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首页> 外文期刊>Infection and immunity >The gene coding for the 190,000-dalton iron-regulated protein of Yersinia species is present only in the highly pathogenic strains.
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The gene coding for the 190,000-dalton iron-regulated protein of Yersinia species is present only in the highly pathogenic strains.

机译：编码190,000道尔顿铁调控的耶尔森氏菌种的基因仅存在于高致病性菌株中。

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摘要

A genomic library containing DNA fragments of 0.5 to 2 kilobase pairs in length from Yersinia enterocolitica serovar O:8 was constructed in a bacteriophage lambda gt11 expression vector. Mouse antibodies specific for the iron-regulated high-molecular-weight proteins (HMWPs) were used to screen the library. Two positive clones of 1 and 0.5 kilobase pairs, designated A13 and D7, respectively, were detected and isolated. They coded for beta-galactosidase fusion proteins of 151,000 and 138,000 daltons (Da). Antibodies affinity purified on the two recombinant lambda gt11 vectors specifically recognized the smaller HMWP (190,000 Da) and not the larger (240,000 Da). The two cloned DNA fragments were used to construct recombinant amplification plasmid pUC13 and to obtain large amounts of purified A13 and D7 inserts. Southern hybridizations performed with the inserts used as probes revealed that: (i) the two cloned DNA fragments overlap; (ii) only one gene hybridizes with the A13 and D7 inserts; (iii) the gene coding for the HMWP is conserved among all highly pathogenic Yersinia species studied; (iv) this gene is missing in the low-virulence and nonvirulent strains; and (v) transcription of the HMWP gene is induced by iron starvation.
机译：在λ噬菌体gt11表达载体中构建了一个基因组文库，该文库包含来自肠小肠结肠炎耶尔森氏菌O：8的0.5至2个碱基对的DNA片段。对铁调节的高分子量蛋白（HMWP）具有特异性的小鼠抗体用于筛选文库。检测并分离出两个分别为1和0.5碱基对的阳性克隆，分别命名为A13和D7。他们编码了151,000和138,000道尔顿（Da）的β-半乳糖苷酶融合蛋白。在两个重组λgt11载体上纯化的亲和力抗体特异性识别较小的HMWP（190,000 Da），而不识别较大的HMWP（240,000 Da）。这两个克隆的DNA片段用于构建重组扩增质粒pUC13，并获得大量纯化的A13和D7插入片段。用插入物作为探针进行的Southern杂交显示：（i）两个克隆的DNA片段重叠; （ii）只有一个基因与A13和D7插入片段杂交; （iii）在所有研究的高致病性耶尔森氏菌中，编码HMWP的基因是保守的; （iv）在低毒力和非毒力菌株中该基因缺失; （v）铁饥饿导致HMWP基因的转录。

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