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

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。

In order to prepare the polyclonal antibody of silkworm CSP9, 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-28 a 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.