A bacterially produced virus enhancing factor from an entomopoxvirus enhances nucleopolyhedrovirus infection in armyworm larvae.
A bacterially produced virus enhancing factor from an entomopoxvirus enhances nucleopolyhedrovirus infection in armyworm larvae.
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DOI：
10.1006/jipa.2001.5036
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发表时间：
2001-07
影响因子：
3.4
通讯作者：
T. Hukuhara;T. Hayakawa;A. Wijonarko
中科院分区：
文献类型：
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作者：
T. Hukuhara;T. Hayakawa;A. Wijonarko
利用大肠杆菌表达系统pGEX-2 T，表达了一种能增强粘虫多核型多角体病毒（PsunMNPV）感染粘虫幼虫的病毒增强因子（EF）。转化E.大肠杆菌细胞，这是不活跃的增强PsunMNPV感染，变得活跃时，用胰蛋白酶或凝血酶处理。裂解物中的GST-EF融合蛋白用本体GST纯化模块纯化，并用凝血酶切割成EF和GST部分。用谷胱甘肽-琼脂糖凝胶4 B去除GST部分导致高度纯化的EF制剂，其增强了PsunMNPV在粘虫幼虫中的感染以及PsunMNPV与粘虫细胞系SIE-MSH-805-F的融合。
Using an Escherichia coli expression system, pGEX-2T, that expresses foreign sequences as fusion proteins with a glutathione S-transferase (GST) carrier, we have expressed a virus enhancing factor (EF) from Pseudaletia separata entomopoxvirus, which enhances P. unipuncta multi nucleopolyhedrovirus (PsunMNPV) infection in larvae of the armyworm, P. separata. The lysates of transformed E. coli cells, which were not active in enhancing PsunMNPV infection, became active when treated with either trypsin or thrombin. The GST-EF fusion protein in a lysate was purified with a bulk GST purification module and cleaved into the EF and GST moieties with thrombin. Removal of the GST moiety with glutathione-Sepharose 4B resulted in a highly purified EF preparation, which enhanced PsunMNPV infection in armyworm larvae and PsunMNPV fusion with an armyworm cell line, SIE-MSH-805-F.