生物技术专业英语第4课.doc

Lesson 4 LIGATION, TRANSFORMATION AND ANALYSIS OF RECOMBINANTS DNA ligation: T4 DNA ligase repairs breaks in a dsDNA DNA backbone and can covalently rejoin annealed cohesive ends in the reverse of a restriction enzyme酶 reaction, to create new DNA molecules. DNA 连接：T4 DNA连接酶在双链DNA的分水岭处修复破损的地方，然后在反向限制酶反应中，能共价地再加入退火的粘性末端，从而形成新的DNA分子。 Recombinant DNA molecules: The use of a restriction enzyme, followed by DNA ligase, can create recombinant plasmids, with a target DNA fragment inserted into a vector plasmid. 重组DNA分子：限制性内切酶的应用，紧随DNA连接酶，能通过将目标片段插入一个人载体质粒形成重组质粒。 Alkaline phosphatase: Treatment of the linear vector molecule with alkaline phosphatase will remove the 5-phosphates and render the vector unable to ligate into a circle without an inserted target, so reducing the proportion of recreated vector in the mixture. 碱性磷酸酶：碱性磷酸酶对线状载体分子的处理是移除5’的磷酸键和会使没带出入目标基因的载体连接成一个环，从而混合物中减少重新形成的载体的比率。 ransformation: Transformation is the process of take-up of foreign DNA, normally plasmids, by bacteria. Plasmids are cloned by transfer into strains of E. coli with defined genetic properties. The E. coli cells can be made competent to take up plasmid DNA by treatment with Ca2+. The cells are plated out on agar and grown to yield single colonies, or clones. 转换：转换是提取外来DNA的过程，一般是在细菌里的质粒。带有明确遗传特性的质粒是通过转移到大肠杆菌菌株才被克隆的。大肠杆菌细胞能通过Ca2+ 的处理提取质粒DNA。细胞被平铺在琼脂上，然后逐渐产出单一的菌落或是克隆体。 Selection: Bacteria which have taken up a plasmid are selected by growth on a plate containing an antibiotic to which the plasmid vector encodes resistance. 筛选：已经提取了质粒的细菌是通过含有看抗生素的生长培养基选出来的，而这种抗生素是阻碍质粒编码的。 Transformation efficiency: The efficiency of the transformation step is given by the number of anti-biotic-resistant colonies per microgram of input plasmid DNA. 转换效率：转换步骤的效率是通过输出质粒DNA每微克抗生素抗性菌落的数量所给定的。 Screening transformants: In many cases, such as when using DNA libraries, plasmid and other vectors have been designed to facilitate the screening of transformants for recombinant plasmids. In the case of a simple subcloning