λ噬菌体和cosmid重组DNA克隆的快速限制性内切酶图谱分析方法  

Rapid Restriction Mapping of DNA Cloned in Cosmid or Lambda Phage Vectors

作　　者：柴建华[1] 

机构地区：[1]复旦大学遗传学研究所

出　　处：《Acta Genetica Sinica》1990年第2期136-142,共7页

摘　　要：cosmld克隆的线性化用λcos末端酶来完成,线性的cosmid或λDNA经部份限制性内切酶酶解后,分别与已标记的cos顺序探针杂交(探针为分别与λ的左端或右端的cos顺序互补的12核苷酸单链片段),杂交后的部份酶解片段经电泳分离和自显影后,酶切点位置可直接在X-底片上读出。在本实验室条件下,可一次完成二个克隆包括5—6种限制性内切酶的图谱分析,分析和作图可通过计算机或手工进行。A procedure for rapid restriction mapping of cosmid or lambda phage clones has been developed. The mapping of cosmid is based on linearization of circular cosmid DNA in vitro by the phage X terminase. Partial digestion products are selectively labelled at the right or left cos cohesive termini by hybridization with [32P] oligonucleotides complementary to the single-strand cos end. After gel electrophoresis and autoradiography, the restriction map can be directly determined from the 'ladder' of partial digestion products of cosmid or lambda clones with computer program or by hand.

关 键 词：Λ噬菌体 重组DNA 克隆 酶图谱 

分 类 号：Q785[生物学—分子生物学]