Cherepanov P A, Mikhaĭlova T G, Karimova G A, Zakharova N M, Ershov Iu V, Volkovoĭ K I
Mol Gen Mikrobiol Virusol. 1991 Dec(12):19-26.
The genetical libraries of the pFra plasmid of Yersinia pestis genes were obtained by insertion into the PstI, SalGI, EcoRI, XhoI restriction sites of the cosmid vector pHC79. The immunochemical analysis of the recombinant clones has revealed the clones synthesizing the antigen Fl (fraction I) and mouse toxin (Ymt--Yersinia pestis murine toxin). The restriction analysis of the plasmids from antigen synthesizing clones has permitted to construct the detailed physical map of the fra-ymt region of the pFra plasmid the size of 22 kb. The recombinant F1 positive clones of Escherichia coli are able to form at 37 degrees C the capsule-like structure peculiar for Yersinia pestis. The antigen F1 and the mouse toxin were isolated, purified and characterized. The antigen F1 is an 1-2 Md polymer containing a 16 kDa protein subunit. The mouse toxin a 240 kDa protein consisting of 61 kDa subunits. The nucleotide sequence of ymt gene has been defined.
通过插入黏粒载体pHC79的PstI、SalGI、EcoRI、XhoI限制性酶切位点，获得了鼠疫耶尔森氏菌基因的pFra质粒基因文库。对重组克隆的免疫化学分析揭示了合成抗原F1（组分I）和小鼠毒素（Ymt——鼠疫耶尔森氏菌小鼠毒素）的克隆。对抗原合成克隆的质粒进行限制性分析，得以构建出大小为22 kb的pFra质粒fra - ymt区域的详细物理图谱。大肠杆菌的重组F1阳性克隆在37℃时能够形成鼠疫耶尔森氏菌特有的类荚膜结构。抗原F1和小鼠毒素被分离、纯化并进行了特性鉴定。抗原F1是一种1 - 2 Md的聚合物，含有一个16 kDa的蛋白质亚基。小鼠毒素是一种由61 kDa亚基组成的240 kDa蛋白质。已确定了ymt基因的核苷酸序列。
