棉铃虫颗粒体病毒增效蛋白基因5′端截短片段的表达及增效活性测定  

EXPRESSION OF TWO TRUNCATED ENHANCIN GENE FROM HELICOVERPA ARMIGERA GRANULOSIS VIRUS IN E.COLI AND ITS RELIMINARY BIOASSAY *

作　　者：刘相国[1] 杨恭[1] 邱并生[1] 张克勤[2] 田波[1] 

机构地区：[1]中国科学院微生物研究所分子病毒学和生物工程开放实验室,北京100080 [2]云南大学工业微生物重点实验室,昆明650091

出　　处：《微生物学报》2001年第2期167-172,共6页Acta Microbiologica Sinica

基　　金：国家自然科学基金!( 939970 50 3);云南省计委高技术项目!( 1 699 2 62 )资助&&

摘　　要：将带有棉铃虫颗粒体病毒增效蛋白 (HaGVenhancin)基因的重组表达载体 pET 30a En分别用BalⅠ和DraⅠ酶切后连接 ,构建重组表达载体pET 30a Ben和 pET 30a Den ,其外源基因的开放阅读框分别为HaGV增效蛋白基因 5′端的 1 7kb和 2 2kb。IPTG诱导表达产生 66 7kD和 85 1kD的多肽并命名为Ben和Den。初步纯化的Ben和Den显示了对棉铃虫核多角体病毒 (HaNPV)及Bt的增效活性。Ben可对棉铃虫核多角体病毒增效 1 0 5%～2 6 5% ,LT50 缩短 0 9d ,Den增效 1 0 2 %～ 33 0 % ,LT50 缩短 1 2d～ 1 9d。重组增效蛋白可对Bt(1∶2 50 0稀释 )增效 2 0 7%～ 35 4% ,Den增效 1 6 7%～ 31 5% ,Ben增效 1 1 7%～2 7 4%。The plasmids pET 30a\|Ben and pET\|30a\|Den which included 1 7kd and 2 2kb fragments of 5′\|terminal of HaGV enhancin gene were obtained by cutting recombinant plasmid pET\|30a\|En with Bal Ⅰ and Dra Ⅰ respectively.Two fragments were expressed in E.coli successfully and the products were named Ben and Den respectively.The enhancement,which Ben and Den enhance the infectivity of HaNPV and Bt in 3 rd larvae of Helicoverpa armigera, was studied.The results indicated that there was increase of the mortality of 10 5%～26 5% and the LT 50 decrease of 0 9d causes by adding Ben,while Den could increase the mortality by 10 2%～33 0% and decrease the LT 50 by 1 2d～1 9d.The preliminary bioassay on Bt against Helicoverpa armigera indicated the recombinant enhancin could increase the mortality of larvae by 207%～354%\,Den by 16 7%～315%\,Ben by 11 7%～27 4%.

关 键 词：棉铃虫 颗粒体病毒 增效蛋白 HANPV BT 增效活性 昆虫病毒 

分 类 号：S432.4[农业科学—植物病理学]