克隆促进基因在枯草芽孢杆菌中表达的限制性片段。
Cloning restriction fragments that promote expression of a gene in Bacillus subtilis.
作者信息
Williams D M, Duvall E J, Lovett P S
出版信息
J Bacteriol. 1981 Jun;146(3):1162-5. doi: 10.1128/jb.146.3.1162-1165.1981.
Abstract
Plasmid pPL603 (3.1 megadaltons) specifies neomycin resistance in Bacillus subtilis and contains a structural gene for chloramphenicol acetyltransferase. Cells harboring the plasmid cannot grow on solid media containing 10 microgram of chloramphenicol per ml. Cloning EcoRI (or EcoRI)-generated fragments of deoxyribonucleic acid from several sources into the single EcoRI site in plasmid pPL603, with subsequent selection of transformants of media containing 10 micrograms of chloramphenicol per ml, permits the identification of restriction fragments that promote expression of the chloramphenicol acetyltransferase gene.
摘要
质粒pPL603（3.1兆道尔顿）赋予枯草芽孢杆菌新霉素抗性，并含有氯霉素乙酰转移酶的结构基因。携带该质粒的细胞不能在每毫升含10微克氯霉素的固体培养基上生长。将来自多个来源的经EcoRI（或EcoRI）酶切产生的脱氧核糖核酸片段克隆到质粒pPL603的单一EcoRI位点，随后在每毫升含10微克氯霉素的培养基上筛选转化体，这样就能鉴定出促进氯霉素乙酰转移酶基因表达的限制性片段。
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