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目的 对2 株 P A G E 图型相似的 B 组轮状病毒———成人腹泻轮状病毒(adult diarrhearotavirus, A D R V) 和大白鼠乳鼠腹泻轮状病毒(rat diarrhea rotavirus, R D R V) 主要基因的相关性进行研究。方法 R T P C R 扩增 A D R V 第4 、5 、9 三个基因的c D N A 全长拷贝,并获其c D N A 克隆。用 A D R V第9 基因末端引物 R T P C R 成功扩增了 R D R Vds R N A,获得相对分子质量与 A D R V 第9 基因 P C R产物814bp 相等条带( 暂称该 R N A 模板为 R D R V 第9 片段) 及其克隆。经 Digoxigenin 标记制备 A D R V 第4 、5 、9 及 R D R V 第9 基因的c D N A 探针。用核酸点杂交、 Northern 杂交进行同源性比较。结果 获得 A D R V 第4 、5 、9 三个基因及 R D R V 第9 基因的c D N A 克隆;用 A D R V 第4 、5 、9 及 R D R V第9 基因4 个c D N A 探针进行的点杂交和 Northern 杂交结果显示,在两病毒 R N A 相应点和相应片段的位置上杂交信号都为阳性,阴性对照都为阴性。
Objective To investigate the effect of two groups of rotavirus of group B with the similar pattern of P A G E - adult diarrhearotavirus (ADRV) and rat diarrhea rotavirus (R D R V) major genes were studied. Methods RT-PCR was used to amplify full-length cD N A copies of the 4, 5, 9 genes of A D R V and cloned by c D N A. R D R Vds R N A was successfully amplified by RT-PCR with primer D 9 of A D R V to obtain an 814 bp equivalent band of the P C R product of the 9th gene of A D R V The R N A template is referred to as R D R V, fragment 9) and clones thereof. Digoxigenin labeling was used to prepare c D N A probes for A D R V Nos. 4, 5, 9 and R D R V 9 genes. Nucleotide point hybridization, Northern hybridization for homology comparison. As a result, three genes of A D R V, 4, 5 and 9, and a cDNA clone of R D R V, 9, were obtained. A D R V, 4, 5, 9 and R D R V, The dot hybridization and Northern blot analysis of c D N A probe showed that the hybridization signal was positive at the corresponding point of R N A and the corresponding fragment of both viruses, and the negative control was negative.