CONSTRUCTION AND EXPRESSION OF HUMAN Axud1 GENE WITH HA EPITOPE TAG IN LUNG ADENOCARCINOMA SPC-A1 CELL LINE
- VernacularTitle:HA表位标记的人Axud1基因的构建及在肺腺癌SPC-A1细胞中的表达
- Author:
Fan DENG
;
Zhipeng ZOU
;
Chunlu YANG
- Publication Type:Journal Article
- Keywords:
genes, Axud1;
fusion protein;
vector construction;
gene expression
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
To construct the eukaryotic vector that expresses the fusion protein of Axud1 and influenza virus hemagglutin HA epitope tag, the total RNA was isolated from the peripheral blood lymphocytes, and reverse transcription reaction was used to amplify the full length of human Axud1 cDNA. PCR product of Axud1 was then amplified using specific primers containing HA epitope sequence, and inserted into eukaryotic expression plasmid pcDNA3.1(+)digested with BamH Ⅰand Xba Ⅰ. The recombinant plasmid was identified by polymerase chain reaction, restriction endonuclease mapping and sequencing, and then transfected into human lung adenocarcinoma SPC-A1 cell lines.The fusion HA-Axud1 protein expression in anti-G418 clones was verified by Western blot. This study might be instrumental in further study of the function of Axud1 protein in tumor cells.
