Nucleotide sequence and expression in Escherichia coli of the Pseudomonas aeruginosa lasA gene.
Nucleotide sequence and expression in Escherichia coli of the Pseudomonas aeruginosa lasA gene.
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铜绿假单胞菌 lasA 基因的核苷酸序列和在大肠杆菌中的表达。
DOI：
10.1128/jb.170.6.2784-2789.1988
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发表时间：
1988
影响因子：
3.2
通讯作者：
Iglewski,BH
中科院分区：
文献类型：
--
作者：
Schad,PA;Iglewski,BH
铜绿假单胞菌PAO-E64是一种突变株，它能产生亲本水平的弹性蛋白酶抗原，但在37℃下没有弹性裂解活性。PAO-E64中的损伤(LasA1)不是铜绿假单胞菌弹性蛋白酶结构基因的突变(P.A.Schad，R.A.Bever，T.I.Nicas，F.Leduce，L.F.Hanne和B.H.Iglewski，J.细菌素)。169：2691-2696,1987)。对补充LasA1损伤的1.7kb的DNA片段进行了测序。对DNA序列的计算机分析显示，它含有一个开放阅读框，编码一个41,111-道尔顿蛋白。LASA基因在可诱导的PT-7启动子下表达，并在大肠杆菌裂解产物中检测到40,000道尔顿蛋白。LASA蛋白定位于大肠杆菌外膜部分。这种在大肠杆菌中产生的LASA蛋白激活了铜绿假单胞菌突变株PAO-E64产生的胞外弹性蛋白酶。
Pseudomonas aeruginosa PAO-E64 is a mutant which produces parental levels of elastase antigen but has no elastolytic activity at 37 degrees C. The lesion (lasA1) in PAO-E64 is not a mutation in the structural gene for P. aeruginosa elastase (P.A. Schad, R.A. Bever, T.I. Nicas, F. Leduce, L.F. Hanne, and B.H. Iglewski, J. Bacteriol. 169: 2691-2696, 1987). A 1.7-kilobase segment of DNA that complements the lasA1 lesion was sequenced. Computer analysis of the DNA sequence showed that it contained an open reading frame which encoded a 41,111-dalton protein. The lasA gene was expressed under an inducible PT-7 promoter, and a 40,000-dalton protein was detected in Escherichia coli lysates. The lasA protein was localized in the outer membrane fraction of E. coli. This lasA protein produced in E. coli activated the extracellular elastase produced by the P. aeruginosa mutant, PAO-E64.