化脓性链球菌C型致热外毒素基因speC的克隆与特性分析
Cloning and characterization of the gene, speC, for pyrogenic exotoxin type C from Streptococcus pyogenes.
作者信息
Goshorn S C, Bohach G A, Schlievert P M
机构信息
University of Minnesota Medical School, Department of Microbiology, Minneapolis 55455.
出版信息
Mol Gen Genet. 1988 Apr;212(1):66-70. doi: 10.1007/BF00322445.
The structural gene of streptococcal pyrogenic exotoxin type C (SPE C) was cloned from the chromosome of Streptococcus pyogenes strain T18P into Escherichia coli using pBR328 as the vector plasmid. Subcloning enabled the localization of the gene (speC) to a 1.7 kb fragment. Partially purified E. coli-derived SPE C and purified streptococcal-derived SPE C, were shown to have the same molecular weight (23,800) and biological activities. A DNA probe, prepared from cloned speC, cross-hybridized with the structural genes of SPE A and SPE B indicating relatedness at the nucleotide level. The speC-derived probe also hybridized to a fragment of CS112 bacteriophage DNA containing the phage attachment site.
利用pBR328作为载体质粒，从化脓性链球菌T18P菌株的染色体中克隆出C型链球菌致热外毒素（SPE C）的结构基因，并将其导入大肠杆菌。亚克隆使得该基因（speC）定位到一个1.7 kb的片段上。结果显示，部分纯化的大肠杆菌衍生的SPE C和纯化的链球菌衍生的SPE C具有相同的分子量（23,800）和生物活性。从克隆的speC制备的DNA探针与SPE A和SPE B的结构基因发生交叉杂交，表明在核苷酸水平上具有相关性。speC衍生的探针还与含有噬菌体附着位点的CS112噬菌体DNA片段杂交。
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