Construction and identification of expressing human c-Jun N-terminal kinase(JNK)recombinant adenovirus
- VernacularTitle:表达人JNK基因重组腺病毒的构建和鉴定
- Author:
Jinhu CHEN
;
Huixia LIU
;
Jiani ZHANG
;
Min GUO
;
Yangya QUAN
;
Ying TAN
- Publication Type:Journal Article
- Keywords:
Genes,jun;
Protein kinase;
Adenoviridae/GE;
Recombination,genetic
- From:
Journal of Chinese Physician
2008;10(3):309-311
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct replication deficient recombinant adenovirus expressing human c-Jun N-terminal kinase by homologous recombination.Methods The linearized recombinant shuttle vector pAdTrack-CMV-WT-JNK was co-transformed with backbone vector pAdEasy-1 into bacteria BJ5183 for recombinant adenoviral vector.The recombinant adenoviral vector was transfected into HEK293 packing cells to construct replication deficient recombinant adenovirus,and then the recombinant adenovirus was detected by PCR and DNA sequencing.Results JNK recombinant adenoviral vector was effectively transfected into HEK 293 cells and was successfully packed by intracellular enzyme.The expression of green fluorescent protein(GFP)was observed on the 5th day after transfection.The fragment of JNK gene was amplified by PCR and identified by sequencing.The animal experiment confirmed that Ad-WT-JNK was effectivety expressed in liver tissue. Conclusion The research successfully constructed recombinant adenoviral vector and recombinant adenoviral particle.And the achievement laid a foundation for further investigation of the function and application of JNK.
