南方菜豆花叶病毒(SBMV)两典型株系特异cDNA和RNA探针的制备及应用

THE APPLICATION AND PREPARATION OF THE STRAIN SPECIFIC cDNA AND RNA PROBES OF SOUTHERN BEAN MOSAIC VIRUS

南方菜豆花叶病毒（ＳＢＭＶ）主要有２株系：菜豆株系（ＳＢＭＶ－Ｂ）和豇豆株系（ＳＢＭＶ－Ｃ），血清学方法不能予以区分。根据已报道的核酸序列设计了２对特异引物，进行ＲＴ－ＰＣＲ扩增并克隆到载体Ｂｌｕｅｓｃｒｉｐｔ中，序列测定予以证实后利用随机引物法合成放射性ｃＤＮＡ探针，再进行体外转译合成地高辛ＵＴＰ标记的ＲＮＡ探针。２种探针均可分别特异地检测Ｎｏｒｔｈｅｒｎｂｌｏｔ膜上的ＳＢＭＶ－Ｂ和ＳＢＭＶ－Ｃ。ＲＮＡ探针检测ＳＢＭＶ－Ｂ和ＳＢＭＶ－Ｃ灵敏度分别为：０．１μｇ和０．０１μｇ。

As serological techniques could not distinguish SBMV B and SBMV C,two type strains of southern bean mosaic virus,the fragments of the nucleic acid of the two strains were cloned and the specific radioactive cDNA and digoxigenin UTP labeled RNA probes were prepared.Both of the probes could specifically detect SBMV B SBMV C respectively on northern blot,as little as 0.1μg of SBMV B and 0.01μg of SBMV C were visualized.