猪H-FABP基因的克隆及真核表达载体的构建  

Cloning and Constructing of Eukaryotic Expression Vector of H-FABP Gene in Pig

作　　者：戚顺民 马康[1] 吴明明[2] 孙金海[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]中国农业大学,北京100193

出　　处：《中国畜牧杂志》2015年第11期69-73,共5页Chinese Journal of Animal Science

基　　金：转基因生物新品种培育科技重大专项(2013ZX08006-003);山东省现代农业产业技术体系生猪创新团队建设项目(SDAIT-06-022-01)

摘　　要：心肌脂肪酸结合蛋白基因(H-FABP)是影响家猪肌内脂肪含量的重要候选基因之一。本研究以长白猪为实验材料,采用RT-PCR方法克隆H-FABP基因全长编码序列,并构建pEGFP-N1-H-FABP融合表达载体,通过脂质体介导转染PK15细胞,48 h后进行荧光表达检测。获得了猪H-FABP基因去终止密码子全长编码序列399 bp,成功构建pEGFP-N1-H-FABP融合表达载体,瞬时转染后在PK15细胞中顺利表达。本实验为进一步从细胞水平上解析及验证该基因的生物学功能奠定了基础。Heart fatty acid binding protein gene （H-FABP） is one of the most important candidate genes that affect pig＇s intramuscular fat content. In this research, the full-length coding sequence of H-FABP gene was ch）ned from Landrace by RT-PCR method, pEGFP-N1-H-FABP fusion expression vector was eonstrtn＇ted and detected by liposome-medialed transfection PKI5 cells with fluorescence expression after 48 h. The whole CDS size of H-FABP was 399 hp excluding the termination codon. The pEGFP-NI-H-FABP fusion expression vector was constructed and successfully expressed in PK15 cells. The results of this trial provide a basis for parsing and verifying the biological function of this gene on cellular level.

关 键 词：H-FABP 真核表达 融合表达 长白猪 

分 类 号：S828.2[农业科学—畜牧学]