马链球菌兽疫亚种H46A中脱氧核糖核酸酶（链道酶）编码基因在大肠杆菌、枯草芽孢杆菌和乳酸乳球菌中的分离、测序及表达

Isolation, sequence and expression in Escherichia coli, Bacillus subtilis and Lactococcus lactis of the DNase (streptodornase)-encoding gene from Streptococcus equisimilis H46A.

作者信息

Wolinowska R, Cegłowski P, Kok J, Venema G

机构信息

Department of Pharmaceutical Microbiology, Medical Academy, Warsaw, Poland.

出版信息

Gene. 1991 Sep 30;106(1):115-9. doi: 10.1016/0378-1119(91)90574-u.

DOI:10.1016/0378-1119(91)90574-u

PMID:1937032

Abstract

A partial library of BclI-generated chromosomal DNA fragments from Streptococcus equisimilis H64A (Lancefield Group C) was constructed in Escherichia coli. Clones displaying either streptokinase or deoxyribonuclease (streptodornase; SDC) activities were isolated. The gene (sdc) expressing the SDC activity was allocated on the 1.1-kb AccI DNA subfragment. Sequence analysis of this DNA fragment revealed the presence of one open reading frame, which could encode a protein of 36.8 kDa. The N-terminal portion of the deduced protein exhibited features characteristic of prokaryotic signal peptides. The sdc gene was expressed in E. coli, Bacillus subtilis and Lactococcus lactis. As observed for S. equisimilis, in the heterologous Gram + hosts, at least part of the SDC protein was secreted into the medium.

摘要

构建了来自马链球菌兽疫亚种H64A（兰斯菲尔德C组）的由BclI产生的染色体DNA片段的部分文库，该文库存在于大肠杆菌中。分离出了显示链激酶或脱氧核糖核酸酶（链道酶；SDC）活性的克隆。表达SDC活性的基因（sdc）定位于1.1 kb的AccI DNA亚片段上。对该DNA片段的序列分析揭示存在一个开放阅读框，其可编码一种36.8 kDa的蛋白质。推导蛋白质的N端部分具有原核信号肽的特征。sdc基因在大肠杆菌、枯草芽孢杆菌和乳酸乳球菌中表达。正如在马链球菌兽疫亚种中观察到的那样，在异源革兰氏阳性宿主中，至少部分SDC蛋白分泌到了培养基中。