Expression of 2．6kb Enhancin Gene Fragment from Helicoverpa atmigera Granulosis Virus

HU Rong; MENG Xiao-Lin; XU Jin; WANG Jian; LIJ Wei

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August 2001

HU Rong, MENG Xiao-Lin, XU Jin, WANG Jian and LIJ Wei. Expression of 2．6kb Enhancin Gene Fragment from Helicoverpa atmigera Granulosis Virus[J]. Virologica Sinica, 2001, 16(4): 364-368.

Citation: HU Rong, MENG Xiao-Lin, XU Jin, WANG Jian, LIJ Wei. Expression of 2．6kb Enhancin Gene Fragment from Helicoverpa atmigera Granulosis Virus .VIROLOGICA SINICA, 2001, 16(4) : 364-368.

棉铃虫颗粒体病毒增效蛋白基因2．6kb片段的表达

1.
武汉大学病毒研究所，湖北武汉430072

摘要

以棉铃虫颗粒体病毒 (Helicoverpaarmigeragranulosisvirus ,简称HaGV)基因组的DNA为模板设计引物 ,PCR扩增病毒增效蛋白 (Enhanicn)基因 ,然后经BamHI/PstI双酶切消化 ,得到近乎全长的约 2 .6kb的增效蛋白基因片段 ,再与pQE32质粒连接 ,构建了重组表达载体pQE32 /En ,转化大肠杆菌M15 (pREP4 ) ,在IPTG诱导下表达出分子量约为 10 2kD的融合蛋白并命名为P10 2 ,纯化的P10 2 包涵体显示了明显的增效活性 ,在感染后 16 8h时统计可提高HaNPV对棉铃虫幼虫的感染死亡率 6 .2 5 %～ 2 7.0 9% ,缩短LT50 12 .3h以上 ;在感染后 72h时统计可提高Bt对棉铃虫幼虫的感染死亡率 2 8.18% ,缩短LT50 12 .33h。The 2.6kb fragment of enhancing gene from Helicoverpa armigera granulosis virus was inserted into vector pQE32 and expressed successfully in E.coli M15.The synergy of expression product(P 102 ) on Helicoverpa armigera nuclear polyhedrosis virus(HaNPV)and Bt against the 2nd instar larvae of H.armigera was studied.The results indicated that the accumulated mortality of the larvae increased 6.25%～27.09% on 168h post infection and the median lethal time decrease at least 12.3h in HaNPV+En treated compared with those in HaNPV treated.The accumulated mortality of the larvae increase 28.18% on 72h postinfection and the median lethal time decreased 12.33h in Bt+En treated compared with that in Bt treated.
- 棉铃虫颗粒体病毒
- , 增效蛋白基因
- , 克隆
- , 表达
- , 生物活性测定
- , Helicoverpa
- , armigera
- , Granulosis
- , Virus
- , Enhancin
- , Cloning
- , Expression
- , Bioassay

Expression of 2．6kb Enhancin Gene Fragment from Helicoverpa atmigera Granulosis Virus

1.
The 2.6kb fragment of enhancing gene from Helicoverpa armigera granulosis virus was inserted into vector pQE32 and expressed successfully in E.coli M15.The synergy of expression productP 102 on Helicoverpa armigera nuclear polyhedrosis virusHaNPVand Bt against the 2nd instar larvae of H.armigera was studied.The results indicated that the accumulated mortality of the larvae increased 6.25%～27.09% on 168h post infection and the median lethal time decrease at least 12.3h in HaNPV+En treated compared with those in HaNPV treated.The accumulated mortality of the larvae increase 28.18% on 72h postinfection and the median lethal time decreased 12.33h in Bt+En treated compared with that in Bt treated.

Abstract

The 2.6kb fragment of enhancing gene from Helicoverpa armigera granulosis virus was inserted into vector pQE32 and expressed successfully in E.coli M15.The synergy of expression product(P 102 ) on Helicoverpa armigera nuclear polyhedrosis virus(HaNPV)and Bt against the 2nd instar larvae of H.armigera was studied.The results indicated that the accumulated mortality of the larvae increased 6.25%～27.09% on 168h post infection and the median lethal time decrease at least 12.3h in HaNPV+En treated compared with those in HaNPV treated.The accumulated mortality of the larvae increase 28.18% on 72h postinfection and the median lethal time decreased 12.33h in Bt+En treated compared with that in Bt treated.
- Helicoverpa armigera Granulosis Virus Enhancin Cloning Expression Bioassay

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression of 2．6kb Enhancin Gene Fragment from Helicoverpa atmigera Granulosis Virus

1. The 2.6kb fragment of enhancing gene from Helicoverpa armigera granulosis virus was inserted into vector pQE32 and expressed successfully in E.coli M15.The synergy of expression productP 102 on Helicoverpa armigera nuclear polyhedrosis virusHaNPVand Bt against the 2nd instar larvae of H.armigera was studied.The results indicated that the accumulated mortality of the larvae increased 6.25%～27.09% on 168h post infection and the median lethal time decrease at least 12.3h in HaNPV+En treated compared with those in HaNPV treated.The accumulated mortality of the larvae increase 28.18% on 72h postinfection and the median lethal time decreased 12.33h in Bt+En treated compared with that in Bt treated.

Abstract: The 2.6kb fragment of enhancing gene from Helicoverpa armigera granulosis virus was inserted into vector pQE32 and expressed successfully in E.coli M15.The synergy of expression product(P 102 ) on Helicoverpa armigera nuclear polyhedrosis virus(HaNPV)and Bt against the 2nd instar larvae of H.armigera was studied.The results indicated that the accumulated mortality of the larvae increased 6.25%～27.09% on 168h post infection and the median lethal time decrease at least 12.3h in HaNPV+En treated compared with those in HaNPV treated.The accumulated mortality of the larvae increase 28.18% on 72h postinfection and the median lethal time decreased 12.33h in Bt+En treated compared with that in Bt treated.