经黄胫小车蝗增殖的亚洲小车蝗痘病毒DNA和结构蛋白的特性

CHARACTERIZATION OF DNA AND STRUCTURAL PROTEINSFROM OEDALEUS ASIATICUS ENTOMOPOXVIRUSPROPAGATED IN OEDALEUS INFERNALIS

从内蒙古亚洲小车蝗（Ｏｅｄａｌｅｕｓａｓｉａｔｉｃｕｓ）上分离亚洲小车蝗痘病毒（ＯｅｄａｌｅｕｓａｓｉａｔｉｃｕｓＥｎｔｏｍｏｐｏｘｖｉｒｕｓ，ＯｅａＥＰＶ），经口接种黄胫小车蝗（Ｏｅｄａｌｅｕｓｉｎｆｅｒｎａｌｉｓ）以繁殖病毒，并获得纯化的病毒包涵体。纯化的病毒包涵体经碱解、酚抽提，得到ＯｅａＥＰＶＤＮＡ。经紫外分光光度计测定，该ＤＮＡ具有典型的核酸紫外吸收光谱。根据热变性曲线测得ＯｅａＥＰＶ－ＤＮＡ的Ｔｍ值为７７．０℃，Ｇ＋Ｃ％为１７．８％，增色效应明显。该ＤＮＡ经限制性内切酶ＥｃｏＲＩ、ＢｇｌⅡ和ＨｉｎｄⅢ酶切后，分别得到１５、１６和８个片段，用片段积加法求得的分子量相似，平均为１２２．７×１０￣６道尔顿。经ＳＤＳ－ＰＡＧＥ电泳，ＯｅａＥＰＶ粒子的结构蛋白至少有４６种多肽，有５种含量较高。经８ｍｏｌ／Ｌ尿素裂解，ＳＤＳ－ＰＡＧＥ电泳，ＯｅａＥＰＶ包涵体蛋白主要由一种多肽组成，其分子量约为１０００００道尔顿。包涵体蛋白富含天冬氨酸和亮氨酸。

biochemical characterization was made on entomopoxvirus oedaleus asiaticus(OeaEPV)propagated in the grasshopper Oedaleus infernalis. The Tm of OeaEPV-DNA is about 77.0℃ based on the thermal denaturation curve. Therefore, its G+C content is about 17.8%. Athermal hyperchromic effect was apparent. The restriction endonuclease analysis revealed thatEcoRⅠ, BglⅡ and HindⅢ digested OeaEPV-DNA into 20, 23 and 11 DNA fragments,respectively. The molecular weights were comparable, with a total average value about 122.7x 10￣6 daltons. At least forty-six polypeptides were detected after separation by SDSpolyacrylamide gel electrophoresis. Five of these polypeptides were stained more intensely thanthe other polypeptide bands in the profile. The occlusion body matrix of OeaEPV, whenextracted by 8mol/L urea, had a molecular weight of approximately 100,000 daltons, and wasrich in aspartate and leucine.