绛红色小单孢噬菌体MPh1在大肠杆菌中有启动子功能的DNA片段的克隆及顺序测定  

Cloning,expression and sequencing of a DNA fragment having E,coli promoter funcation from phage Mph1 attacking Micromonospora purpurea

作　　者：周健明[1] 陈孝康[1] 杜艳[1] 王建红[1] 汪训明[1] 王顺德[1] 江行娟 杨庆云[1] 

机构地区：[1]复旦大学遗传所,杭州华东制药厂

出　　处：《中国抗生素杂志》1995年第5期333-339,共7页Chinese Journal of Antibiotics

摘　　要：从绛红色小单孢噬菌体Ｍｐｈ１ＤＮＡ中，用大肠杆菌启动子探测质粒ｐＧＡ４６克隆到在大肠杆菌中具有启动子功能的ＤＮＡ片段，筛选到的启动子功能片段具有抗四环素能力达１００μｇ／ｍｌ以上。对这些活性片段进行分子杂交，证明确实来自绛红色小单孢噬菌体Ｍｎｈ１。在此基础上，对其中一个２５０ｂｐ长的片段从克隆位点的一端进行了ＤＮＡ序列分析，其结构类似于大肠杆菌启动子。并进行了Ｇ＋Ｃ百分比的统计和限制性内切酶位点分析。Sau 3A restriction fragments from Micromonospora purpurea phage Mph1DNA have been cloned into promoter probe plasmid pGA46 which had been digested withBgl Ⅱ Transformants were selected on medium that allowed the selection of TCr promoterbearing plasmid.The origin of the cloned promoter was confirmed by means of Southern blothybridization,And,this fragment having E. coli promoter funcation has been sequenced byM13-dideoxynucleotide sequencing system. The result showed that molecular structure ofthis fragment is similar to that of E, coli,(G+C)%and site of varied restrictionendOnuclease were statistically calculated in this fragment in order to study its structure andfunction.

关 键 词：绛红色小单孢 噬菌体 克隆 DNA测序 抗生素 

分 类 号：TQ465.2[化学工程—制药化工]