分子细胞学和基因工程上课课件pdf版

The S toxic living cells injected into mice, the mice died of sepsis . The heating after killing S bacteria, injected into mice, the mice without dying
1972, the first vector formed with the plasmid of small molecular weight and , amplicated in the bacterial cells.
4. competent cell
1970, M. Mandel & A. Higa found E. coli handled by CaCl2 could easily accepted phage DNA. 1972, S. Cohen found such cells also could accepted plasmid DNA.
In 1974, American National Institutes of Health (NIH) considering the potential danger of recombinant DNA, ask Paul Berg to form a recombinant DNA advisory committee. Committee consists of 11 members of the molecular biology and the recombinant DNA authority of the scholars. Commission published an open letter in July of the same year, requirements involved in the absence of clear recombinant DNA hazard degree and range, as well as before to take the necessary protective measures, suspended two test (with antibiotic resistance and tumor viruses and animal virus).
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Mixed R non-toxic living cells with heat killed S, injected into mice, the mice died of sepsis
The experiment confirmed that the substance of heredity is DNA, NOT protein.
1975, F. Sanger、A. Maxam & W. Gilbert invented the technique of DNA fast sequencing.
2. Boyer-Cohen experiment
1973, lab of S. Cohen of Stanford University finished recombination plasmid. R6-5 plasmid from E. Coli containing kanamycin resistance, plus another E. Coli plasmid pSC101，having double resistance. The first successful gene cloning experiment :gene fragment of Africa Xenopus laevis ribosomal , plus pSC101，transformation to E. Coli, corresponding mRNA successfully transcribed.
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I have just been made, where is my mother?
Section I The birth of genetic engineering
I. What’s genetic engineering
The formation of new combinations of heritable material by the insertion of nucleic acid molecules，produced by whatever means outside the cell， into any virus, bacterial plasmid or other vector system so as to allow their incorporation into a host organism in which they do not naturally occur but in which they are capable of continued propagation.
3. The central dogma and translation codon
1957, Crick, the central dogma
1964, Marshall Nirenberg & Gobind Khorana break 64 codons
III. techniques emerged for genetic engineering
（2）transfection of Phage
The R type non-toxic living cells injected into mice, the mice without dying .
In 1952, Alfred Hershy & Marsha Chase further confirmed that DNA is the substance for heredity.
3. The spread of cancer Occurrence area tumor viruses or other animal virus DNA into bacteria have the potential to expand cancer. 4. Artificial biological diffusion The newly formed DNA recombinant organisms accidentally diffusion may be potentially dangerous degree.
1. restriction enzymes
2. DNA ligase
1967, DNA ligase was discovered by 5 labs almost at theSmith, the first kind of RE HindⅡ isolated from haemophilus influenzae
1972, lab of Paul Berg in Stanford University accomplished the first recombination in vitro, putting DNA fragment of SV40 and phage together.
6. DNA sequencing
VI. The main content of gene engineering
1. Obtain of objective gene From complex genomes, by enzyme digestion and PCR amplification steps, isolated a gene fragment of DNA . 2. Recombinant preparation Insert the objective gene to the carrier molecules which could replicate themselves and with selective marker (such as antibiotic resistance) .
V. The character of genetic engineering
1. Cross species The exogenous gene to a different biological cells reproduce. 2. Clonal amplification Exogenous DNA can be amplified in host cells, and expressed with high level.
2. Experts attitude II. The safety standards of recombinant DNA research 1. Public concern In 1973, USA public first expressed concern about the application of recombinant DNA technology may cultivate new microorganisms having the potential danger, so as to bring consequences to human.
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In 1953, James D. Watson &Francis H. C. Crick showed us the DNA structure as the double helix at the first time, and the replication way of half-consreved.
II. cornerstone of genetic engineering
1 . DNA is the key material for heredity
1)transfection of strepotococcus pneumoniae