促性腺激素抑制激素—抑制素—卵泡抑制素融合基因疫苗的构建  

Construction of the Gonadotropin Inhibiting Hormone-Inhibiting Hormone-Follicostatin Fusion Gene Vaccine

作　　者：王天翔 李范文 刘继刚 王喜军 王丽娟 吕会芹 王洮生 

机构地区：[1]甘肃省绵羊繁育技术推广站,甘肃张掖734000

出　　处：《畜牧兽医杂志》2016年第5期9-12,16,共5页Journal of Animal Science and Veterinary Medicine

基　　金：国家绒毛用羊产业技术体系(CARS-40-33)

摘　　要：促性腺激素抑制激素(GnIH)是唯一被阐明对GnRH、LH起着负调控作用的神经肽,在非繁殖期抑制动物的繁殖功能,为运用生殖免疫原理中和GnIH,调节GnRH、LH激素水平,诱导藏羊非繁殖季节发情提供契机。本研究以GnIN-INH-Follistatin-(C3d)融合的PET28a-GnIN-INHFollistatin-(C3d3)原核表达载体为基础,将GnIN-INH-Follistatin-C(3d3)转入真核疫苗表达载体pSEC-tag2a中构建pSEC-tag2a-GnIN-INH-Follistatin-C(3d3)基因疫苗。将质粒转染HEK293细胞,转染24h后收集细胞,做采用takara的荧光定量PCR用2Xpremix对基因进行RT-PCR试验来验证,表明pSEC-tag2a-GnIN-INH-Follistatin-C3d3质粒转染HEK293细胞后能够在HEK293细胞内稳定表达。结果表明,成功构建INH-GnIH融合基因疫苗,为开展研究免疫后非繁殖季节藏羊生殖激素表达规律工作提供理论和技术支撑。Gonadotropin inhibiting hormone is the only elucidated neuropeptide that plays a negative regulatory role on gonadotropin-releasing hormone and luteotropic hormone.As a result,it can inhibit animal reproduction function during nonbreeding period.This study provide an opportunity for inducing the Tibetan estrus at anestrous season by reproductive immune principle to neutralization gonadotropin inhibiting hormone,adjusting the gonadotropin-releasing hormone and luteotropic hormone.With the GnIH-INH-Follistatin fusion of pET28a-GnIH-INH-Follistatin-3（C3d）prokaryotic expression vector as the foundation,then GnIH-INH-Follistatin-3（Cd3）was transferred into eukaryotic expression vector vaccine pSEC-tag2 ato build the vaccineof pSEC-tag2a-GnIH-INH-Follistatin-3（Cd3）gene.With the transfectionof plasmid to HEK293 cells,after 24 hours,collecting cells,then doing the takara fluorescence quantitative PCR in 2Xpremix for gene RT-PCR test to verify.The results showed that pSEC-tag2a-GnIH-INH-Follistatin-C3d3 plasmid can be stably expressed in HEK293 cells after transfection.The results revealed that the INH-GnIH fusion gene vaccine was established,which lay a foundation for the multipleting immunization work in the non-breeding season.

关 键 词：促性腺激素抑制激素 抑制素 基因疫苗 

分 类 号：S859.796[农业科学—临床兽医学]