MAPPING OF A PLASMID, CODING FOR COLONIZATION FACTOR ANTIGEN-I AND HEAT-STABLE ENTERO-TOXIN PRODUCTION, ISOLATED FROM AN ENTERO-TOXIGENIC STRAIN OF ESCHERICHIA-COLI
MAPPING OF A PLASMID, CODING FOR COLONIZATION FACTOR ANTIGEN-I AND HEAT-STABLE ENTERO-TOXIN PRODUCTION, ISOLATED FROM AN ENTERO-TOXIGENIC STRAIN OF ESCHERICHIA-COLI
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DOI：
10.1128/jb.149.1.264-275.1982
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发表时间：
1982-01-01
影响因子：
3.2
通讯作者：
ROWE, B
中科院分区：
文献类型：
--
作者：
SMITH, HR;WILLSHAW, GA;ROWE, B
非自转移质粒 NTP113 编码产生定植因子抗原 I 和热稳定肠毒素。 NTP113，其MW为58倍。 106，用BamHI、EcoRI和HindIII以及这些限制性内切酶的组合进行消化，并通过琼脂糖凝胶电泳分析这些消化的产物。结果用于构建 NTP113 的部分限制性图谱。将编码氨苄青霉素、卡那霉素和四环素抗性的转座子插入NTP113中，通过携带插入突变体的菌株失去四环素或卡那霉素抗性来确定，获得了一系列缺失突变体。通过插入或缺失获得的许多质粒突变体不编码定植因子抗原I，但这些突变体中的大多数仍然编码热稳定肠毒素的产生。在限制性图谱上确定插入的转座子和缺失的位置。表达定植因子抗原 I 需要 NTP113 的两个区域，并且这两个位点由对应于大约 MW 的 DNA 长度分隔开。 25次。 106.
The non-autotransferring plasmid NTP113 codes for production of colonization factor antigen I and heat-stable enterotoxin. NTP113, which has a MW of 58 .times. 106, was digested with BamHI, EcoRI and HindIII and combinations of these restriction endonucleases, and the products of these digestions were analyzed by agarose gel electrophoresis. The results were used to construct a partial restriction map of NTP113. Transposons coding for resistance to ampicillin, kanamycin and tetracycline were inserted into NTP113, and a series of deletion mutants was obtained as determined by the loss of tetracycline or kanamycin resistance from strains carrying the insertion mutants. A number of plasmid mutants obtained by insertion or deletion did not code for colonization factor antigen I, but most of these mutants still coded for heat-stable enterotoxin production. The positions of the inserted transposons and of the deletions were determined on the restriction map. Two regions of NTP113 were required for the expression of colonization factor antigen I, and the 2 sites were separated by a length of DNA corresponding to a MW of .apprx. 25 .times. 106.