Construction and expression of prokaryotic vector of a novel candidate gene IDD01 associated with hepatocellular carcinoma
- VernacularTitle:肝细胞癌候选相关基因IDD01原核表达载体的构建与表达
- Author:
Xiangyu CHEN
;
Fangling DUAN
;
Jiansheng LI
;
Jun MA
;
Lexun XUE
;
- Publication Type:Journal Article
- Keywords:
HCC;
IDD01 gene;
fusion expression;
SDS PAGE
- From:Journal of Third Military Medical University
2003;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the prokaryotic vector and to express a novel candidate gene IDD01 associated with hepatocellular carcinoma (HCC) for its functional study. Methods The open reading frame (ORF) of IDD01, amplified from HCC tissue by RT PCR, was inserted into the expression vector pMAL c2X. The recombinant vector was transformed into E. coli TB1, and the expression products were analyzed by SDS PAGE. Results The length of PCR product was about 770 bp. The restriction enzyme digestion and sequencing conformed that the gene segment was correctly inserted into the vector. SDS PAGE and density scanning indicated that the protein was expressed as a fusion protein with 28 KD of molecular weight and the fusion protein possessed 30.4% of the total bacterial protein. Conclusion The recombinant vector is constructed successfully and IDD01 can be expressed at a high level, which lays a foundation for the further research on the functions of IDD01 gene.
