郑州大学学报（医学版）2012，Vol.47Issue(6)：756-758,3.DOI:10.3969/j.issn.1671-6825.2012.06.003
pp GalNacT-10基因真核表达载体的构建及表达
Construction and expression of eukaryotic expression vector for pp GalNacT-10 gene
摘要
Abstract
Aim:To construct eukaryotic expression vector of pp GalNacT-10 gene. Methods:PCR synthesis full length of pp GalNacT-10 cDNA containing restriction sites( Xba I ,EcoK I ). pMD19T-GalNacT-10-ORF and pMD19T-GalNacT-10-antisense were build, and the sequence and size of pp GalNacT-10 cDNA fragment were confimed to be correct. After objective vector pcDNA3. 1 was digested,it was separately connected with GalNacT-10-ORF and GalNacT-10-antisense. Sense and antisense eukaryotic expression vectors pcDNA3. 1-GalNacT-10-ORF and pcDNA3.1-GalNacT-10-antisense were separately transfected into 293T cells. Western blot was used to identify the expressed product. Results:The results of digestion confirmed the right length of inserted DNA,which was the same as the pp GalNacT-10 cDNA,and pp GalNacT-10 protein was highly expressed in 293T cells which were transfected with pcDNA3. L-GalNacT-10-ORF,and low expressed in 293T cells which were transfected with pcDNA3. 1-GalNacT-10-antisense. Conclusion: pcDNA3. 1-GalNacT-l0-ORF and pcD-NA3.1-GalNacT-10-antisense have been successfully constructed.`关键词
pp/GalNacT-10/真核表达载体/293T细胞Key words
pp GalNacT-10/eukaryotic expression vector/293T cell分类
医药卫生引用本文复制引用
高媛,刘振华,冯菁华,李晓云,孙其喆,张宝,郑文岭,马文丽..pp GalNacT-10基因真核表达载体的构建及表达[J].郑州大学学报（医学版）,2012,47(6):756-758,3.基金项目
广东省自然科学基金资助项目 S2011040003098 （）
