水牛GABA A型受体π亚基基因的克隆及其在乳腺组织中的表达

Cloning of GABRP Gene and Its Expression in Buffalo Mammary Tissue

γ-氨基丁酸(GABA)是大脑中一种重要的抑制性递质,除与兴奋的传递有关外,还在内分泌组织中发挥着重要的作用。本研究对水牛GABA A型受体的π亚基基因(GABRP)CDS序列进行了克隆,并对其进行了生物信息学分析,同时应用实时荧光定量PCR和免疫组化技术分析了GABRP基因在水牛乳腺组织中的表达情况。结果表明,应用RT-PCR技术成功获得了广西本地水牛GABRP基因序列,长为1 401bp;BLAST比对结果显示,广西本地水牛的GABRP核苷酸序列与和河流型水牛的同源性为99%,GABRP在不同物种中具有较高的保守性;GABRP蛋白主要在水牛乳腺组织的腺泡上皮细胞中表达,GABRP基因在泌乳期及非泌乳期水牛乳腺组织中都有表达,且非泌乳期GABRP基因的表达显著高于泌乳期(P<0.05)。试验结果为进一步研究GABRP基因在水牛乳腺发育过程中的功能奠定了基础。

GABA is an important inhibitory neurotransmitter in the brain,beside the function of transmissing information,which plays an important role in endocrine tissues.In this study,the CDS of buffalo GABRPgene was cloned and its biological information was analyzed,the expression of GABRP in buffalo mammary tissue was also examined by Real-time quantitative PCR and immunohistochemistry.The RT-PCR results showed that GABRP CDS fragment was obtained successfully from Guangxi local buffalo,which shared 99% with that of river buffalo,its length was 1 401 bp.The phylogenetic tree showed that GABRP had high conservation in different species.The immunohistochemistry results showed that buffalo GABRP protein located at the alveolar epithelial cells of buffalo mammary.The expression of GABRP gene in non-lactation buffalo mammary was significantly higher than that of lactation tissue（P〈0.05）.The results laid a foundation for further study the function of GABRPgene in the development of buffalo mammary gland.