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首页> 外文期刊>Journal of bacteriology >Overproduction and identification of the ftsQ gene product, an essential cell division protein in Escherichia coli K-12.
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Overproduction and identification of the ftsQ gene product, an essential cell division protein in Escherichia coli K-12.

机译：ftsQ基因产物的过量生产和鉴定，ftsQ基因产物是大肠杆菌K-12中必需的细胞分裂蛋白。

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ftsQ is an essential cell division gene in Escherichia coli. The ftsQ gene has been sequenced, and a presumptive open reading frame has been identified; however, no protein product has been observed (A.C. Robinson, D.J. Kenan, G.F. Hatfull, N.F. Sullivan, R. Spiegelberg, and W.D. Donachie, J. Bacteriol. 160:546-555, 1984, and Q.M. Yi, S. Rockenbach, J.E. Ward, Jr., and J. Lutkenhaus, J. Mol. Biol. 184:399-412, 1985). The ftsQ gene was isolated on a 970-base-pair EcoRI-PvuII fragment of the E. coli chromosome and used to construct a trp-lac (Ptac) transcriptional fusion in plasmid pKK223-3. The fused construct (pDSC78) complemented an ftsQ1(Ts) mutant strain in trans, restoring growth at 42 degrees C on low-salt medium. An ftsQ1(Ts) mutant strain transformed with pDSC78 appeared normal upon microscopic examination, with no indication of filamentation. The ftsQ gene product was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional gel electrophoresis of radiolabeled, isopropyl-beta-D-thiogalactopyranoside-induced maxicell and normal cell extracts. As predicted from the nucleotide sequence, the 970-base-pair EcoRI-PvuII fragment encoded a polypeptide of approximately 31,400 daltons. Analysis of the data obtained from pulse-chase experiments in maxicells and normal cells suggests that the FtsQ protein is stable. Most of the radiolabeled FtsQ protein from maxicells was found in the inner membrane. On the basis of available information, the prior inability to detect FtsQ can be attributed to low levels of transcription or translation rather than to proteolysis.
机译：ftsQ是大肠杆菌中必不可少的细胞分裂基因。已对ftsQ基因进行了测序，并鉴定了一个假定的开放阅读框;但是，未观察到蛋白质产物（1984年AC Robinson，DJ Kenan，GF Hatfull，NF Sullivan，R.Spiegelberg和WD Donachie，J. Bacteriol。160：546-555，以及QM Yi，S. Rockenbach，JE Ward，Jr。和J.Lutkenhaus，J.Mol.Biol.184：399-412，1985）。在大肠杆菌染色体的970个碱基对的EcoRI-PvuII片段上分离了ftsQ基因，并用于在质粒pKK223-3中构建trp-lac（Ptac）转录融合体。融合构建体（pDSC78）反式补充了ftsQ1（Ts）突变株，在低盐培养基上恢复了42摄氏度的生长。经pDSC78转化的ftsQ1（Ts）突变菌株在显微镜检查下显示正常，没有丝状迹象。 ftsQ基因产物通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和放射性标记的异丙基-β-D-硫代吡喃半乳糖苷诱导的Maxicell和正常细胞提取物的二维凝胶电泳进行鉴定。如从核苷酸序列所预测的，970个碱基对的EcoRI-PvuII片段编码的多肽约为31,400道尔顿。从脉搏追踪实验在下颌和正常细胞中获得的数据分析表明，FtsQ蛋白是稳定的。大部分来自maxicells的放射性标记的FtsQ蛋白都在内膜中发现。根据可用信息，先前无法检测到FtsQ的原因可归因于转录或翻译水平较低，而不是蛋白水解。

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