蓖麻蚕NPV的增殖及其DNA分子克隆

IN-VIVO PROPAGATION AND DNA MOLECULAR CLONING OF THE NUCLEAR PALYHEDROSIS VIRUS OF PHILOSAMIA CYNTHIA RICINI

试验了接种浓度、虫龄和温度的不同组合对蓖麻蚕核型多角体病毒增殖的影响。７．３×１０＾６ＰＩＢ／ｍｌ以下的接种浓度不能使４龄上的幼虫死亡，而以７．３×１０＾７ＰＩＢ／ｍｌ量接种４龄初幼虫，病毒产量可高达１２５．８＾８ＰＩＢ／头。病毒增殖的温度峰值２５℃。以大肠杆菌质粒ｐＢＲ３２５为载体，对Ｐｃｒ ＮＰＶ ＤＮＡ ＥｃｏＲＩ酶解片段进行了分子克隆，有７个克隆带有Ｐｃｒ ＮＰＶ ＤＮＡ片段。

The studies of different combinations of virus concentrations infected the larvae of Philosamia cynthia ricini showed that the highest yield of NPV was obtained when the larvae infected in the early stage of fourth instar were fed with virus at the concentration of 7.5×107 PIBs/ml and the temparature of 25±1℃.Restriction, fragments of PcrNPV DNA digest with EcoRI restriction endon-uclease can be cloned in Escherchia coli plasmids pBR325. All recombinant DNA clones were identified by both DNA dot hybridization and southern blotting with the fragment of Pcr NPV DNA digeeted using EcoRI as a hybridization probe.