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
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
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DOI：
10.1128/jb.178.10.2971-2974.1996
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发表时间：
1996-05-01
影响因子：
3.2
通讯作者：
Lopez, R
中科院分区：
文献类型：
--
作者：
Arrecubieta, C;Garcia, E;Lopez, R
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 扩增含有 cap3A 基因的 DNA 片段，并将其克隆到 T7 RNA 聚合酶依赖性启动子的控制下。该基因在大肠杆菌中的过度表达导致异丙基-β-D-硫代半乳糖苷诱导的细菌的细胞质中产生 47 kDa 的蛋白质，并导致高水平的 UDP-葡萄糖脱氢酶活性。这些数据以直接实验方式证明 cap3A 编码 3 型肺炎球菌的 UDP-葡萄糖脱氢酶。
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.