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家蚕浓核病毒BmDNV-1(伊那株)结构蛋白基因VP4的克隆、表达及抗体制备

Cloning,expression and antibody preparation of Bombyx mori densovirus(BmDNV-1)structural gene VP4
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摘要 利用PCR技术扩增出BmDNV-1结构蛋白vp4基因,并将该基因与原核表达载体pMALc2X进行连接,转化大肠杆菌DH10B,获得重组质粒经IPTG诱导表达得到大小为96 000的融合蛋白,融合蛋白经Amylose柱纯化,使用其免疫新西兰大白兔,制备出多克隆抗体.从而为进一步研究该病毒结构蛋白基因的转录和翻译机制提供可靠的工具. Bombyx mori densovirus （BmDNV-1）, based on the previously reported genome sequence, constitutes by itself a separate genus （Iteravirus） within the Densovirinae subfamily of the family parvoviridae. It is a perfect model virus to study gene regulation because of its simple genomic structure which consisted of only structure protein and non-structure protein genes. We reported here that the structural protein gene VP4 of the BrnDNV-1 was amplified by PCR and inserted into prokaryotic expression vector pMAL-c2X and then expressed in Escherichia coli DH10B strain. The target fusion protein with molecular weight of 96 000 was induced by IPTG and purified by Amylose affinity chromatography. The purified VP4 protein was used to immunize the New Zealand rabbit to produce an antibody against this protein.
作者 谢马莉 于倩 李京京 李毅
机构地区 华中师范大学生命科学学院昆虫学研究所
出处 《华中师范大学学报（自然科学版）》 CAS CSCD 北大核心 2009年第1期103-107,共5页 Journal of Central China Normal University：Natural Sciences
基金 国家自然科学基金项目(30670081)
关键词 家蚕浓核病毒 结构蛋白 克隆 表达 蛋白质纯化 多克隆抗体 Bombyx mori densovirus structural protein gene cloning expression protein purification antibody preparation
分类号 Q786 [生物学—分子生物学]
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参考文献17

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二级参考文献22

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华中师范大学学报（自然科学版）
2009年 第1期

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