在表达3型荚膜多糖合成所需的肺炎球菌cap3A基因的大肠杆菌细胞提取物中UDP-葡萄糖脱氢酶活性的证明。
Demonstration of UDP-glucose dehydrogenase activity in cell extracts of Escherichia coli expressing the pneumococcal cap3A gene required for the synthesis of type 3 capsular polysaccharide.
作者信息
Arrecubieta C, García E, López R
机构信息
Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, Madrid, Spain.
出版信息
J Bacteriol. 1996 May;178(10):2971-4. doi: 10.1128/jb.178.10.2971-2974.1996.
Abstract
The gene cluster of Streptococcus pneumoniae coding for the type 3 capsular polysaccharide contains four genes (cap3ABCD). A DNA fragment containing the cap3A gene was amplified by PCR and cloned under the control of a T7 RNA polymerase-dependent promoter. Overexpression of this gene in Escherichia coli resulted both in a 47-kDa protein in the cytoplasm of isopropyl-beta-D-thiogalactopyranoside-induced bacteria and in high levels of UDP-glucose dehydrogenase activity. These data demonstrate, in a direct experimental way, that cap3A encodes the UDP-glucose dehydrogenase of pneumococcus type 3.
摘要
编码3型荚膜多糖的肺炎链球菌基因簇包含四个基因（cap3ABCD）。通过PCR扩增包含cap3A基因的DNA片段，并在T7 RNA聚合酶依赖性启动子的控制下进行克隆。该基因在大肠杆菌中的过表达导致异丙基-β-D-硫代半乳糖苷诱导的细菌细胞质中出现一种47 kDa的蛋白质，并产生高水平的UDP-葡萄糖脱氢酶活性。这些数据以直接实验的方式证明，cap3A编码3型肺炎球菌的UDP-葡萄糖脱氢酶。
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