检测烟草中烟草花叶病毒的RNA斑点杂交法

用普通烟草花叶病毒OM株3′-端约2kb的cDNA为探针,探索了用RNA斑点杂交法对烟草组织中烟草花叶病毒RNA进行检测的条件。这些条件包括用分子杂交法观察云南烟区和上海烟草上分到的烟草花叶病毒与OM株的同源性,从烟草组织中提取烟草花叶病毒的几种方法的比较,使RNA有效地固定在硝酸纤维素滤膜上的方法,烟草组织中是否有干扰RNA固定和杂交的物质,斑点杂交方法检测烟草花叶病毒的特异性、灵敏度等。

RNA DOT HYBRIDIZATION FOR DETECTING TOBACCO MOSAIC VIRUS IN TOBACCO LEAVE

The conditions for detecting tobacco mosaic virus (TMV) RNA in tobacco leave extract by RNA dot hybridization using a radioactive probe containing the cDNA of 3' terminal RNA of TMV strain OM have been studied. These conditions include the homology between the TMV strains isolated from Yunnan and Shanghai and the strain OM, the method for extracting viral RNA from tobacco leave and for adsorbing the extracted viral RNA onto nitrocellulose filter efficiently, the material interrupting the adsorption of viral RNA onto nitrocellulose filter and the hybridization of viral RNA with probe, and the speciality, sensitivity of the RNA dot hybridization method for detecting TMV-RNA in tobacco leave.