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首页> 外文期刊>Infection and immunity >Cloning and molecular characterization of the B subunit of Escherichia coli heat-labile enterotoxin.
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Cloning and molecular characterization of the B subunit of Escherichia coli heat-labile enterotoxin.

机译：大肠杆菌热不稳定肠毒素B亚基的克隆和分子鉴定。

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摘要

We have constructed a plasmid containing the gene for production of the B subunit of the heat-labile enterotoxin (LT-B) from a human isolate of Escherichia coli, strain H10407. The 0.8-kilobase gene fragment encoding synthesis of LT-B was cloned onto plasmid pBR322 after sequential digestion of the enterotoxin plasmid of strain H10407 with restriction endonucleases PstI and HindIII. LT-B was isolated by agarose affinity chromatography from cell lysates of recombinant clones expressing the B subunit. The B subunit was isolated in its oligomeric form, was structurally identical to native B subunit when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, dissociated to monomeric B in the presence of 5 M guanidine, was immunologically identical to native B subunit in an enzyme-linked immunosorbent assay, and contained no demonstrable A subunit in any of the assays.
机译：我们已经构建了一个质粒，该质粒包含用于从大肠杆菌的人类分离株H10407生产不耐热肠毒素（LT-B）B亚基的基因。在用限制性核酸内切酶PstI和HindIII顺序消化菌株H10407的肠毒素质粒后，将编码LT-B合成的0.8-碱基碱基的基因片段克隆到质粒pBR322上。通过琼脂糖亲和层析从表达B亚基的重组克隆的细胞裂解物中分离出LT-B。 B亚基以寡聚体形式分离，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检查时与天然B亚基结构相同，在5 M胍存在下与单体B解离，在酶学上与天然B亚基免疫学相同联免疫吸附测定，并且在任何测定中均不含可证明的A亚基。

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