家蚕化学感受蛋白BmCSP8表达谱及多克隆抗体制备  

Expression Profiling and Preparation of Polyclonal Antibody of Chemosensory Protein BmCSP8 in the Silkworm, Bombyx mori

作　　者：邓培渊[1] 李玉华[2] 袁丹丹 王国霞[2] 李长看[1] 

机构地区：[1]郑州师范学院自然博物馆,郑州450044 [2]郑州师范学院生命科学系,郑州450044 [3]河南省煤炭总医院眼科,郑州450000

出　　处：《中国农学通报》2014年第19期298-303,共6页Chinese Agricultural Science Bulletin

基　　金：国家自然基金"家蚕嗅觉受体与信息素结合蛋白相互作用的功能研究"(30700070);郑州师范学院校内课题"郑州地区主要蔬菜害虫气味结合蛋白基因功能研究"(2012082)

摘　　要：为阐明昆虫化学感受蛋白(chemosensory proteins,CSPs)基因在昆虫不同组织部位和发育时期表达特性,运用半定量RT-PCR的方法分析了家蚕BmCSP8基因的时空表达谱。结果显示:(1)BmCSP8基因在雌、雄蛹中均表达,但在卵期不表达,在幼虫阶段,低龄期高于高龄期的表达量;(2)BmCSP8基因广泛分布于成虫身体各组织,但雌雄间的表达存在差异性,在主要感受器官触角中的表达量较低;运用重组BmCSP8蛋白成功制备兔多克隆抗体,Western-blot分析发现抗血清与触角蛋白发生特异性结合,为深入研究其生理功能打下基础。以上结果表明,BmCSP8基因可能参与了家蚕觅食的生理过程,且具有其他尚未明确的非嗅觉功能。In order to clarify the expression characteristics of chemosensory protein（CSP） gene in differenttissue parts and developmental stages in insect, the spatio- temporal expression profile of BmCSP8 wasinvestigated using semi-quantitative RT-PCR. The results showed that：（1） BmCSP8 was expressed in larvaand pupa stage, but not in egg stage. In larva stage, the expression was more in younger stages than that inolder stages.（2） BmCSP8 was expressed in various adult tissues including antenna, head（without antenna）,wing, leg, thorax and abdomen. But there were differences existing between female and male, and theexpression level was low in antenna which was the primary sense organs of insect.（3） Rabbit polyclonalantibody was prepared successfully by recombinant BmCSP8 protein, Western- blot anlysis showed thatantiserum could combined specifically with the antenna protein, it also provided a foundation for furtherresearch on the BmCSP8 physiological functions. These results suggested that BmCSP8 may has unknown non-olfactoryfunctions, and most likely involved in thephysiological process of foraging.

关 键 词：家蚕 化学感受蛋白 半定量RT-PCR 蛋白纯化 多克隆抗体 

分 类 号：Q966[生物学—昆虫学] Q786