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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Construction and characterization of Streptococcus suis-Escherichia coli shuttle cloning vectors.
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Construction and characterization of Streptococcus suis-Escherichia coli shuttle cloning vectors.

机译：猪链球菌-大肠杆菌穿梭克隆载体的构建和鉴定。

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pSSU1, a native plasmid of Streptococcus suis DAT1, was used to construct pSET-series shuttle vectors. In addition to the replication function of pSSU1, these vectors contain the multiple cloning sites and lacZ' gene from pUC19, which means that X-gal screening can be used to select recombinants in Escherichia coli. pSET1, pSET2, and pSET3 carry cat, spc, and both of these genes, respectively, as selectable markers. These vectors could be introduced into S. suis, E. coli, Salmonella typhimurium, S. pneumoniae, and S. equi ssp. equi by electrotransformation. The recA gene was cloned from S. suis and sequenced, and this information was used in the construction of a recA mutant of S. suis. Transformation frequencies and/or plasmid stability of all pSET vectors tested were decreased in both S. suis and E. coli recA mutants compared with the parental strains. These results suggested that functional RecA protein improved the maintenance of pSET vectors in both S. suis and E. coli. Moreover, cloning of the functional S. suis recA gene into pSET2 and complementation analysis of the recA mutant were successful in S. suis but not in E. coli. These results showed that pSET vectors are useful tools for cloning and analyzing S. suis genes in S. suis strains directly. Copyright 2001 Academic Press.
机译：猪链球菌DAT1的天然质粒pSSU1用于构建pSET系列穿梭载体。这些载体除了具有pSSU1的复制功能外，还包含多个克隆位点和来自pUC19的lacZ'基因，这意味着X-gal筛选可用于选择大肠杆菌中的重组体。 pSET1，pSET2和pSET3分别携带cat，spc和这两个基因作为选择标记。这些载体可被引入猪链球菌，大肠杆菌，鼠伤寒沙门氏菌，肺炎链球菌和马链球菌。通过电转化产生的当量。从猪链球菌克隆recA基因并测序，并将该信息用于猪链球菌recA突变体的构建。与亲代菌株相比，猪链球菌和大肠杆菌recA突变体中所有测试的pSET载体的转化频率和/或质粒稳定性均降低。这些结果表明，功能性RecA蛋白改善了猪链球菌和大肠杆菌中pSET载体的维持。此外，将功能性猪链球菌recA基因克隆到pSET2中和recA突变体的互补分析在猪链球菌中成功，但在大肠杆菌中却没有。这些结果表明，pSET载体是直接克隆和分析猪链球菌菌株中猪链球菌基因的有用工具。版权所有2001学术出版社。

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