中国畜牧兽医2017，Vol.44Issue(7)：1954-1960,7.DOI:10.16431/j.cnki.1671-7236.2017.07.008
从江香猪ApoA1基因的克隆及亚细胞定位研究
Cloning and Subcellular Localization of ApoA1 Gene in Congjiang Xiang Pig
摘要
Abstract
This experiment was aimed to clone apolipoprotein A1(ApoA1) gene of Congjiang Xiang pig, and study the subcelluar localiztion of ApoA1 gene in eukaryocyte.The recombination plasmid pEGFP-C1-ApoA1 was constructed with RT-PCR and other methods, and detected by colony PCR,double digestion and sequencing, after successful construction of the recombination plasmid pEGFP-C1-ApoA1,the subcellular localization of ApoA1 protein were analyzed by fluorescence co-localization technique in the 36 h-transfected HEK-293T cells.Compared with ApoA1 gene of Sus scrofa submission in GenBank, the results showed that six base mutations were found in ApoA1 gene of Congjiang Xiang pig, five of above mentioned mutations were sense mutations, causing alanine to glutamic acid, histidine to glutamine, valine to leucine and aspartic acid to glycine in 180,185,186 and 209 amino acid residues, respectively.Using PSOR Ⅱ Prediction and fluorescence co-localization, it was found that the expression of ApoA1 protein was observed mainly in the extracellular matrix (77.8%).In conclusion, ApoA1 gene of Congjiang Xiang pig was cloned successfully, and the expression of ApoA1 protein was mainly concentrated in the extracellular matrix.These results would provide a knowledge for further constructing the ApoA1 gene transgenic animal models, and contribute to understanding the relation between ApoA1 gene and the human obesity-induced diseases.关键词
丛江香猪/ApoA1基因/pEGFP-C1载体/HEK-293T细胞/生物信息学分析/亚细胞定位Key words
Congjiang Xiang pig/ApoA1 gene/pEGFP-C1 vector/HEK-293T cell/bioinformatics analysis/subcellular localization分类
生物科学引用本文复制引用
赵佳福,段志强,杨远青,嵇辛勤,王圆圆,孙成娟..从江香猪ApoA1基因的克隆及亚细胞定位研究[J].中国畜牧兽医,2017,44(7):1954-1960,7.基金项目
省校联合基金(黔科合LH字[2014]7668) （黔科合LH字[2014]7668）
贵州省科技重大专项(黔科合重大专项字[2013]6008号) （黔科合重大专项字[2013]6008号）
贵州大学青年基金(贵大自青基合字[2012]010号) （贵大自青基合字[2012]010号）
