Ermolaeva S A, Belyĭ Iu F, Tartakovskiĭ I S, Gintsburg A L, Prozorovskiĭ S V
Mol Gen Mikrobiol Virusol. 1994 Nov-Dec(6):3-8.
The gene for phosphatidylinositol-specific phospholipase C (PI-PLC) of Listeria monocytogenes has been cloned and shown to be expressed in Escherichia coli cells from own as well as from the lactose gene promoter. The recombinant plasmid has been constructed on the basis of pRIT2T vector and carries the hybrid gone. 3-end of which is a fragment of protein A gene of Staphylococcus aureus. 3-end is a gene for phospholipase plcA, both in the same reading frame. The resultant construction is shown to code in Escherichia coli cells for the hybrid recombinant protein A:Pl-PLC. Purified preparation of the hybrid protein and polyclonal rabbit antiserum to it were obtained. The obtained antiserum to the hybrid protein containing phospholipase as en C-end domain has been shown to react specifically to phospholipase in Escherichia coli recombinant strain harbouring the constructed recombinant plasmid as well as the one in the culture fluid of listeria.
单核细胞增生李斯特菌磷脂酰肌醇特异性磷脂酶C（PI-PLC）基因已被克隆，并显示在大肠杆菌细胞中可从自身以及乳糖基因启动子表达。重组质粒是在pRIT2T载体的基础上构建的，携带杂合基因。其3'端是金黄色葡萄球菌蛋白A基因的片段，3'端是磷脂酶plcA基因，两者处于同一阅读框。结果表明，该构建体在大肠杆菌细胞中编码杂合重组蛋白A:Pl-PLC。获得了杂合蛋白的纯化制剂及其多克隆兔抗血清。已证明，针对含有磷脂酶作为C端结构域的杂合蛋白所获得的抗血清，可与携带构建的重组质粒的大肠杆菌重组菌株以及李斯特菌培养液中的磷脂酶发生特异性反应。
