家鸡生长激素释放激素受体(GHRHR)的蛋白表达与抗体制备
Production of recombinant chicken GHRHR1 and GHRHR2 proteins and preparation of the antibodies against chicken GHRHRs

家鸡生长激素释放激素受体（GHRHR）属于G蛋白偶联受体B-I家族,其主要作用是介导生长激素释放激素(GHRH)对垂体生长激素合成与分泌的刺激效应。本研究采用RT-PCR等方法，构建了编码家鸡两个GHRH受体(GHRHR1和GHRHR2) N-末端胞外结构域的原核表达载体，并在细菌中成功诱导其表达。利用离子亲和层析技术，纯化了两受体的重组蛋白片段。 在此基础上，利用重组蛋白在兔中制备了两受体的抗体，并采用Western blot 方法，初步验证这些抗体对垂体以及HEK293细胞中表达的家鸡GHRHR1和GHRHR2识别的有效性。 这些结果为后续探究GHRHRs在家鸡组织中的细胞定位和生理功能奠定基础。
Growth hormone-releasing hormone (GHRH) receptor (GHRHR) belongs to the G protein-coupled receptor (GPCR) B-I subfamily, which is capable of mediating the actions of GHRH on pituitary growth hormone synthesis and release. In the study, we constructed the prokaryotic expression plasmids encoding the N-terminal fragment of chicken GHRHR1 and GHRHR2 and induced their expression in E. coli by IPTG treatment. Moreover, the recombinant GHRHR1 and GHRHR2 fusion proteins were purified and used to prepare the antibodies against chicken GHRHR1 and GHRHR2 in rabbits. In addition, the specificity of the antibodies against chicken GHRHRs was evaluated by Western blot analysis. The results from this study pave the way for further investigation of the localization and roles of GHRHRs in chicken tissues