Gene cloning and expression of human chemokine ELC
- VernacularTitle:人趋化因子ELC的克隆与表达
- Author:
Fukang LUO
;
Dabin SHEN
;
Hon ZHENG
- Publication Type:Journal Article
- Keywords:
ELC;
fusion expression;
vector;
deletion mutation;
Western blotting;
protein purification
- From:Journal of Third Military Medical University
2002;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone human chemokine ELC and express the ELC fusion protein. Methods Total RNA from human inflammatory tonsil was extracted and the cDNA was generated with reverse transcription. Mature ELC gene was amplified with PCR and NcoⅠand EcoRⅠ sites were added to the 5′ and 3′ terminal respectively, and then cloned into pET32a(+). E.coli DH5? was transformed with the recombinant plasmid, and positive clones were selected. The inserted DNA was verified by enzyme digestion and DNA sequencing. The fusion expression vector of mature ELC was formed with deletion mutation. ELC expression was analyzed by SDS-PAGE and Western blotting and the ELC fusion protein was purified. Results Human chemokine ELC was successfully cloned and the fusion protein was expressed and purified. Conclusion The ELC fusion protein was expressed with solubility.
