第十三讲-2006基因工程.pptVIP

Genetic engineering and biotechnology 用rBST处理后，牛的奶产量通常增加15%，这对牧场经营者来说是有利可图的。 但注射rBST可以引起局部肿胀，加重脚部感染、乳腺炎，并给牛的繁殖能力带来影响。 Administering rBST can produce localized swelling at the site of injection, and can exacerbate problems with foot infections, mastitis and reproduction. Counter-argument---赞成rBST的观点认为上述问题在没有注射rBST的牛群中也存在 天然的或重组的生长激素可以增加胰岛素样生长因子（insulin-like growth factor, IGF-1）的水平，进而增加牛奶产量。 有证据表明IGF-1可以促进癌细胞的生长。所以使用rBST可能对健康带来危害。 但赞成rBST的人认为哺乳期早期阶段的IGF-1水平比哺乳期开始100天后注射rBST后牛的IGF-1水平要高。 There is evidence that IGF-1 can stimulate the growth of cancer cells. 反对者认为： rBST与治疗性蛋白不同，后者只给少数病人使用。 而牛奶是大部分人的消费品，不管rBST引起的危险多么微不足道，都是不能接受的。 Those who oppose the use of rBST point out that, unlike a therapeutic protein that would be used for a limited number of patients, milk is consumed by most people, and any inherent risk, no matter how small, is therefore unacceptable. 第二节 蛋白质工程 protein engineering ? 蛋白质工程：通过改变基因序列来改变蛋白质结构。 主要使用离体突变技术。 离体突变技术可以在基因序列中引入特定的突变。 Altering the structure of proteins via alteration to the gene sequence, and has become possible due to the technique of mutagenesis in vitro. Mutagenesis in vitro enables specific mutations to be introduced into a gene sequence. 定点突变，也称寡核苷酸介导的突变： 1，用M13噬菌体制备单链模板，含需要改变的基因。 2，合成15-30个核苷酸长度的寡核苷酸片段，该片段与待突变的模板DNA区域互补，中间含突变碱基。 Oligonucleotide-directed or site-directed mutagenesis: Single-stranded template (ss DNA)-----containing the gene to be altered.-----produced via M13 cloning system. An oligonucleotide (usually 15-30 nucleotides in length)，complementary to the region on interest, and has desired mutation in the middle. 3，模板与寡核苷酸退火。 4，突变位点错配，但两侧的互补序列可以起到稳定作用。 The template and oligonucleotide are annealed. the mutation site will mismatch, but the flanking sequences will confer stability. 5，用DNA聚合酶和DNA连接酶合成和连接产生双链DNA分子。 6，导入大肠杆菌复制，产生两个子代分子，其中一个的靶位点已经发生改变。 the template is then copyed using DNA polymerase. This gives rise to a ds DNA which, on replication (when introduced into E. coli), will yield two da