Gene cloning, expression and activity detection of porcine interleukin-18 mature protein in Escherichia coli.
- Author:
Lanlan ZHENG
1
;
Yunfei JIA
;
Baoan CUI
;
Hongying CHEN
;
Zhanyong WEI
;
Ruiliang CHEN
Author Information
1. College of Animal Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Animals;
Base Sequence;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Interleukin-18;
biosynthesis;
genetics;
Molecular Sequence Data;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
immunology;
Swine
- From:
Chinese Journal of Biotechnology
2008;24(2):214-219
- CountryChina
- Language:Chinese
-
Abstract:
Porcine interleukin-18 mature protein gene was amplified from porcine spleen cells by RT-PCR. PCR product was cloned into the T vector pGEM-T for sequencing. The nucleotide sequence of this gene was 474 bp. Then, it was subcloned into the prokaryotic expressing plasmid vector pGEX6P-1 and transformed into host E. coli strain BL21 for expression. The expression of pIL-18 mature protein gene was identified by SDS-PAGE .The expression product was fusion protein with molecular weight of 45 kD and the percentage of expression protein in E. coil protein was 28%. The protein was purified by washing of inclusion bodies and the activity was measured by methyl thiazolyl tetrazolium (MTT).
