分子杂交鉴定杆状病毒

Identification of Baculovirus by Dot Hybridization

利用苜蓿银纹夜蛾核型多角体病毒(Autographa californica nuclear polyhedrosis virus,AcNPV) DNA EcoRI-I片段作为同源探针,在35℃条件下,通过Southern-blot hybridization将斜纹夜蛾核型多角体病毒(Prodenia Iiturs nuclear polyhedrosis virus. PlNPV)多角体蛋白基因定位于其HindⅢ-A片段。以PlNPV DNA HindⅢ-A片段和PlNPV DNA HindⅢ-C片段分别制备探针,在不同的杂交条件下,对PINPV,甘兰夜蛾NPV(Mamestra brassicae nuclear polyhedrosis virus,MbNPV)油桐尺蠖NPV(Buzura suppresseria nuclear polyhedrosis virus,BsNPV)进行了鉴定。结果表明:由A片段制备的探针,在不同的杂交条件下,具有相应不同的非特异性,而由C片段制备的探针则具有严格的特异性。

The polyhedrin gene of Prodenia lituea nuclear polyhedrosis virus(PlNPV) was mapped on the PlNPV DNA HindⅢ-A fragment by Southern-blot hybridization with the AcNPV DNA EeoRI-Ⅰ fragment as probe under 35℃ hybridization conditions. Then, two probes were prepared from PINPV DNA HindⅢ-A and PINPV DNA HindⅢ-C fragments respectively for the identification of PINPV DNA, MbNPV DNA, BsNPV DNA under different hybridization conditions. The results showed that the probe containing the polyhedrin gene was more extensive, while the probe Jacking the polyhedrin gene more specific.