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大肠杆菌35 kDa可溶性溶菌转糖基酶编码基因的克隆与可控过表达。

Cloning and controlled overexpression of the gene encoding the 35 kDa soluble lytic transglycosylase from Escherichia coli.

作者信息

Dijkstra A J, Hermann F, Keck W

机构信息

Pharma Research Department, Hoffmann-La Roche Ltd., Basel, Switzerland.

出版信息

FEBS Lett. 1995 Jun 12;366(2-3):115-8. doi: 10.1016/0014-5793(95)00505-4.

Abstract

The lytic transglycosylases of Escherichia coli are involved in peptidoglycan metabolism and resemble the lysozymes not only in activity, but in the case of the 70 kDa soluble lytic transglycosylase (Slt70), also structurally. Here we report the cloning of the gene that encodes the 35 kDa soluble lytic transglycosylase (Slt35) of E. coli. Based on the sequence of the full-length gene, Slt35 is very likely to be a proteolytically truncated form of a slightly larger protein. The homology between Slt35 and Slt70, albeit poor, indicates that the active site architecture of both proteins may be alike. Using the T-7 promoter system, Slt35 was overproduced in large quantities and purified to homogeneity for crystallographic purposes.

摘要

大肠杆菌的溶菌转糖基酶参与肽聚糖代谢,不仅在活性上与溶菌酶相似,而且就70 kDa可溶性溶菌转糖基酶(Slt70)而言,在结构上也与溶菌酶相似。在此,我们报道了编码大肠杆菌35 kDa可溶性溶菌转糖基酶(Slt35)的基因的克隆。根据全长基因的序列,Slt35很可能是一种略大蛋白质的蛋白水解截短形式。Slt35和Slt70之间的同源性虽然很低,但表明这两种蛋白质的活性位点结构可能相似。利用T-7启动子系统,大量过量表达Slt35并将其纯化至同质,以用于晶体学研究。

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