Lindsay D A, Vonderfecht S L, Eiden J J
Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218.
Virology. 1994 Feb 15;199(1):141-50. doi: 10.1006/viro.1994.1106.
The second largest genomic segment of the IDIR strain (infectious diarrhea of infant rats) of group B rotavirus (GBR) was completely sequenced, cloned, and expressed in insect cells, and its gene coding assignment was determined. The sequence of IDIR virus gene 2 (IDIRg2) contained 2847 bp with a single, long open reading frame that encoded a deduced polypeptide of 934 amino acids (M(r) 106 kDa, pI 5.325). BestFit homology indicated that the predicted amino acid sequence of IDIRg2 shared 46.5% similar and 19.8% identical sequences with VP2 of the SA11 strain of group A rotavirus. The polypeptide product encoded by this gene was synthesized in insect cells by means of a baculovirus expression vector and employed to elucidate the corresponding gene to protein coding assignment. Recombinant IDIRg2 product maintained virion antigenic epitopes as evidenced by reactivity with convalescent antisera from infant rat pups infected with the IDIR agent. Reactivity with antisera from heterologous human and porcine GBR strains was also observed. Antibody directed against recombinant IDIRg2 product specifically reacted with VP2 of IDIR virus and a human strain of GBR (ADRV) obtained from fecal specimens. These experiments identified the IDIRg2 product as the VP2 core protein of the IDIR virus strain of GBR.
对B组轮状病毒（GBR）的IDIR株（幼鼠感染性腹泻）的第二大基因组片段进行了全序列测定、克隆，并在昆虫细胞中表达，同时确定了其基因编码归属。IDIR病毒基因2（IDIRg2）的序列包含2847 bp，有一个单一的长开放阅读框，编码一个推导的934个氨基酸的多肽（M(r) 106 kDa，pI 5.325）。BestFit同源性分析表明，IDIRg2预测的氨基酸序列与A组轮状病毒SA11株的VP2序列有46.5%的相似性和19.8%的同一性。该基因编码的多肽产物通过杆状病毒表达载体在昆虫细胞中合成，并用于阐明相应的基因到蛋白质的编码归属。重组IDIRg2产物保留了病毒粒子的抗原表位，这可通过与感染IDIR病原体的幼鼠恢复期抗血清的反应性得到证明。还观察到与来自异源人类和猪GBR株的抗血清的反应性。针对重组IDIRg2产物的抗体与从粪便标本中获得的IDIR病毒和一种人类GBR株（ADRV）的VP2发生特异性反应。这些实验确定IDIRg2产物为GBR的IDIR病毒株的VP2核心蛋白。
