鸡黑素皮质素受体-4多肽抗体的制备与鉴定  

Preparation and identification of melanocortin-4 receptor peptide antibody

作　　者：郎斌[1] 朱弘焱[1] 王佳美[1] 苏玉虹[1] 田玉民[1] 

机构地区：[1]辽宁医学院畜牧兽医学院,辽宁锦州121000

出　　处：《畜牧与兽医》2013年第9期16-21,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31172242)

摘　　要：根据GenBank中报道的鸡黑素皮质素受体-4(melanocortin-4 receptor,MC4R)一级结构信息,用TMHMM 2.0和DNAstar软件分析MC4R蛋白的跨膜结构域、二级结构、疏水性、亲水性以及抗原性等,综合考虑抗体设计的其他因素,设计出1段15个氨基酸的多肽。将合成后的多肽与牛血清白蛋白(BSA)偶联,用与BSA偶联的MC4R多肽免疫新西兰兔,3个月后获取血清,亲和纯化出抗MC4R多肽抗体。通过ELISA法检测效价,Western blot和免疫组化法检测抗体特异性。通过该方法得到了高效价与高特异性的鸡MC4R多肽抗体,ELISA法测定其效价可达到1∶128 000,而且该抗体可用于免疫组化,说明所制备的鸡MC4R抗体具有高特异、高效价等特点,将为鸡MC4R基因的功能研究提供物质基础。According to the primary structure information in GenBank and the prediction of the possible conformational structure, hydropho- bicity and antigenicity of chicken melanocortin-4 receptor （MC4R） and the principal for antibody production with TMHMM 2. 0 and DNAs- tar software, a partial peptide with 15-amino acid residues of MCAR was synthesized. The synthesized peptide was then used to immunize New Zealand rabbit after coupled with albumin from bovine serum （BSA）. After the MCAR peptide antibody was purified, the titer and specificity of anti-MCAR and peptide antibody were determined by ELISA, Western blot and immunohistochemical analysis. The results showed the chicken MC4R peptide antibody with a high titre of 1 ： 128000 was obtained. And the antibody could be used in the immunohistochemica1 examination. This study provides useful material for investigation of function on MC4R gene.

关 键 词：鸡 MC4R 多肽抗体 特异性 

分 类 号：S852.2[农业科学—基础兽医学]