific, it is a sequence in DNA molecule that can express physiological functions.Because genes are most closely related to human life and survival, and out of human concern for themselves, the research and application of human genes has always been the center of gene research, especially in the medical field.Clinicians often encounter such a situation: the "ghost" of a certain disease lingers in some families, causing the same disease to occur from generation to generation within these families.

It is now clear that this "ghost" is the gene that causes the disease.Disease-causing genes are formed by mutations of normal genes.Because such diseases have obvious heredity, they are called genetic diseases.It has been found that there are more than 6,000 kinds of genetic diseases caused by pathogenic genes. They are also called single gene diseases, because the main pathogenic "ghost"-the pathogenic gene is only one.Clarifying the genetic principles of single-gene genetic diseases is a major breakthrough in genetic research.Later, people soon realized that virtually all human diseases are genetic diseases. In addition to the above-mentioned single-gene diseases, there are also polygenic diseases, such as malignant tumors, cardiovascular and cerebrovascular diseases, mental and neurological diseases, diabetes, rheumatism, Immune diseases, etc. In addition, there are acquired diseases, which are caused by Qingyuan microorganisms invading the human body, such as AIDS, hepatitis B, tuberculosis and so on.More importantly, scientists have discovered that no matter whether it is a single-gene disease or a polygenic disease, many genes are actually involved in the pathogenesis of the disease, but the role of the genes involved is primary and secondary, and there is a front and a back.This discovery made scientists fundamentally change the concept of gene research. They believed that in order to understand the mechanism of any disease or a certain health status, it is necessary to understand the genetic factors involved in the disease or health status from the genome level. The changing laws of all genes, not just the study of a certain Christian, gave birth to the great scientific project of the Human Genome Project.Scientists believe that the Human Genome Project is a major project in the history of human science along with the Manhattan Atomic Project and the Apollo Moon Landing Project.The program was first launched in the United States in 1990, and later scientists from Germany, Japan, Britain, France, and China formally joined in.

The emergence of the Human Genome Project is inseparable from the grounding of the "Tumor Project".The United States has launched the "Tumor Project" since the 1970s, but the investment at all costs has resulted in disappointing results.It is gradually recognized that various human diseases, including cancer, are directly or indirectly related to genes.Determining the base sequence of a gene is the basis of genetic research.At this time, scientists are faced with two choices: either to isolate and study several tumor genes from the human genome "piecemeal" or to fully sequence the human genome. In March 1986, Dubek published an article entitled "Turning Point in Cancer Research: Sequencing the Human Genome" in the American "Science" magazine. This short article was later called the "tender" of the Human Genome Project.Dubek said that the correct choice is to fully sequence the human genome, and such a large project should be completed by scientists from all over the world.

The "Human Genome Project" proposed by Du Burke with a far-sighted vision has generated huge repercussions around the world. Due to the large scale of the Human Genome Project, it has aroused widespread controversy in the United States. Some people say that the Human Genome Project is a game of taxpayers' money, and that the reduction of 3 billion bases is equivalent to 3 billion US dollars.At that time, even the sequencer used today had not yet been born.Therefore, some people say that we should start with small genomes, such as bacteria and fruit flies; or those with large economic value, such as pigs and sheep.

Furthermore, when the Human Genome Project appeared, other projects, such as the tumor project, cloning project, gene expression, neural activity research, etc., were all facing breakthroughs, so they were likely to become the "Oiran" of science and technology.However, in the heated debate, the Human Genome Project has been continuously improved, and the decision-making department has continuously considered the opinions of all parties, and finally believes that the Human Genome Project is the most important.Because people are the most important, and society cares most about people.In the process of evolution and fighting against diseases, human beings have accumulated a lot of their own genetic variation and diseases, which also provides materials for their own research; the problem of human genome has been solved, which can be directly used to solve other biological genome problems; In addition, the greatest economic value has been derived from the Human Genome Project.

In order to make the general public understand the Human Genome Project, the U.S. government has issued many pamphlets, such as "How Big is the Human Genome Project? Know Our Genes".Making the Human Genome Project the largest and most involved science popularization effort in U.S. history.The goals of the Human Genome Project have also been repeatedly discussed and drafted several times, and finally there are specific qualitative, quantitative, and timed goals for each part. In December 1984, Witt of the University of Utah was entrusted by the U.S. Department of Energy to host a discussion on recombinant DNA technology and the significance of determining the DNA sequence of the entire human genome. In June 1985, the U.S. Department of Energy proposed the "Human Genome Project"

preliminary draft. In June 1986, the feasibility of the Human Genome Project was discussed in New Mexico, followed by the US Department of Energy's announcement of the draft.At the Cold Spring Harbor Symposium in New York, Nobel laureates Gilbert and Berger hosted a meeting of experts on the Human Genome Project. In early 1987, the U.S. Department of Energy and the National Institutes of Medicine allocated a start-up grant of $5.5 million for the Human Genome Project. In 1987, the total amount was 166 million US dollars. In 1987, the United States began to prepare for the establishment of the "Human Genome Project" laboratory. In 1989, the United States established the "National Human Genome Research Center", with James Watson, Nobel laureate and proposer of the double helix structure model of DNA molecules, serving as the first director.

In 1990, the U.S. Congress approved the official launch of the U.S. Human Genome Project on October 1.Its master plan is to invest at least $3 billion in 15 years to analyze the human genome. In 1993, the United States revised this plan, and its main contents include: the construction and sequence analysis of the human genome genome map; the identification of human genes; the establishment of genome research technology; model organisms for human genome research; and the establishment of information systems.One of the most important tasks is the gene map construction and sequence analysis of the human genome.The most important are the following pictures: genetic map, physical map, and sequence map. The DNA sequence map is the highest priority and must be completed with quality and quantity.

In addition to the United States, other countries in the world have also started gene sequencing work.Notable is the United Kingdom. In February 1989, the UK began the Human Genome Project.It proposes a nationally coordinated, resource-focused mandate.Relevant laboratories across the country have uniformly obtained free experimental technology and experimental material services from the "British Human Genome Resource Center". Since 1993, the Sanger Center in London has become the largest sequencing center in the world, and it has independently completed more than 30% of the human genome sequencing task.

France's contribution to the Human Genome Project is around 3%.Its "National Human Genome Project," launched in 1990, was commissioned by the Ministry of Scientific Research to develop the National Academy of Sciences and Medicine. At the end of 1983, the Nobel laureate Dawsett established the Human Polymorphism Research Center with his own prize money.The French people donated at least $50 million.The Human Polymorphism Research Center and related institutions have made indelible contributions to genome research, especially the construction of the first generation of physical maps and genetic maps. Japan contributed 7% to the sequencing of the human genome.It started in 1990 under the impetus of the United States.In addition, Canada, Denmark, Israel, Sweden, Finland, Norway, Australia, Singapore, the former Soviet Union and East Germany have also started human genome research of different scales and with their own characteristics. China's Human Genome Project began in 1993 as a "major project" jointly funded by the National Natural Science Foundation of China, the National High-Tech Program, and the National Key Basic Research Program.The advisory committee of this project is composed of well-known geneticists, and the academic expert committee is composed of young and middle-aged scientists; there is also a "Chinese Genetic Diversity Committee" and a "Social, Legal, and Ethical Committee", and another secretariat is in charge International liaison, domestic coordination and daily affairs. my country is a populous country, accounting for 22% of the world's total population, and it is also a multi-ethnic group.my country's abundant population genetic resources are valuable materials for studying human genetic diversity, human evolution, and human-related disease genes.According to Academician Chen Zhu, Chief Scientist of the Southern Group of the Human Genome Project of China, "According to the actual situation in China, the initial goal of China's Human Genome Project is to make full use of China's rich genetic resources to conduct research on gene diversity and disease gene identification. In the past In the past few years, China has organized a group of high-level medical centers and national and departmental key laboratories in the field of genetics, established a nationwide network for the collection and preservation of genetic resources, introduced and established institutions including genetic and physical A relatively complete genome research system including maps, large-scale DNA sequencing, gene mapping, cloning, mutation detection, and bioinformatics. A number of important research results have also been obtained. In the field of genetic diversity, a multi-ethnic population The DNA sample bank has studied the genetic relationship of 30 ethnic groups and populations in the south and north of China, and compared them with 15 reference populations in the world. There is an obvious gene fusion between them; East Asian populations may have originated in Southeast Asia, while East Asian modern Homo sapiens and modern populations on other continents all originated "out of Africa" ​​100,000 to 200,000 years ago (by analyzing the mitochondrial In the study of DNA, the races of all countries have a relatively close relationship. Some foreign scientists believe that the ancestors of all modern humans came out of Africa 130,000 to 200,000 years ago and spread all over the world). On the other hand, disease genes Substantial progress has also been made in the research of hereditary high frequency deafness, and the chromosomal loci of several single-gene diseases have been located. In the study of the structure and function of genes related to leukemia and certain solid tumors.A number of achievements with international influence have been achieved.In recent years, preliminary success has also been achieved in the localization of polygenic diseases.In addition, some laboratories have also achieved breakthroughs in the study of human functional genes, and have obtained immunity from blood.There are more than 100,000 egys of neuroendocrine, cardiovascular system and liver, and more than 1,000 full-length tiDNAs of new genes have been cloned. " The "Human Genome Project" is to interpret all the genes on the human genome and analyze the four base pairs in the 24 chromosomal DNA molecules. 3 billion base pairs is a very long sequence. In order to better understand this long sequence, other auxiliary work is needed.In the "Human Genome Project", it is divided into two phases: the previous project of the DNA sequence map and the project of the DNA sequence map.Previous plans for sequence maps include physical maps, transcriptional maps, and genetic maps. The physical map of the human genome has two elements: sequence and location.In such a long sequence, the physical diagram acts like a map to signpost the various sequences.Through the method of molecular hybridization, using the complementary characteristics of DNA double strands, a DNA fragment hybridizes at this position, "it shows that the structure of this position is similar to it, which is the mark of this position, and this is the position marked by the sequence." The physical map also plays a more important role. With the sequence positions marked earlier, the cloned DNA fragments can be connected one by one.According to Professor Yang Huanming, the person in charge of the Chinese Human Genome Project, "If two cloned DNA fragments contain the sequence of a certain landmark, it means that part of the two fragments overlap. The DNA of our entire genome is composed of these overlapping The DNA fragments are all covered. In other words, these DNA fragments are representative of this region of our human genome, and the clones of these fragments are the experimental materials for our study of this region." The drawing of physical maps needs to be solved by means of genetic engineering. The technologies produced by genetic engineering over the years have been used in the production of physical maps.Chief among these are cloning techniques and molecular scissors. Simply put, cloning technology is the process of duplicating an individual using an individual's own self as a template without mating the parents. Molecular cloning of DNA uses the cells of organisms as carriers, and the cells themselves are cloned as well.The technology used now is DNA fragment cloning, that is to say, in plastic test tubes, fragments with a length of hundreds or even hundreds of thousands of base pairs are cloned, which is the basic technology of genetic engineering. Cloning of DNA fragments requires a "carrier". The earliest people used plasmids as the carrier of DNA fragments, which spliced ​​pieces of DNA together to achieve self-replication.Later, there were viral vectors, yeast vectors and bacterial vectors.Some have a large carrying capacity, some have stable functions, and some are easy to manufacture.Diversified means of DNA fragment replication. Gene splicing is the most basic method of genetic engineering. The long DNA molecules are the bonds, and the shortest chromosome 22 has 30 million fragments.How to cut them apart?This is where the molecular scissors come into play.The scissors are enzymes.It cuts DNA from the inside and recognizes specific sequences. There are two kinds of scissors, one is universal and can cut anywhere, and the other can only cut in special places. This kind of scissors is called restriction endonuclease.More than 300 endonucleases have been discovered so far.Interestingly, different incisions cut chains with different edges.In the DNA composed of two chains, some internal functional acids can only cut the fragments into flat ends, and some can cut and replace the ends, which is easy to reorder. In addition to enzymes that cut DNA molecules, there are enzymes that bind DNA molecules.With these tools in hand, gene splicing is possible. Another map to be completed by the Human Genome Project is the transcription map.We say that there are about 70,000 genes in humans.But among so many genes, only 1% to 5% of the genes are instructing protein coding.Because all kinds of life phenomena express and realize their functions through proteins.So grabbing these protein-coding DNAs roughly captures human genes, and that's what the transcriptome is going to do. All the DNA in each human cell determines nearly 100,000 genes, so only 10% of the DNA can be expressed in the cells of each tissue. Transcription is the first stage of expression. After DNA is transcribed, it becomes RNA with only one key. This RNA carries information, so it is called mRNA. RNA determines protein according to genetic code.Therefore, seizing these information-carrying mRNAs becomes an important task.It can be said that transcription map is the embryonic form of gene map. In the Human Genome Project, the partial sequences of DNA fragments are called expressible tag sequences. So far, in the Human Genome Project of international cooperation, these dJNA fragments have been found 1.6 million, and these 1.6 million have been analyzed and splicing, at least have represented the partial DNA sequences of tens of thousands of different genes. Because these genes in the transcription map are genes with expression function.Furthermore, the transcription itself is organized and time-specific, and it comes from a certain tissue at a known growth stage, so it can give information on the number, type, structure, and function of gene expression under normal conditions .In the future, we can also understand the expression of different tissues at different levels, different expressions, and different times, so that with normal and abnormal transcription maps, gene expression profiles can be constructed on this basis. Because this kind of transcribed DNA can provide reliable information for identifying which parts of the DNA sequence are coding DNA, and this is the most profitable and fastest-harvesting solution in sequence analysis. Diagnostic or gene cloning is used as a tool, so the construction of transcriptional maps and the scoring of fragments from the Sichuan team are highly competitive. A total of more than 400,000 dZNIA fragments have been patented in this area by private companies in the United States. Genetic map is an important tool to trace the essence of phenomena based on the principles of classical genetics combined with the progress of modern molecular biology. The essence of classical genetics is genetic analysis, which discovers genetic links between genes and manifestations.After genome analysis, it is found that a gene must have its locus in the genome, and this locus has at least two alleles, one is normal and the other is abnormal.If the abnormal gene is not expressed, the person is still normal, just a carrier.There is a problem of distance between this site and the genetic markers of the entire genome. If the site is close, exchange will occur, and if the site is far away, the frequency of exchange will be high.Scientists use a genetic marker to check whether the locus of this genetic marker has exchanged with the disease-causing locus in the family.The gene can be found at the corresponding distance of the genetic marker by means of the physical map.Although the cause of the disease is complex, it was possible to isolate the gene using a genetic map. Therefore, in the genetic map, the family is an important analysis object.Differences in sequence become the best "genetic markers." Physical maps, transcription maps, and genetic maps are all pre-sequence plans. The drawing of these maps is a preparation for the sequence map of the human genome. Only when the sequence map is completed can the sequence differences within the population be used as the genetic markers with the highest density To perfect the genetic map, so the sequence map is the most important part of the Human Genome Project. Chinese scientists participating in the Human Genome Project explained in detail how the drawing of the human genome sequence map was carried out in the book "Life Deciphered": The drawing of the DNA sequence map of the human genome can be compared as follows: Assuming that people only wear clothes of 4 colors, red, yellow, white, and black, the "Human Genome Project" is equivalent to the clothes worn by 3 billion people in the world. The clothes are all clear, and the order of location is indicated, such as the country, city, street, building, and room where they are located.The drawing of the human genome DNA sequence map is based on the above three maps, and adopts the "cloning to clone" strategy of "divide and win". Scientists use the markers that have been positioned to represent different regions in the human genome, that is, the "genetic markers" of the genetic map and the "physical markers" of the physical map, to find the corresponding "cloning of large DNA fragments" of the human genome.These clones are all known to overlap each other.Then use the machine to determine the DNA sequence of each clone, and then assemble them according to overlapping "adjacent fragment groups". In order to determine the sequence of these large DNA clones, these DNA clones should be determined in the genome according to the markers of the genetic map and the physical map, cut into small fragments of 1 to 2000 nucleotides long, and then "loaded" into a plasmid " Carriers, sent to bacteria for cloning, large-scale culture of bacteria, and the DNA of these "clones" were extracted from the bacteria. In my country's "Beijing Center", the staff have to prepare 5,000 to 10,000 clones of DNA every day as Sequencing "template".These DNAs must be pure in quality, accurate in quantity, and cannot be mixed with each other. Once the template is prepared, it is time to sequence.A first step is the "sequencing reaction".The method now used is the "enzyme termination method".To put it simply, a new strand is re-synthesized using the DNA to be tested as a template and marked with fluorescent substances of different colors.Like this, if a position of a section of sequence is A, just will represent the pyrochemical mark of A behind A, and so on.In this way, a new DNA chain with a length difference of one nucleotide is formed, and the last bit can be determined by the color of the fire: or ground or T, or motion or G. After the sequencing reaction is done, the second step is to analyze it on the 'Our Sequencing Protocol'.There are two main types of machines now, one is "gel electrophoresis" and the other is "capillary electrophoresis". They can separate DNA fragments with a length difference of only one base. The acid has been marked with fluorescent dyes of different colors, and the sequences of A, T, C, and G can be read out intuitively. These "sequences" are processed and checked by computer, and then some special computer programs are used to assemble the overlapping sequences.To determine the nucleic acid at each position, at least 5 to 10 measurements are required.If there are "holes" in the middle, these "holes" should be "filled" with various techniques, and finally a complete sequence of large fragment clones will be formed.These sequence fragments are then assembled according to the information of the "adjacent fragment group" to form a chromosome region and a complete chromosome sequence. Modern genome technology is a synthesis of molecular biology, genetics, genetic engineering technology, and bioinformatics.As the entire life science has entered the "sequence-based era", large-scale genome sequencing, assembly and analysis technology has become the most important "leading" and upstream technology of the bioindustry. The most concentrated expression of the management ability and human quality of scientific research enterprises. " It is not a fantasy that the cloning of a gene can form a gene industry.Most foreign pharmaceutical companies are investing heavily in genetics, and their scale is expanding very fast.For example, the gene transfer fee for an obesity disease is 140 million US dollars. As a result of the Human Genome Project, a human genome industry has emerged.By this stage, higher organisms are used as bioreactors.The genomes of these organisms are very complex. If foreign genetic engineering genes are put in, they will have an immune response and reject the foreign genes.However, there will be interactions between the foreign gene and the genes of its own genome, and the product protein of the foreign gene may also affect many reactions of the organism.Because some genes in the biogenerator need to be highly expressed, it is necessary to understand the entire genome of the bioreactor and understand the relationship between genes, thus generating the genome industry. Among the most established technologies in the genome industry is transgenic technology.For example, use cattle.Sheep and pigs are used as bioreactors to produce human erythropoietin.Production of human serum albumin by chickens.This is to connect the structural part of the human serum albumin gene to the chicken serum albumin, so that most of the original egg serum albumin becomes human serum albumin.Chickens that can produce human serum protein are naturally much higher than ordinary chickens. Transgenic technology can also turn cows into genetically modified cows that can be used to produce milk that is the same as human milk, and can also produce erythrotrophin.At present, only a few kilograms of erythropoietin are used in the world, but the annual output of a transgenic cow can reach one kilogram of willow fat.Currently transgenic cattle and sheep have extracted several human gene products from their milk. Transgenic technology can also produce human organs.In the history of medicine, the outstanding progress of the 20th century is the technology of organ transplantation.With the development of surgical technology and immune technology, humans can now perform various organ transplants.Such as cornea, kidney, heart, liver.Spare human organs have become a commodity in short supply, and tens of thousands of people around the world are waiting for organ transplants.In this case, scientists figured out how to transfer human organs to another animal.Let it grow human organs.Pigs were the first to be used for this kind of production, because pigs and humans are close relatives in evolution, and the structure of a pig's heart and kidney is similar to that of a human.About the same size, and only 10 months into the growing season.In the past, people were said to be stupid because of pig hearts, but now genetically modified pig hearts are going to be used for human organ transplants.Of course, to truly achieve this, we must also solve the problem of rejection of heterogeneous hearts. The pig’s heart must also be replaced with human ones, and the genes related to the development and function of the pig’s heart should not be replaced with human ones. In terms of plants, various types of genetically modified fruits and grains have appeared, and the United States has developed a "food genome". On the one hand, it is necessary to clarify the genomes of animals and plants; Said, to find new genetic resources.The green revolution started by human beings in the 20th century aims to find new species to produce enough food to feed all human beings. Now the food genome project will make people eat better.And solve population growth, relative lack of agricultural resources, ecological environment deterioration, and completely change the definition of farms. During the discussion on biological science at the Beijing High-tech Symposium in May this year, some scientists used vivid pictures to show the prospects of future fish production. The fish farms established on hundreds of hectares of land in the suburbs of Beijing, using genetic technology, can It satisfies the fish eating problem of tens of millions of people in Beijing. With the development of the Human Genome Project, the biochip technology, whose technical development direction was unknown before, has gained new impetus. In the process of serving the Human Genome Project, it has opened up broad prospects for its own development. Cloning technology is a biotechnology that has made breakthroughs in the past two years. British scientists cloned the first sheep, and other animals were cloned later.Yang Xiangzhong, a Chinese American scientist, said: "The impact of cloning technology on all aspects of life in the next century is in medicine and health. Therefore, some people in the United States compare biotechnology and atomic energy technology. But cloning technology brings benefits to human beings on the one hand, and on the other. Horror. What the Chinese care about is how the cloned people are counted in terms of seniority, and what the Westerners care about is that the cloned people are against God. Cloning technology is a technology that can reproduce a group of animals, plants and microorganisms. It turns out that we have inserted a root Branches can survive, which is a kind of cloning. In humans, identical twins are also a kind of cloning, because the genetic information is the same. Cloning has always existed in nature. It was not until 1997, when Dolly the sheep appeared in Britain, that people didn’t know about cloning. Know something and become a household name. It's a boon for animal cloning research." Yang Xiangzhong also said: A few years ago, it was proposed that in 2003, how many genes we humans have, the genetic map, and the location of the genes will be realized.A few months ago, scientists were making rapid progress on the human genome. Clinton said that by the end of this year, the mapping of the human genome would be completed, but within a few weeks, a company in the United States announced that the sequencing of all human genes would be completed within a few weeks. Positioning, we know that although the location of genes is very important, what is more important is what these genes are used for.Cloning technology will play an important role in studying the function of genes.Research genes that can cure diseases. Rare animal protection organization sent a letter to congratulate the science of cloning from animal skin cells, bringing good news to the protection of rare animals. Regardless of whether you are in favor of cloning or against cloning, cloning will definitely affect every aspect of human life in the future. In the future, the Human Genome Project will drive the development of other biological genome projects, such as the rice genome project that China has launched. Since humans and all animals, plants, and microorganisms are distantly related, the Human Genome Project is studying the highest and most advanced human. A whole set of strategies and techniques for the genomes of the largest and most complex organisms can be used to study the genomes of all other organisms. It is precisely because of this that the development of the biological revolution has changed the existing form of biological resources.Originally, to obtain a high-quality biological species, such as fine-bred cattle, or some kind of precious plant, it needed to be exported or smuggled.Now it is enough to use DNA technology, gene analysis and cloning technology to clone the DNA of this good cow or precious plant.As long as these cattle are sequenced, or only the relevant areas related to meat growth are measured, they can be sent back to China through the Internet.So far, some biological resources in our country have been lost silently. Genome sequencing technology upgrades individual biological resources to DNA resources and genome information resources.It is a new challenge for the protection and development of biological resources of every nation. Due to the huge financial interests of the gene, it immediately attracted many greedy eyes.The huge economic value of the Human Genome Project caused two countercurrents in the process of human genome research, which almost completely overturned the human genome research project.However, due to the righteous attitude of mainstream scientists who adhered to the spirit of the Human Genome Project, the Human Genome Project continued to accelerate in the struggle, and staged a fierce competition between mainstream scientists and private companies. The first battle of the Human Genome Project was the patent war.Patents have greatly promoted the development of modern science. They protect the intellectual property rights of scientists and ensure the public's understanding and application of inventions.However, the human genome is jointly owned by everyone, not the patent of any one person. Therefore, the famous human geneticist Fogel condemned "gene patents" as a nightmare for all mankind. Due to the huge investment in gene research, hundreds of millions of dollars will be invested in cloning a disease gene.Its potential commercial profits are also astonishing. According to the business logic of investment and return, some people apply for patents for genes with clear functions, but some people also apply for patents for genes with unclear functions.Now that 1,200 human genes have been patented, a fait accompli has been created.Issues can only be discussed on assignment fees and patent term. Because human genes are limited, if one is found, there will be one less. Therefore, the competition for genetic resources is a cruel competition. The loser can only use other people's patents for production.Without a patent, there is no right to produce. If gene sequences are also allowed to be patented, the consequences are serious.As a result of information monopoly, it can only be dominated by a few big companies, and anyone who wants to use this information must apply to him for permission. During the course of the Human Genome Project, there were two countercurrents.Both countercurrents were associated with the invention of new sequencers.We know that when the sequencing of the Human Genome Project began, there was not even a sequencing machine. In 1992, PE Company launched a new sequencer ABI373, which can read hundreds of nucleotide sequences at a time, so that EST can be analyzed on a large scale. So a scientist at the National Academy of Medicine applied for a patent, and it was supported by the former dean. In 1991, the US National Academy of Medicine submitted the first patent application for EST.This move was immediately opposed by mainstream scientists in the United States.Under intense pressure, the patent application was withdrawn.But a company in the United States, "Speed ​​Corporation" But with huge sums of money to support the scientist against the scientists of the World Genome Organization.Thus emerged two camps.At the same time as the moral debate, Spies confronted the scientists of the World Genome Organization by virtue of its economic strength.They are investing huge sums of money to speed up the production of patents, and have established their own EST databases to compete with public databases and lead several times.At the same time, sub-laboratories for EST production are established in some third world countries. Scientists from the International Genome Organization also isolated EST at the fastest speed, increasing the number of EST sequences in the public gene bank, and put the measured sequences online as soon as possible, resulting in a public fait accompli.At the same time rely on legal means, tools of public opinion, and moral force to fight for reason. In the patent application, there are also twists and turns.In 1997, the US Patent and Trademark Office considered granting a patent to IISI'. Public opinion was uproarious.The National Institutes of Medicine strongly disagreed. In 1994, the US National Academy of Medicine withdrew nearly 7,000 EST patent application pairs, clearly stating that incomplete or complete gene sequence applications with unclear functions and practical significance are not in the best interests of public health and science.There is a general resistance to the PTO's definition of the utility value of ESTs, arguing that this will cause problems for scientists to enjoy early access to sequence information. The World Genome Organization strongly urges the Patent and Trademark Office to withdraw the Egr patent decision.Some scientists in the United States broke with Spies.All sequences will be made public as soon as they are determined, and the amplification speed of the public database remains normal. 1998年，随着新的测序工具的出现，出现了第二次逆流。5月三叉口，PE公司推出了新一代的“毛细管测序义'，使测序的工作提高了许多倍，自动化程序也高级得多。 但是这家公司首先将300台机器自己使用，并且投资3亿美元，让原来那位美国国家医学研究院的科学家专门成立了赛来拉公司，号称要在3年内用新的方法完成人类基因组计划的全部序列。他们说，他们只要几百个基因的专利，但是在发布的时间上和对序列的垄断上他们采取拖延的态度。他们拒绝在24小时内公布所有数据，表示要把数据分析完了，选择出最重要的基因为自己所用，然后对这些选择出的基因索价年使用费至少为500万美元。 国际基因组组织的科学家奋起抵制这一做法。他们认为，赛来拉公司的新策略所用的基因装配，用的是全球科学家几十年心血的遗传图、物理图，他们免费从国际公开数据库中得到这些信息，本身就不公平。再者，如果国际基因组计划就此流产。10年的心血将付之东流，许多人才会流向赛来拉公司。他们将在这个领域内肆意妄为。因此，他们一方面是计划得以继续，向政府申请经费，加速研究，与赛来拉公司竞争。 据美国《时代周刊》的一次民意调查，72％的民众不赞成几个公司“自己出钱”，完成人类基因组测序并专利重要基因，美国国会经过多次激烈辩论，听取了主流科学家的意见，保证了对HGP的继续支持。 值得注意的是，曾蓄意破坏HGP，孤立美国主持正义科学家的PE公司，专门派出各种身份的人，到有意参与“人类基因组计划”的发展中国家游说，也同样到过中国。说什么“即使全球合作，也都斗不过我们公司”，“参加测序，等于白花钱”，“数据反正是白用的，干嘛还花钱参加”……居然真影响了一些国家的决策者。 争论还在继续，1999年9月1日，在“人类基因组计划”有关“工作框架图”的最后一次策略会议的前两天，“赛里拉”居然宣布自己已完成人类基因组测序的印％，它的股票一日暴涨切美元，上升幅达四．5％。实际上他们的数据，是目前难以装塔的原始数据，离“工作框架图”相差甚远。但从另一方面，却反映了民众对人类基因组数据潜在价值的认同。 尽管“人类基因组计划”的所有资助者、所有参与的实验室，都一致同意并许诺： “人类基因组计划”的数据，应该“平等、免费”分享，并签订了“百慕大原则”：所有数据都应在24小时内公布，但“赛里拉”等公司肯定不会就此罢休，因为，“人类基因组计划”的科学意义与经济意义实在太重大了，争持不会停止。 人类基因分离与研究的最终价值是人类疾病的预测、诊断与治疗；基因功能的鉴定，关键是与疾病等表型的联系，这就决定了基因分离一应用这两头都需要与疾病挂钩。疾病家系、人群、患者的遗传材料，成了具有科学、经济意义的基因资源。而拥有这一资源的发展中国家，成了争夺基因的“狩猎场”。 印度科学家首先挺身而出，给印度政府施加压力，要求以行政措施保护印度的基因资源。我国基因资源外流情况，比起印度，有过之而无不及。 据《今日生物世界》报道，美国西夸纳公司已取得中国一个很大的哮喘家系，随后多次宣扬这一家系的价值，以配合宣传他们的工作。中国这一家系的外流，国内学术界至今仍蒙在鼓里。 要说基因资源，中国是首富。一是中国的人多，病也最多；二是中国人几代同堂，没有天灾人祸不动窝，少数族群多生活在偏远的大山里，形成的家系最多最纯。一些基因资源掠夺者把目光投向了中国。 据美国权威的《科学》杂志1996年报道：哈佛大学“群体遗传学计划”，要在中国研究包括糖尿病、高血压、肥胖症、早发心脏病、关节炎、精神分裂症与传染病在内的几乎所有“文明病”。这一计划要用2000万中国人的血样及DNA样本，因为中国可提供廉价研究材料；巨大的人口可以使科学有鉴定功效细微的基因。这一计划要通过6个中国医学中心，而这些中心（不管刚挂牌，或还没有挂牌）的正主任则是美国这一项目的负责人，他们声称将扩大经费，而多个药物公司赞助的筛选600万中国人以研究哮喘基因的项目已经上马。 中国预防医学科学院与美国BMI等公司合作，以研究“膳食、生活方式和慢性消耗性疾病的关系”为题，在我国收集血样与有关流行病学方面的资料，计划采集50万人的血样与所有个体的体检、临床数据。此协议写道：“本项工作产生的全部知识产权全部归BMI所有，包括版权、专利。商标注册。”中方明确声明：“本项目所涉及的知识产权将为BMI所有。”为了吸引投资者和遵循国际商业惯例，此点必须在协议中清楚地表明。由于在这一商业活动中，中方并未投资，因此在知识产权方面不可能要求平等。 印度政府正计划通过立法，来限制外国研究者与药物公司获取该国的生物资源。如果没有“国家生物多样性管理委员会”批准，将具商业价值与生物资源有关的标本、数据输入国外，将受严厉惩罚，处以5年的监禁或3万美金罚金。 冰岛国会为制止掠夺遗传资源的“生物海盗”，反对“直升飞机式研究”或“取了血样便跑”的研究，立法制止人类组织样品出口。外国公司要想以冰岛人群进行研究，只能在该国国内进行，或保证让冰岛“免费”享受所有研究成果。 美国政府通过外交途径，转告外国政府：美国人在美国以外采集人类基因资源，并没有得到美国政府的批准与资助，只是个人行为。 联合国教科文组织于1995年成立了“国际生物伦理委员会”。经3年讨论，反复修改，起草了《关于人类基因组与人类权利的国际宣言》，简称《人类基因组宣言人1997年三三月五三日，经“联合国教科文组织”第二十九届会议通过。1998年11月27日又经“联合国大会”批准，成为名副其实的国际文件。 《人类基因组宣言》是历史上第一个有关科学研究的宣言。它的发表，充分反应了“人类基因组计划”可能对科学、经济、伦理、法律及社会方方面面的影响，以及就这些问题讨论的迫切性与严肃性。《植言》被比拟为1948年《人权国际宣言》与《纽伦堡法典》，其宗旨是保护人类的基因组。 《人类基因组宣言》有4条基本原则：人类的尊严与平等，科学家的研究自由，人类和谐，国际合作。 为保护发展中国家的权益，《人类基因组宣言》在历史上第一次提出与发展中国家进行国际合作、“南北合作四大原则”。由于本书（指《生命大解密》）作者授权解释与此有关的条文，因此特别强调这一部分。 1.全面解释人类基因组研究的风险与利益，防止滥用。人类基因组的研究是把“双刃剑”，由于科学认识暂时的局限性以及对技术的渴望，发展中国家的决策者与民众有可能忽视风险，在立法方面滞后。因此，决不允许以“帮助发展”、“技术领先” 的名义，在发展中国家进行在发达国家不允许的实验，转嫁风险，把发展中国家作为“侏罗纪公园”的实验场。 2.以提高发展中国家进行人类生物学与遗传学研究能力为宗旨。考虑到发展中国家的特殊问题与需要。特别是解决他们的研究能力，不能搞假合作、真掠夺特殊人群与患者的遗传材料。 对于发展中国家，提供特殊材料，是一个民族对科学发展的贡献，不能“将物自居” 拒绝国际合作，而耽误这一疾病的研究。“资源换技术”，在迫切需要技术的时候是可以考虑的。但国际间的合作，要考虑是否有助于提高本国的研究能力。本国的科研人员，有对自己的资源进行开发、利用的优先权。 在发展中国家取得资源，一定要服从该国该地的有关法律与管理法规。特别要尊重“知情同意”的权利，不能利用当地人民对科学暂时的不了解，以任何名义、手段，甚至与地方当局合作，从当事人那里骗取遗传材料。 3.国际合作应有利于发展中国家对科学技术成果的分享。由于历史原因、经济能力的限制，发展中国家对人类基因组研究没有投入，在研究规模与速度上比不上发达国家，甚至至今还没有参与，但人类基因组是全人类基因遗产与财产，因此，这绝不能影响发展中国家分享“人类基因组计划”成果的权利。只有这样，才能促进全人类的和睦和整体进步。 4.促进发展中国家与发达国家自由交换科学知识与信息。科学家相互自由交换科学知识与数据，是科学发展的根本保证，也是科研与应用的基本保证，由于人类基因组信息对全人类的重要性，更突出了自由交流的意义，保证不扩大发达国家与发展中国家在科学上的差距。 《人类基因组宣言》事关全球各国，影响千秋万代。因此，“联合国”要求所有成员国采取措施，通过各种手段，向民众、特别是科学决策者传播《宣言》的原则条文，提高整个社会对基因研究中可能涉及人类尊严的关切程度，促进这些原则的实施。 中国在国际人类基因组计划中承担了1％，这是人类3号染色体短臂上的一个约30MB区域的测序任务，该区域占整个人类基因组的1％。说起这个计划的实施，没有人能比参与这项工作的中国科学家更有发言权，现在就看看他们对这件事情的全面描述： 中国参不参与序列图绘制的国际合作，已讨论了10年。如果认同人类DNA序列图是“重中之重”，关系到21世纪我国生命科学与生物产业的基础建设，不参与序列图绘制，这一步拉开了，将眼巴巴地使我国永远失去参与的机会。 苦头我们已开始吃了，如对虾病毒基因组测定忍痛让人。一步被动，势必长期被动，全局被动，耽误国是。历史将证明，中国建立大规模的基因组序列图构建系统，只是时间的问题。越晚，我们民族付出的代价就越大。不做，就是我们的失职。历史将要追究所有人的责任，包括讨论中持不同意见的双方。因此，我国的决策部门，所有相关的研究人员，一直在沉重地、痛苦地思考这个问题。 最终中科院遗传所的“人类基因组中心”（简称北京中心）于1998年8月4日开张。 1999年2月决定搞大规模基因组测序，4月预运行，以创造加入“国际测序俱乐部”的条件。7月7H在国际人类基因组测序协作组登记，申请加人“国际测序俱乐部”。 1999年9月三日。在伦敦举行的第五次人类基因组测序战略会议上，作为新的成员，北京中心与已为人类基因组做出的卓越贡献的万个中心一起讨论战略，商议标准，界定区域，分析面临的问题，一起分享喜忧。占世界人口20％的中国，负责测定人类因基组序列的1％。 “国际测序俱乐部”听取了北京中心关于实验室面积。设计规划、设备类型及实际运行情况的数据统计；人员组成及素质、技术培训与实际运作等方面的情况介绍，以及依据设备、试剂、人员的实际投入与产出等所有数据做出的详尽预算。北京中心自信地宣布：保证中国科学院及其遗传所。中国中央政府及其他有关部门、地方政府和其他各种来源及中国民众对这一项目的财政支持，全额经费绝对能及时到位。 滴水穿崖，非一日之功。北京中心的关键设备运行情况与国际同行并驾齐驱，令人信服地说明中心人员已掌握全部的技术关键与细节，以及世界级中心的管理与动作。北京中心自豪地展示了自己测定的难度最大，投入最大，意外最多，准确率最高的区段，以及已递交的4个片段628Kb数据。这些数据，已使中国成为递交人类DNA序列数据最多的6个国家之一。北京中心对与国际同行同步，即在2000年春求完成“包干”区域的测序充满信心，并保证—半以上的序列达到“终围”的质量标准。 北京中心动最后表示：在研究过前四次“战略”会议文件的基础上，保证俗守HGP精神，特别是有关数据的即时公布与免费分享的原则。北京中心还重申反对人类基因组基本信息专利的立场，保证不保留任何数据，不申请类似的专利。 由于中国注册较晚，原定会议程序并未列入。为中国代表能及时与会，北京中心几位国际顾问四处联系，出谋划策。主要负责人最后通过越洋电话，当即决定邀请中国代表与会，各国代表纷纷祝贺。会场上数位代表或以旧交之了解，或以目睹之事实称赞中国的进步。HGP精神感人至深。在国内，基因组学界的前辈们谆谆教导，有关领导语重心长：“志在必得”兄弟院所真诚合作，地方政府鼎立支持，同仁同道倾囊相助。 截至1999年2月13日，北京中心已投入了28万余个测序反应，已完成申报投入50万个反应的一半以上，累计测定了110Mb的序列，相当于把这一区域测了3次。按国际标准，已递交国际数据库16．SMb的一致性序列，已完成了“工作框架图”的55％。2000年春求完成“工作框架图”完全有把握。国家人类基因组北方与南方中心也参与了这一项目。 除了完成“工作框架图”的任务外，北京中心已建立了完成单个BAC的序列组装的能力，已完成组装并递交了6个完整的BAC的DNA序列，合计长度的0．gMb。并建立了将几个BAC的序列组装成一个连续片段的能力，能将10个BAC克隆序列组装成三．IMb的大片段。另外，建立了数据分析与基因鉴定的能力。24小时内，在内部网络中完成全部数据的初步分析。 “1％项目”尽管还会有争议，但它的意义，已逐步显示： 1、显示了中国领导人与决策者的高瞻远瞩与英明果断。 中国参与国际“人类基因组计划”，正如国际同行与海外留学生所说的，充分显示了我国新一代领导人与决策者，对全球科技格局的了解和参与国际合作重大课题的新思维与新策略。中科院以1000万人民币的投入，创造了又一个“中国第一”，成为中国的HGP中最具影响与实际产出最明确的主要部分。而我国以500万美元的投入，进入五强国历时10年、总投资达又亿美元的HGP行列。这种在关键时刻所表现的远见卓识、决策的果断与经费到位的快速，都是前所未有的。 2.改变了国际人类基因组研究的格局，提高了人类基因国际合作的形象，受到了国际同行，特别是参与“人类基因组计划”的各个中心以及发展中国家的欢迎和称颂。 国际合作、公众支持的“人类基因组计划”已历时10年，一直受到美国私人公司的挑战。国际“人类基因组计划”负责人一直希望提高国际合作的形象，而由于'人类基因组计划”投入巨大、技术复杂，使英、美、日、德、法之外的其他国家，望而却步，采取了“你成我拿，你干我看”的观望态度。多数国家，特别是发展中国家，一方面基于HGP对人类本身的认识与生物产业发展之攸关，在道义上力争共享人类基因组序列信息的权利；一方面由于不能直接参与而没有实际的发言权。 正在这个时候，中国的参与，无疑受到国际“人类基因组计划”团体与发展中国家的一致欢迎。真正参与并分担实际任务的其他15个中心的负责人，无不致电致信表示欢迎，至诚之心溢于言表。 在联合国教科文组织“国际生物伦理委员会'第六次会议上，我国委员，本书（指《生命大解密》）作者之一的杨焕明先生，作为发展中国家惟一真正参与“人类基因组计划”的代表，深感地位的改变：国际“人类基因组计划”负责人，希望我国代表多为HGP精神呼吁；发展中国家又希望我国能在“人类基因组计划”中多为他们争取权益。 “人类基因组计划”接近完成，各个国家讨论“人类基因组计划”的层次已达到国家最高领导人，我们这一地位将更加令人注目。 3. “1％项目”，使我国理所当然地分享“人类基因组计划”的全部成果与数据、资源与技术，拥有有关事务的发言权。 国际“人类基因组计划”的宗旨是全球合作的跨国项目。不参与，就不能直接获得资源与技术；不参与，就不可能有发言权；不参与，就随时有失去分享数据的危险。 北京中心数次派人员直接到美国最大、自动化程度最高、生物信息学最为先进的中心参观学习。所到之处，无不受到主要负责人亲自接待，具体介绍5一6个小时，并帮助培训掌握最核心的技术。我们测序的所有BAC克隆，都来自国际标准化文库，所有的BAC信息都是由他们免费提供的。 国际“人类基因组计划”所受到的挑战，带来了我国在“人类基因组计划”有无发言权的问题。如这次与美国私人公司“赛里拉”的谈判，事关世界各国能否继续分享人类基因组序列信息的大局。国际人类基因组中心的负责人向我们通报了情况并诚言：我们向您许诺，没有经过您参加的会议的七寸论，我们不会做出任何决定。如果没有“丑项卧'，这种情况是不可想象的。 4.建立了我国自己的、接近世界水平的基因组研究实力。 通过参与而分享了国际人类基因组的资源与技术，我国在短短的6个月内，走过了别人积累10年的历程，缩短了可直接比较的差距。中国科学院遗传研究所人类基因组中心日产数据ZMb，相当于世界上最强的两个中心在1993年的年产量。我们已建立了一支训练有素的能打硬仗的队伍，建立了有自己特点的全套系列与技术，积累了世界级大规模中心的运作管理经验，奠定了进一步发展的坚实基础。 我国信息产业的上游——软件与硬件，已受制于人，我们民族已在为此付出代价。 资源基因已成为一个国家发展的战略资源。争夺这一资源的“世界大战”已经打响。 2000年1月13日，企图垄断基因组信息的赛里拉公司宣布：在中国的台湾与上海同时登陆。在台湾，他们得到了政界首要的支援，计划投资一亿美元。在上海，他们收购了原先以。'“测序服务”注册的外资公司Gewt的95％的股份。赛里拉公司的头头公开声称：得到中国富甲天下的动物、植物与人类的遗传的多样性资源，是赛里拉公司扩大国际商务与基因组信息的基础。无疑，赛里拉公司此举是有“远见”的：一方面，以掌握了中国丰富的生物资源为宣传，直逼坚持“平等分享”原则的国际“人类基因组计划”；另一方面，以雄厚的资本：一期投资3亿美元，现股票市场估价为80亿美元，与强大的技术实力：拥有300台毛细管测序仪，号称“天下第三”的超大型计算机，妄图实际控制中国的生物资源的梦想。 中国的信息产业已失“源头”又缺实力，正在走以市场换技术求发展的路子，我国的生物产业，再也不能重蹈“以资源换技术”的覆辙。 通过“1％项目”，我们已完全具备与赛里拉等公司，在速度与成本上的竞争能力。 “1％项目”已成为对付“赛里拉”掠取我国生物资源的桥头堡。 建立我国的基因组测序的强大实力，与“赛里拉”等公司抗争，是保护、发展、利用我国丰富的生物资源的重要前提。由于参与了“人类基因组计划”，随着“1％项目” 的完成，我们与“赛里拉”的抗争，不但不会影响我国的开放形象，相反，会进一步增进国际同行的理解与信任。 “1％”为21世纪的中国生物产业带来了光明和希望；历史将证明“1％”在中国科技史上的意义。（见杨焕明等着《生命大解密》） 一功能基因组学 当前，随着结构基因组学向染色体完成序列图的目标顺利进展，以揭示基因组的功能及调控机制为目标的功能基因组学已提上议事日程。美国HGP（人类基因组计划）1998一2003年的新目标，除了完成基因组全序列测定之外，还有人类DNA序列变异，全长d3NA克隆、发展全基因组水平功能分析的技术、模式生物体等属于功能基因组学范畴的内容。一般认为功能基因组研究包括的核心科学问题有：基因组的多样性；基因组的表达及其时、空调节；模式生物基因组研究等。还应指出，生物信息学也是对功能基因组学数据进行储存、分析和发掘的基本手段。 1.基因组多样性的研究 人类是一个具有多态性的群体。不同群体和个体在生物学性状以及在对疾病的易感性航性上的差别，反映了进化过程中基因组与内、外环境相互作用的结果。开展人类基因组多样性的系统研究、无论对于了解人类的起源、进化和迁徙，还是对于生物医学均会产生重大的影响。已知人类基因组DNA序列中最常见的变异形式是SNP，在全基因组中估计有3—10X10个。与罕见的单碱基变异所不同的是，SNP等位位点的频率应等于或高于1％。当SNP位于基因的编码序列中即称为cSNP。若CSNP引起蛋白质重要部位氨基酸的变异，可导致其功能改变；位于基因调控序列中的SNP则可能影响基因表达的剂量。故这两种SNP的生物学意义更为显着，是基因组中决定人类表型多样性的核心信息。另一方面，SNP因连锁不平衡（LD）所形成的单倍型，也可用于关联研究来确定与之连锁的生物学性状相关序列。目前，已发展了多种自动化和批量化检测SNP的技术，其应用范围十分广泛，包括连锁分析与基因定位；疾病的关联研究；多基因疾病的基因定位；个体识别和亲子鉴定，发病机理的研究；以及研究生物进化，生物间相互关系等。前曾述及，edera的人类基因组测序计划包括了SNP。针对此种情况，1999年国际上10家大药厂与三家研究机构建立了SNP协作组，以后又吸引信息技术产业的数家大公司参与，对24个来自世界不同群体的个体，进行SN'P的随机筛选，计划到2001年获得30万