title: [Gene cloning, expression and activity detection of porcine interleukin-18 mature protein in Escherichia coli].
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).
authors.affiliation: College of Animal Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China.
authors.forename: Lanlan
authors.initials: L
authors.lastname: Zheng
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Jia
authors.forename: Baoan
authors.initials: B
authors.lastname: Cui
authors.forename: Hongying
authors.initials: H
authors.lastname: Chen
authors.forename: Zhanyong
authors.initials: Z
authors.lastname: Wei
authors.forename: Ruiliang
authors.initials: R
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D020382:Interleukin-18 / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D013552:Swine
chemical_list: D020382:Interleukin-18; D011993:Recombinant Fusion Proteins