中华耳科学杂志2011，Vol.9Issue(1)：77-81,5.
野生型SLC26A4基因表达载体的构建及在HEK293细胞中的表达
The construction of wild-type SLC26A4 gene expression vector and the expression in HEK293 cells
摘要
Abstract
Objective To construct a eukaryotic expression vector containing the wild-type SLC26A4 gene and green fluorescent protein gene for clinical deafness diagnosis and prenatal diagnosis about SLC26A4-related deafness. Methods Using the Gene recombination and restriction enzyme digestion and gene sequencing, to construct and identify the eukaryotic expression plasmid of pECFP-SLC26A4. Using fluorescence microscope to observe the expression of trans-fected HEK293 cells by liposome, using reverse transcription-polymerase chain reaction (RT-PCR) to detect the mRNA, and using Western Blot to detect SLC26A4 protein expression. Results The positive recombinant gene contain the SLC26A4 fragment, and the sequence were the same sequence as on the GenBank. After being transfected Into HEK293 cells, The green fluorescent can be observed, and The SLC26A4 bands can be amplified. The 87kDa -sized protein was detected by Western Blot. Conclusion we successfully constructed the eukaryotic expression vector containing wild-type SLC26A4 and EGFP gene and the vector can express in mammalian HEK293 cell lines.关键词
SLC26A4/质粒/表达Key words
SLC26A4/ Plasmid/ Expression分类
医药卫生引用本文复制引用
武健,张国正,杨仕明,戴朴,袁永一..野生型SLC26A4基因表达载体的构建及在HEK293细胞中的表达[J].中华耳科学杂志,2011,9(1):77-81,5.基金项目
国家自然科学基金青年项目(30801285) （30801285）
北京市科技新星计划(2009B34)资助. （2009B34）
