Jiang B M, Qian Y, Tsunemitsu H, Green K Y, Saif L J
Food Animal Health Research Program, Ohio State University, Wooster, 44691.
Virology. 1991 Sep;184(1):433-6. doi: 10.1016/0042-6822(91)90864-8.
The genetic diversity of gene 8 (encoding the outer capsid glycoprotein VP7) among group (Gp) C rotaviruses was examined by Northern and dot blot hybridization. A cDNA clone of the porcine Gp C Cowden strain gene 8 was labeled with 32P by nick translation and used as a probe. The gene 8 probe hybridized with the corresponding gene of one human (88-196) and four porcine (Cowden, NB, WH, and Wi) strains of Gp C rotaviruses under both moderate (50% formamide, 5X SSC, and 42 degrees) and high (50% formamide, 5X SSC, and 52 degrees) stringency conditions. However, under high stringency conditions little or no hybridization was detected with the corresponding gene of one bovine (Shintoku) and three other porcine (Ah, HF, and KH) strains of Gp C rotaviruses. In control experiments, the Cowden gene 8 probe did not hybridize with Gp A (Gottfried strain) or Gp B (Ohio strain) rotaviruses. These data demonstrate that the Cowden gene 8 probe is Gp C rotavirus-specific and that genetic diversity exists among Gp C rotaviruses in the gene encoding the outer capsid glycoprotein VP7. Our gene 8 probe may be useful in hybridization assays for serotyping Gp C rotaviruses, analogous to the use of VP7 probes for serotyping Gp A rotaviruses. However, final confirmation of our genetic approach to serotype Gp C rotaviruses awaits the serologic analysis of these viruses.
通过Northern杂交和点杂交检测了C组轮状病毒中基因8（编码外衣壳糖蛋白VP7）的遗传多样性。用缺口平移法将猪C组考登株基因8的cDNA克隆用32P标记，并用作探针。在中等严谨条件（50%甲酰胺、5×SSC和42℃）和高严谨条件（50%甲酰胺、5×SSC和52℃）下，基因8探针与一株人C组轮状病毒（88 - 196）及四株猪C组轮状病毒（考登、NB、WH和Wi）的相应基因发生杂交。然而，在高严谨条件下，未检测到与一株牛C组轮状病毒（新德株）及其他三株猪C组轮状病毒（Ah、HF和KH）的相应基因发生杂交。在对照实验中，考登基因8探针未与A组（戈特弗里德株）或B组（俄亥俄株）轮状病毒杂交。这些数据表明考登基因8探针是C组轮状病毒特异性的，并且在编码外衣壳糖蛋白VP7的基因中，C组轮状病毒存在遗传多样性。我们的基因8探针可能类似于用于A组轮状病毒血清分型的VP7探针，在C组轮状病毒血清分型的杂交检测中有用。然而，我们对C组轮状病毒进行血清分型的遗传方法的最终确认有待对这些病毒进行血清学分析。
