家蚕化学感受蛋白CSP9的抗体制备及表达分析

doi:10.11924/j.issn.1000-6850.casb20190400023

中国农学通报 ›› 2019, Vol. 35 ›› Issue (34): 44-49.doi: 10.11924/j.issn.1000-6850.casb20190400023

所属专题：生物技术

家蚕化学感受蛋白CSP9的抗体制备及表达分析

魏博帆,王景娅,孙妍妍,王露,李晓哲,邱礽,乔惠丽

南阳师范学院农业工程学院河南省伏牛山昆虫生物学重点实验室

收稿日期:2019-04-22 修回日期:2019-11-19 接受日期:2019-07-23 出版日期:2019-12-09 发布日期:2019-12-09
通讯作者: 乔惠丽
基金资助:
河南省高等学校青年骨干教师项目“家蚕化学感受蛋白CSP16 在蜕皮过程中的功能研究”(NO.2016GGJS-127)；河南省科技厅科技攻关项目“用于非编码RNA功能研究的CRISPR-Cas9 技术体系构建”(NO.182102110447)；河南省高等学校重点科研项目“家蚕巨噬细胞移动抑制因子（MIF）在BmNPV感染中的作用研究”(NO.18A180024)。

Silkworm Chemosensory Protein CSP9: Antibody Preparation and Expression Analysis

王景娅, 孙妍妍

Received:2019-04-22 Revised:2019-11-19 Accepted:2019-07-23 Online:2019-12-09 Published:2019-12-09

摘要/Abstract

摘要： 为了制备家蚕CSP9的多克隆抗体，研究该蛋白在家蚕中的表达特征和功能。利用PCR扩增家蚕csp9基因，构建其原核表达载体，诱导CSP9蛋白表达并纯化。纯化后的CSP9蛋白免疫新西兰大白兔制备多克隆抗体，Western杂交检测CSP9在家蚕中的表达特征。最终成功构建csp9/pET-28a原核表达载体，并纯化获得与预测分子量一致的CSP9蛋白；成功制备了CSP9多克隆抗体；Western杂交结果表明，CSP9在家蚕不同时期和不同组织中都有特异表达。本研究成功表达纯化并制备了家蚕CSP9多克隆抗体，分析了CSP9在家蚕中的表达特征，为后续该蛋白在家蚕中的功能和调控研究奠定了基础。

关键词: 藜麦, 藜麦, 种质资源, 适应性种植

Abstract: In order to prepare the polyclonal antibody against CSP9 of silkworm, and study the expression characteristics and functions of CSP9 in silkworm. The csp9 gene of silkworm was amplified by PCR, and its prokaryotic expression vector was constructed. The expression of CSP9 protein was induced and purified. The purified CSP9 protein was immunized to New Zealand purebred rabbit to prepare polyclonal antibody. The expression of CSP9 in silkworm was detected by Western blot. Finally, the prokaryotic expression vector of csp9/pET-28a was constructed successfully, and the CSP9 protein with predicted molecular weight was purified. The polyclonal antibody against CSP9 was successfully prepared. The result of Western blot showed that CSP9 was specifically expressed in different stages and tissues of silkworm. In this study, CSP9 protein was successfully expressed and purified, its polyclonal antibody was successfully prepared. The expression characteristics of CSP9 in silkworm were analyzed, which laid a foundation for further study on the function and regulation of CSP9 in silkworm.

中图分类号:

Q812

魏博帆,王景娅,孙妍妍,王露,李晓哲,邱礽,乔惠丽. 家蚕化学感受蛋白CSP9的抗体制备及表达分析[J]. 中国农学通报, 2019, 35(34): 44-49.

王景娅,孙妍妍,,, and . Silkworm Chemosensory Protein CSP9: Antibody Preparation and Expression Analysis[J]. Chinese Agricultural Science Bulletin, 2019, 35(34): 44-49.

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家蚕化学感受蛋白CSP9的抗体制备及表达分析

Silkworm Chemosensory Protein CSP9: Antibody Preparation and Expression Analysis

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