MOLECULAR-CLONING AND EXPRESSION OF HIPA, A GENE OF ESCHERICHIA-COLI K-12 THAT AFFECTS FREQUENCY OF PERSISTENCE AFTER INHIBITION OF MUREIN SYNTHESIS
MOLECULAR-CLONING AND EXPRESSION OF HIPA, A GENE OF ESCHERICHIA-COLI K-12 THAT AFFECTS FREQUENCY OF PERSISTENCE AFTER INHIBITION OF MUREIN SYNTHESIS
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DOI：
10.1128/jb.166.2.399-403.1986
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发表时间：
1986-05-01
影响因子：
3.2
通讯作者：
BRODERICK, SH
中科院分区：
文献类型：
--
作者：
MOYED, HS;BRODERICK, SH
大肠杆菌染色体上33.8 min处的hipA基因控制胞壁素合成抑制后的持续频率;对于携带hipA+的菌株，频率为10-6，对于hipA-菌株，频率为10-2。通过在33.9 min时选择卡那霉素抗性决定簇克隆了hip+。在recA+和recA-背景中，hipA+均比hipA-占优势。根据质粒与hipA-菌株互补的能力，含有hipA+的最小DNA插入片段为1，885个碱基对。当载体的克隆位点远离强启动子时，可以获得两个方向的hipA+;两个方向都与hipA-互补，并且都编码50，000 Mr的独特肽。从质粒编码的肽的模式中推断出可能的转录方向，其中插入物和相邻载体序列的任一端已被删除。这一信息以及当克隆位点靠近强启动子时hipA+仅恢复一个方向表明，该基因的高水平表达不被E.杆菌
The hipA gene at 33.8 min on the Escherichia coli chromosome controls the frequency of persistence upon inhibition of murein synthesis; for strains bearing hipA+ the frequency is 10-6, and for hipA- strains the frequency is 10-2. hip+ has been cloned by selection for a kanamycin resistance determinant at 33.9 min. hipA+ is dominant over hipA- in both recA+ and recA- backgrounds. The smallest DNA insert which contains hipA+, as determined by the ability of the plasmids to complement hipA- strains, is 1,885 base pairs. Both orientations of hipA+ are obtained when the cloning site of vector is remote from strong promoters; both orientations complement hipA-, and both encode a unique peptide of 50,000 Mr. The probable direction of transcription has been deduced from the pattern of peptides encoded by plasmids from which either end of the insert and adjacent vector sequences have been deleted. This information and the recovery of only one orientation of hipA+ when the cloning site is close to a strong promoter suggest that a high level of expression of the gene is not tolerated by E. coli.