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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >DEVELOPMENT OF A RHODOCOCCUS EQUI ESCHERICHIA COLI PLASMID SHUTTLE VECTOR
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DEVELOPMENT OF A RHODOCOCCUS EQUI ESCHERICHIA COLI PLASMID SHUTTLE VECTOR

机译：马氏红球菌大肠杆菌溶质穿梭载体的开发

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Isolates of Rhodococcus equi from pneumonic foals possess an 85- or 90-kb virulence-associated plasmid. A prominent, thermoregulated surface antigen, VapA, encoded by these plasmids is thought to be important in virulence. A 1.35-kb fragment containing the origin of replication of R. equi strain 103 virulence-associated plasmid (pOTS) was identified, sequenced, and its location identified. A simple R. equi-Escherichia coli shuttle plasmid (pRE-1) derived from the E. coli plasmid pACYC177 and the pOTS ori was developed. The plasmid transformed readily and was stable in either host and expressed kanamycin resistance but not beta-lactamase in R. equi. An improved 5.9-kb vector, pRE-7, was developed from pRE-1 and pBluescript. Subcloning of vapA into the multiple cloning site of the beta-galactosidase gene of pRE-7 resulted in weak expression of the gene both in E. coli and R. equi. The shuttle vector may be useful in examining regulation of virulence gene expression in R. equi. (C) 1997 Academic Press. [References: 31]
机译：从肺炎小马驹分离出的马红球菌具有85或90 kb与毒力相关的质粒。由这些质粒编码的突出的，受温度调节的表面抗原VapA被认为对毒力很重要。一个1.35 kb的片段，包含复制的马R. equi菌株103毒力相关质粒（pOTS）的起点，进行了测序，并确定了其位置。开发了一种简单的源自大肠杆菌质粒pACYC177和pOTS ori的大肠杆菌大肠杆菌穿梭质粒（pRE-1）。该质粒易于转化，并且在任一宿主中均稳定，并在卡那霉素中表达卡那霉素抗性，但不表达β-内酰胺酶。从pRE-1和pBluescript开发出改进的5.9-kb载体pRE-7。将vapA亚克隆到pRE-7的β-半乳糖苷酶基因的多克隆位点中，导致该基因在大肠杆菌和马蝇中均表达较弱。穿梭载体可用于检查对马鞭毛虫中毒力基因表达的调节。 （C）1997学术出版社。 [参考：31]

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