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利用解淀粉芽孢杆菌的trpD+基因构建用于枯草芽孢杆菌宿主的启动子探针载体。

Construction of a promoter-probe vector for a Bacillus subtilis host by using the trpD+ gene of Bacillus amyloliquefaciens.

作者信息

Yoshimura K, Uemura J, Seki T, Oshima Y

出版信息

J Bacteriol. 1984 Sep;159(3):905-12. doi: 10.1128/jb.159.3.905-912.1984.

DOI:10.1128/jb.159.3.905-912.1984
PMID:6090398
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC215745/
Abstract

The trp gene cluster of Bacillus amyloliquefaciens was found to be structurally similar to that of the Enterobacteriaceae. The translation termination codon of the putative trpE gene and the initiation codon for the putative trpD gene overlap at the trpE-trpD junction, and a promoter for the putative trpC gene is suggested to exist. A promoter-probe vector of Bacillus subtilis, pFTB281, was constructed with a DNA fragment of B. amyloliquefaciens, complementing the trpC and trpD mutations of B. subtilis, a 42-base-pair DNA fragment of M13mp7, and the larger EcoRI-PvuII fragment of pUB110, which confers an autonomous replication function and the kanamycin-resistance phenotype to the chimeric plasmid. pFTB281 has BamHI, EcoRI, and SalI cloning sites in the 5'-upstream portion of the protein-coding region of the putative trpD gene, and the insertion of a certain DNA fragment at any of these sites allowed the plasmid to transform a trpD mutant of B. subtilis to the TrpD+ phenotype. DNA fragments showing the promoter function for the trpD gene were obtained from B. amyloliquefaciens and Saccharomyces cerevisiae chromosomes and rho 11 and lambda phage DNAs, but rarely from the DNAs of Escherichia coli and pBR322.

摘要

发现解淀粉芽孢杆菌的trp基因簇在结构上与肠杆菌科的相似。假定的trpE基因的翻译终止密码子与假定的trpD基因的起始密码子在trpE - trpD交界处重叠，并且推测存在假定的trpC基因的启动子。用解淀粉芽孢杆菌的一个DNA片段、补充枯草芽孢杆菌trpC和trpD突变的M13mp7的一个42碱基对DNA片段以及赋予嵌合质粒自主复制功能和卡那霉素抗性表型的pUB110的较大EcoRI - PvuII片段构建了枯草芽孢杆菌的一个启动子探针载体pFTB281。pFTB281在假定的trpD基因的蛋白质编码区5'上游部分有BamHI、EcoRI和SalI克隆位点，在这些位点中的任何一个插入特定DNA片段可使该质粒将枯草芽孢杆菌的trpD突变体转化为TrpD +表型。显示trpD基因启动子功能的DNA片段是从解淀粉芽孢杆菌和酿酒酵母染色体以及rho 11和λ噬菌体DNA中获得的，但很少从大肠杆菌DNA和pBR322中获得。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54b2/215745/ee05ff2be9e0/jbacter00232-0106-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54b2/215745/ee05ff2be9e0/jbacter00232-0106-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54b2/215745/ee05ff2be9e0/jbacter00232-0106-a.jpg

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本文引用的文献

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