基于转座子Tn5supF的反向遗传学方法，用于对克隆于λ噬菌体中的DNA的大肠杆菌进行突变分析。
Transposon Tn5supF-based reverse genetic method for mutational analysis of Escherichia coli with DNAs cloned in lambda phage.
作者信息
Phadnis S H, Kulakauskas S, Krishnan B R, Hiemstra J, Berg D E
机构信息
Department of Molecular Microbiology, Washington University Medical School, St. Louis, Missouri 63110.
出版信息
J Bacteriol. 1991 Jan;173(2):896-9. doi: 10.1128/jb.173.2.896-899.1991.
An efficient method for systematic mutational analysis of the Escherichia coli genome was developed. It entails Tn5supF transposition to lambda-E. coli hybrid phage clones (Kohara library) and then transduction of recipient cells to Sup+. Essential and nonessential genes are distinguished by the ability of insertion mutant phage to form haploid versus only heterozygous partial diploid bacterial recombinants.
开发了一种对大肠杆菌基因组进行系统突变分析的有效方法。该方法包括将Tn5supF转座到λ-大肠杆菌杂交噬菌体克隆（小原文库），然后将受体细胞转导为Sup+。通过插入突变噬菌体形成单倍体与仅形成杂合部分二倍体细菌重组体的能力来区分必需基因和非必需基因。
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