山羊PTHrP基因的多克隆抗体制备和组织表达图谱分析  

Preparation of Goat PTHrP Polyclonal Antibody and Analysis of PTHrP Expression Map in Tissues

作　　者：郑惠玲[1] 杨振宇[1] 祝珍珍[1] 安俊辉[1] 邢瑞芳[1] 

机构地区：[1]西北农林科技大学动物科技学院陕西省农业分子生物学重点实验室,杨凌712100

出　　处：《畜牧兽医学报》2011年第2期177-181,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：转基因生物新品种培育重大专项(2009ZX08009-162B);中央高校基本科研业务费专项资金(Z109021002)资助

摘　　要：旨在获得山羊PTHrP多克隆抗体,检测其在山羊各组织中的表达图谱,以便进一步研究山羊PTHrP的功能。本研究构建山羊PTHrP基因的原核表达载体,转化E.coliBL21(DE3)菌株,IPTG诱导表达获得融合蛋白,SDS-PAGE电泳检测融合蛋白,然后将融合蛋白纯化后多点免疫大耳白兔,制备多克隆抗体,最后使用West-ern blot检测PTHrP在山羊各组织中的表达。结果,酶切和测序显示原核表达载体构建成功;SDS-PAGE电泳结果说明融合蛋白(约36.5 ku)被成功纯化;间接ELISA检测所制备的抗体效价达1∶51 200;表达谱显示PTHrP在山羊乳腺、肾脏、垂体、脑、心脏、肝脏、骨、肌肉、脾脏等组织中均有表达。本研究成功制备了山羊PTHrP的多克隆抗体,发现PTHrP在山羊各组织中广泛表达。To further understand the biological function of goat PTHrP gene,the PTHrP polyclonal antibody was prepared and the tissue expression profile of PTHrP was analyzed in this study.Prokaryotic expression vector was constructed and expressed in E.coli BL21（DE3） host cells.Fusion protein was purified and used to immunize rabbit.Western blot was performed to assay the tissue expression profile.Restriction enzyme mapping and sequencing showed that prokaryotic expression vector was constructed successfully.SDS-PAGE showed that fusion protein（about 36.5 ku） was purified.The anti-PTHrP polyclonal antibody showed high sensitivity（1︰51 200） and specificity.The PTHrP protein was detected in mammary gland,kidney,pituitary,brain,heart,liver,bone,muscle and spleen.The result indicated that goat anti-PTHrP polyclonal antibody was obtained.The PTHrP expressed widely in goat tissues.

关 键 词：甲状旁腺相关肽 原核表达 多克隆抗体 组织表达图谱 山羊 

分 类 号：S827[农业科学—畜牧学] S852.4[农业科学—畜牧兽医]