摘要
旨在克隆猪lipasin基因,并检测其对肝细胞LPL基因转录的影响。本试验利用同源电子克隆技术克隆猪lipasin基因,构建其真核表达载体转入肝细胞,探讨其对LPL基因转录的影响。结果表明,成功克隆了猪lipasin基因,目的片段大小为615bp,上传至GenBank(登录号:KF017598),开放阅读框为594bp,编码197个氨基酸,liapsin蛋白的分子量和等电点分别为21.99ku和6.79;成功构建了真核表达载体PB-EGFP-lipasin,实现lipasin基因在肝细胞中的过表达,与对照组相比,PB-EGFP-lipasin转染的细胞中LPL基因的转录水平显著下降。结果提示,猪lipasin可能通过抑制肝细胞LPL的转录影响脂代谢。
The study aimed to clone the lipasingene and investigate its effects on LPLgene transcription in liver cell in pig.The porcine lipasingene was obtained by using homologous electronic cloning techniques and the eukaryotic expression vector was constructed and transfected into hepatocytes,and further to identify its effect on the transcription level of LPL gene.The results showed that lipasingene was successfully cloned,the segment size was 615 bp,it was deposited to GenBank with the accession number KF017598.The size of open reading frame（ORF）of lipasin was 594 bp and it encoded 197 amino acids.The weight of lipasin molecular and theoretical isoelectric point were 21.99 ku and 6.79,respectively.The recombinant eukaryotic expression vector PBEGFP-lipasin was successfully constructed and transfected into hepatocyte,realizing the overexpression of lipasingene.Compared with the control group,the transcription level of LPL mRNA decreased significantly in PB-EGFP-lipasin transfected hepatocyte.These results suggest that lipasin may affect lipid metabolism by inhibiting the LPLgene transcription in hepatocyte.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2016年第2期411-416,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
农业部"948"重点计划(2011-G35)
农业部转基因重大专项(2014ZX0801015B)
河南省重点科技攻关项目(112102310705)
