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植物乳杆菌镉和锰吸收基因的克隆、表达及特性分析

Cloning, expression, and characterization of cadmium and manganese uptake genes from Lactobacillus plantarum.

作者信息

Hao Z, Chen S, Wilson D B

机构信息

Institute for Comparative and Environmental Toxicology, Cornell University, Ithaca, New York 14853, USA.

出版信息

Appl Environ Microbiol. 1999 Nov;65(11):4746-52. doi: 10.1128/AEM.65.11.4746-4752.1999.

Abstract

An Mn(2+) and Cd(2+) uptake gene, mntA, was cloned from Lactobacillus plantarum ATCC 14917 into Escherichia coli. Its expression conferred on E. coli cells increased Cd(2+) sensitivity as well as energy-dependent Cd(2+) uptake activity. Both transcription and translation of mntA were induced by Mn(2+) starvation in L. plantarum, as indicated by reverse transcriptase PCR and immunoblotting. Two Cd(2+) uptake systems have been identified in L. plantarum: one is a high-affinity Mn(2+) and Cd(2+) uptake system that is expressed in Mn(2+)-starved cells, and the other is a nonsaturable Cd(2+) uptake system that is expressed in Cd(2+)-sufficient cells (Z. Hao, H. R. Reiske, and D. B. Wilson, Appl. Environ. Microbiol. 65:592-99, 1999). MntA was not detected in an Mn(2+)-dependent mutant of L. plantarum which had lost high-affinity Mn(2+) and Cd(2+) uptake activity. The results suggest that mntA is the gene encoding the high-affinity Mn(2+) and Cd(2+) transporter. On the basis of its predicted amino acid sequence, MntA belongs to the family of P-type cation-translocating ATPases. The topology and potential Mn(2+)- and Cd(2+)-binding sites of MntA are discussed. A second clone containing a low-affinity Cd(2+) transport system was also isolated.

摘要

从植物乳杆菌ATCC 14917中克隆出一个锰离子(Mn(2+))和镉离子(Cd(2+))摄取基因mntA,并将其导入大肠杆菌。该基因的表达使大肠杆菌细胞对Cd(2+)的敏感性增加,同时赋予其能量依赖性的Cd(2+)摄取活性。逆转录聚合酶链反应(RT-PCR)和免疫印迹结果表明,在植物乳杆菌中,mntA的转录和翻译均受Mn(2+)饥饿诱导。在植物乳杆菌中已鉴定出两种Cd(2+)摄取系统:一种是在Mn(2+)饥饿细胞中表达的高亲和力Mn(2+)和Cd(2+)摄取系统,另一种是在Cd(2+)充足细胞中表达的非饱和性Cd(2+)摄取系统(Z. Hao、H. R. Reiske和D. B. Wilson,《应用与环境微生物学》65:592 - 99,1999年)。在植物乳杆菌的一个Mn(2+)依赖性突变体中未检测到MntA,该突变体已丧失高亲和力Mn(2+)和Cd(2+)摄取活性。结果表明,mntA是编码高亲和力Mn(2+)和Cd(2+)转运蛋白的基因。根据其预测的氨基酸序列,MntA属于P型阳离子转运ATP酶家族。文中还讨论了MntA的拓扑结构以及潜在的Mn(2+)和Cd(2+)结合位点。此外,还分离出了一个包含低亲和力Cd(2+)转运系统的第二个克隆。

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