关键词: 烟草花叶病毒辽宁分离物, RNA探针, DNA探针, 点印迹杂交, Northern杂交

Abstract: In order to establish a more sensitive and specific molecular hybridization detection system of Tobacco mosaic virus Liaoning isolate（TMV-LN）,total viral RNA was extracted from purified TMV-LN viral particles and amplified by reverse-transcript PCR. The PCR products were ligated into pUC119 vector to construct pUCTMV-PP that express RNA detection probe using Digoxigenin（DIG）Northern Starter Kit. DNA probe was also constructed using DIG DNA hybridization kit. Dot-blot hybridization and Northern blot hybridization analysis were performed to study the specificity and sensitivity of the RNA probe and DNA probe constructed above. Both of the probes showed high specificity and sensitivity in TMV-LN detection through Dot-blot hybridization and Northern blot hybridization detection system. Comparison of two hybridization systems,Dot-blot hybridization system is suitable for virus qualitative detection,whereas,Northern blot hybridization has its advantage in relative quantification of viral genome RNA.

Key words: TMV Liaoning isolate, RNA probe, DNA probe, Dot-blot hybridization, Northern blot hybridization