摘要
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。
Cloning of the major gene segments of group B rotavirus ADRV and RDRV and their comparison$$$$ YANG Hongyan*, JI Shaozhong, KANG Zhigang, et al. *Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing 102206 【Abstract】 Objective To compare the major gene segments of group B rotavirus adult diarrhea rotavirus (ADRV) and infant rat diarrhea rotavirus (RDRV) which share the same RNAs migration pattern on polyacrylamide gel electrophoresis (PAGE). Methods The full length cDNA of ADRV gene 4, 5, 9 were produced by reverse transcription and polymerase chain reaction (RTPCR) from dsRNA of ADRV with terminal primers of each gene segments, respectively. And the above cDNAs were cloned. The dsRNA of RDRV was amplified successfully with the terminal primer of ADRV gene 9. The molecular weight of the RDRV PCR product was same as that of ADRV gene 9s 814bp (the RNA templet was called RDRV segment 9 temporarily) and the clone was demonstrated. The four ADRV gene 4,5,9 and RDRV gene 9 full length cDNA probes were made from aimed digested gene segments by labelling with Digoxigenin. Genic identity was studied with Dot and Northern hybridization. Results The full length cDNA clones of ADRV gene 4,5,9 and RDRV gene 9 were obstained. The hybridization signals of ADRV and RDRV RNA homologizing dots and fragments were all positive and the every negative control was negative. Conclusion RDRV share high identity with ADRV identified with Dot and Northern blot hybridization using the probes of ADRV gene 4,5,9 and RDRV gene 9. 【Subject words】 Adult diarrhea rotavirus Rat diarrhea rotavirus Genic identity Gene,viral
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1999年第5期364-368,共5页
Chinese Journal of Microbiology and Immunology
关键词
腹泻轮状病毒
基因同源性
病毒基因
Adult diarrhea rotavirus
Rat diarrhea rotavirus
Genic identity
Gene
viral
