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1、2015-12-30 1 I have just been made, where is my mother? Section I The birth of 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
2、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. I. Whats genetic engineering 1 . DNA is the key material for heredity 1)transfection of strepotococcus pneumoniae1)transfection of strepotococcus pneum
3、oniae 1928，F. Griffith，(18771941）, British, II. cornerstone of genetic engineering The experiment confirmed that the substance of heredity is DNA, NOT protein. The R type non-toxic living cells injected into mice, the mice without dying . The S toxic living cells injected into mice, the mice died of
4、 sepsis . The heating after killing S bacteria, injected into mice, the mice without dying . Mixed R non-toxic living cells with heat killed S, injected into mice, the mice died of sepsis （2）transfection of Phage（2）transfection of Phage In 1952, Alfred Hershy The secretion of A1 antitrypsin transgen
5、ic goat. 2015-12-30 6 The genes for the enzymes involved in the degradation of cellulose was cloned and expressed in Saccharomyces cerevisiae, it will be possible to produce glucose using cheap cellulose, fermented into wine. Exogenous gene transformation of Saccharomyces cerevisiae, produce quality
6、 beer. Or protein by Saccharomyces cerevisiae. 1. The development and utilization of cellulose 2. Wine industry II. Application of genetic engineering in industry With the recombinant plasmid of “super bacteria” decomposition of oil (Alkane), organic pesticide pollution. 1. Detection of water pollut
7、ion 2. Biodegradation Using the recombinant bacteria or transgenic fish to detect water pollution III. Application of genetic engineering in environment protection In 1976, 27 years old Robert Swanson of venture capitalist and University California professor Herb Boyer had a few beers, discussed the
8、 business prospects of genetic engineering. At the end of the discussion, they decided to build a company, and was named Genentech （Genetic Engineering Technology）. The first genetic engineering company was born in suspicious for the academic and business communities ! VI. Application of genetic eng
9、ineering in medicine. Manufacture of new vaccines (such as HIV, hepatitis B, hepatitis C, cholera, dysentery, SARS) 2. Produced by microorganism 3. Biological preparations designed high specificity 4. Development of a vaccine 1. The production of transgenic plants or animal for drug use Escherichia
10、coli or yeast to produce hormones (such as insulin), interferon Application of site directed mutagenesis technology design structure of proteins or enzymes, to produce biological agents with high specificity For example, the genetic engineering drugs America approved 2015-12-30 7 6. Medicolegal expe
11、rtise 7. Gene therapy The exogenous gene into the target cell to make up for the lack of target cells or mutation of the gene, or inhibit the abnormal expression of genes. For genetic diseases, cancer, cardiovascular disease, diabetes, etc. （ Is still in the stage of exploration ） Genetic engineerin
12、g of SCID treatment Patients of severe combined immunodeficiency (SCID) are immunocompetent, as long as bacterial or viral infection, will leads to the incidence of death. The mechanism of this disease is a mutations of chromosome encoding adenylate deaminase (ADA) gene. The treatment methods of gen
13、etic engineering could treat the disease. The gold rush on DNA track 2015-12-30 8 Because the purpose, species, biological materials, and experimental conditions of DNA extraction are different, there are many methods of DNA purification. One of the most commonly used is alkali extraction. plasmid D
14、NA Genomic DNA 2015-12-30 9 Plasmid DNA closed circular, not separated in the denaturation, renaturation quickly DNA double chain Denaturation DNA single strand Renaturation Alkali Neutral The linear DNA and chromosome or nicked plasmid DNA degeneration after double strand separation, to complex for
15、med by winding structure, combined with the protein SDS complex together, go on precipitation in the centrifugal time. Denaturation Lysozyme Hydrolysis the main chemical composition of cell wall peptidoglycan of 1, 4 glycosidic bond . In alkaline condition (pH8) is activated. Glucose Increase the vi
16、scosity of the solution, maintain the osmotic pressure, preventing DNA from mechanical force to degradation. EDTA The chelating agent of Mg2+ and Ca 2+ , can inhibit the enzyme activity, prevent DNA degradation by enzyme. NaOH-SDS NaOH： Alkali, alkaline conditions of the pH12, the DNA double strand degeneration SDS: The dissolution of cell membrane proteins and intracellular protein, synthesis “protein- SDS“ complex, so that the protein (including enzyme DNA) precipitat
