山东医药2011，Vol.51Issue(26)：14-16,3.
人胆固醇醋水解酶真核表达载体的构建及U937细胞系的转染
Construction of eukaryotic vector pEGFP-N1 -hCEH and establishment of stable transfectant U937 cell line
摘要
Abstract
Objective To construct human cholesterol ester hydrolase (hCEH) eukaryotic expression vector pEGFPN1 -hCEH, and observe their monocyte-macrophage cell line U937 expression. Methods Human liver cell lines L-O2 total RNA as template, amplified by RT-PCR applications hCEH coding sequence, the amplified fragments were inserted into pMD19-T vector, recovery, purification fragment was subcloned into pEGFP-N1 eukaryotic expression vector by restriction enzyme digestion, DNA sequencing, the transfected monocyte-macrophage cell line U937, and observe its expression in U937 cells. Results RT-PCR amplification of gene coding sequence hCEH obtain a 1.7 kb fragment of the treaty, in line with the expected fragment size. Recombinant plasmid pEGFP-N1-hCEH was successfully constructed with pEGFP-N1 as carrier. Gene transfection experiments showed that the recombinant express in U937 cells. Conclusions The eukaryotic expression vector pEGFP-N1-hCEH is successfully constructed, and express in U937 cells after transfection.关键词
人胆固醇酯水解酶/真核表达载体/单核巨噬细胞/U937细胞/转染Key words
cholesterol ester hydrolase/ eukaryotic expression vector/ monocyte-macrophage cell line/ U937 cell/transfection分类
医药卫生引用本文复制引用
顾立学,罗俊生,王瑞,冯旭,关宁,霍晓川..人胆固醇醋水解酶真核表达载体的构建及U937细胞系的转染[J].山东医药,2011,51(26):14-16,3.基金项目
国家自然科学基金资助项目(30971023) （30971023）
辽宁省自然科学基金资助项目(20092190) （20092190）
辽宁省教育厅高等学校科研项目(2009A481). （2009A481）
