Lindsay D A, Vonderfecht S S, Willoughby R, Betenbaugh M J, Eiden J J
Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218.
Virology. 1993 Jun;194(2):724-33. doi: 10.1006/viro.1993.1313.
The polypeptide encoded by gene 3 of the IDIR strain of group B rotavirus (GBR) was synthesized by means of a baculovirus expression system. Immunoblot analysis identified the IDIR virus gene 3 product as analogous to the outer capsid protein, VP4, encoded by gene 4 of group A rotaviruses (GAR). This coding assignment had previously been difficult to establish by sequence comparisons, since IDIR virus gene 3 shared < 20% identical amino acid sequences with any GAR protein. Trypsin digestion of the gene 3 protein resulted in the appearance of a product indistinguishable in size from the VP5* outer capsid protein of IDIR agent virions. The recombinant IDIR virus VP4 maintained at least some antigenic epitopes of the native virion protein, as evidenced by reactivity with convalescent antibody obtained following infection of infant rat pups with the IDIR agent. Reactivity was also demonstrated with antisera directed against bovine and porcine isolates of GBR as well as with ADRV, a human strain of GBR.
通过杆状病毒表达系统合成了B组轮状病毒(GBR)IDIR株基因3编码的多肽。免疫印迹分析确定IDIR病毒基因3产物类似于A组轮状病毒(GAR)基因4编码的外衣壳蛋白VP4。由于IDIR病毒基因3与任何GAR蛋白的氨基酸序列同源性小于20%,以前很难通过序列比较确定这种编码关系。基因3蛋白经胰蛋白酶消化后,产生了一种大小与IDIR病原体病毒粒子的VP5*外衣壳蛋白无法区分的产物。重组IDIR病毒VP4保留了天然病毒粒子蛋白的至少一些抗原表位,这一点可通过用IDIR病原体感染新生幼鼠后获得的恢复期抗体的反应性得到证明。针对GBR牛和猪分离株的抗血清以及针对GBR人毒株ADRV的抗血清也显示出反应性。