分子细胞学和基因工程上课课件版

2015-12-301I have just been made, where is my mother?Section I The birth of genetic engineeringThe 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.I. Whats genetic engineering1 . DNA is the key material for heredity1)transfection of strepotococcus pneumoniae1)transfection of strepotococcus pneumoniae1928，F. Griffith，(18771941）, British, II. cornerstone of genetic engineeringThe 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 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 PhageIn 1952, Alfred Hershy The secretion of A1 antitrypsin transgenic goat.2015-12-306The 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 beer. Or protein by Saccharomyces cerevisiae.1. The development and utilization of cellulose2. Wine industryII. Application of genetic engineering in industryWith the recombinant plasmid of “super bacteria” decomposition of oil (Alkane), organic pesticide pollution.1. Detection of water pollution2. BiodegradationUsing the recombinant bacteria or transgenic fish to detect water pollutionIII. Application of genetic engineering in environment protectionIn 1976, 27 years old Robert Swanson of venture capitalist and University California professor Herb Boyer had a few beers, discussed the 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 engineering in medicine.Manufacture of new vaccines (such as HIV, hepatitis B, hepatitis C, cholera, dysentery, SARS)2. Produced by microorganism3. Biological preparations designed high specificity4. Development of a vaccine1. The production of transgenic plants or animal for drug useEscherichia coli or yeast to produce hormones (such as insulin), interferonApplication of site directed mutagenesis technology design structure of proteins or enzymes, to produce biological agents with high specificityFor example, the genetic engineering drugs America approved2015-12-3076. Medicolegal expertise7. Gene therapyThe 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 engineering of SCID treatmentPatients 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 genetic engineering could treat the disease.The gold rush on DNA track2015-12-308Because 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 DNAGenomic DNA2015-12-309Plasmid DNA closed circular, not separated in the denaturation, renaturation quicklyDNA double chainDenaturationDNA single strandRenaturationAlkaliNeutralThe linear DNA and chromosome or nicked plasmid DNA degeneration after double strand separation, to complex formed by winding structure, combined with the protein SDS complex together, go on precipitation in the centrifugal time.Denaturation LysozymeHydrolysis the main chemical composition of cell wall peptidoglycan of 1, 4 glycosidic bond . In alkaline condition (pH>8) is activated. GlucoseIncrease the viscosity of the solution, maintain the osmotic pressure, preventing DNA from mechanical force to degradation.EDTAThe chelating agent of Mg2+ and Ca 2+ , can inhibit the enzyme activity, prevent DNA degradation by enzyme.NaOH-SDSNaOH： Alkali, alkaline conditions of the pH>12, 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) precipitation.Acetic ac