家鸡生长激素释放激素受体(GHRHR)的蛋白表达与抗体制备

本研究采用RT-PCR等方法,构建编码家鸡两个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,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.

Production of recombinant chicken GHRHR1 and GHRHR2 proteins and preparation of the antibodies against chicken GHRHRs

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.