藏猪FRZB基因克隆及真核表达载体的构建  

Construction of Eukaryotic Expression Vector of FRZB Gene in Tibetan Pig

作　　者：王志秀 张红亮 王统苗 梁文双 张博[3] 张浩[3] WANG Zhixiu;ZHANG Hongliang;WANG Tongmiao;LIANG Wenshuang;ZHANG Bo;ZHANG Hao(College of Animal Science and Technology,Yangzhou University,Jiangsu Yangzhou 225009,China;Lhasa Agricultural and Rural Bureau,Tibet Lhasa 85000,China;College of Animal Science and Technology,China Agricultural University,Beijing 100193,China)

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009 [2]西藏自治区拉萨市农业农村局,西藏拉萨860000 [3]中国农业大学动物科学技术学院,北京100193

出　　处：《中国畜牧杂志》2020年第6期50-54,共5页Chinese Journal of Animal Science

基　　金：安徽省科技重大专项(17030701005);中央引导地方项目(YDZX20195400004426)。

摘　　要：分泌型卷曲相关蛋白3(sFRP3)由FRZB基因编码,其作为Wnt信号通路的抗拮剂,可调控动物的生长性能。本研究以藏猪肝脏组织为材料,应用PCR技术克隆得到猪FRZB基因编码序列,共编码325个氨基酸。构建FRZB基因的真核表达载体,经PCR和双酶切验证,通过脂质体转染,将重组质粒转染C2C12细胞来验证该载体的有效性。结果表明:藏猪FRZB基因的编码区长度为1 067 bp,成功构建了藏猪FRZB基因表达载体pIRES2-EGFP-FRZB,经转染C2C12细胞后产生绿色荧光,表明藏猪FRZB基因在C2C12细胞中成功表达。The secreted frizzled related protein 3(SFRP3)is encoded by the FRZB gene, and it can regulate the growth performance of pigs as an antagonist of Wnt signaling pathway. The coding sequence of pig FRZB gene was cloned by PCR from the liver tissue of Tibetan pig, encoding 325 amino acids and the eukaryotic expression vector was constructed. After PCR and double enzyme digestion verification, the recombinant plasmid was transfected into C2C12 cells by liposome transfection to test the effectiveness. The results showed that the length of coding region of FRZB gene in Tibetan pig was 1067 bp and the FRZB gene expression vector p IRES2-EGFP-FRZB was successfully constructed in Tibetan pig and transfected into C2C12 cells to produce green fluorescence signal, which indicated that the successful expression of FRZB gene in C2C12 cells.

关 键 词：藏猪 FRZB基因 C2C12细胞 真核表达载体 

分 类 号：S828.2[农业科学—畜牧学]