Bondarenko V O, Kazennova E V, Bobkova M R, Prilipov A G, Bobkov A F
D. I. Ivanovsky Institute of Virology, Russian Academy of Medical Sciences, Moscow.
Mol Gen Mikrobiol Virusol. 2000(3):33-6.
Designing of recombinant plasmids pSB2 and pSB3 with the 932 bp HTLV-II env gene inserts encoding the full-length surface membrane glycoprotein gp46 is described. Vectors pGOmpF and pET32a expressing genes cloned under control of the late bacteriophage T7 promoter were used. Western blot analysis of cellular proteins derived from E. coli B834/pSB2 and E. coli B834/pSB3 revealed that 34 kD and 31 kD polypeptides corresponding to the full-length gp46 and its processed form without signal peptide were synthesized under control of these recombinant plasmids. Cytotoxic activity of the recombinant proteins towards bacterial cells was demonstrated. Both polypeptides specifically reacted to sera from humans infected with HTLV-II. High antigenic specificity of P34-HTLV-II proteins makes a promising candidate for diagnostic confirmation tests.
本文描述了重组质粒pSB2和pSB3的设计,其带有编码全长表面膜糖蛋白gp46的932 bp HTLV-II env基因插入片段。使用了在晚期噬菌体T7启动子控制下表达克隆基因的载体pGOmpF和pET32a。对源自大肠杆菌B834/pSB2和大肠杆菌B834/pSB3的细胞蛋白进行的蛋白质印迹分析表明,在这些重组质粒的控制下合成了对应于全长gp46及其无信号肽加工形式的34 kD和31 kD多肽。证明了重组蛋白对细菌细胞的细胞毒性活性。两种多肽都与感染HTLV-II的人类血清发生特异性反应。P34-HTLV-II蛋白的高抗原特异性使其成为诊断确认试验的有前景的候选物。