关键词: Artemin, RT-PCR, 原核表达

Abstract:

The cDNA fragment encoding human Artemin was amplified by RT-PCR with human fetal brain RNA as template. Result of sequencing showed that the homology of nucleotides is 99.7% between the amplified human Artemin cDNA and the reported one (GenBank accession No. AF115765) and the homology of amino acids is 100%. The prokaryotic expressing vector pGEX-6p-1-hART carrying the amplified DNA fragment was obtained by ligation with the plasmid pGEX-6p-1. The expression of recombinant human Artemin fusion protein in E.coli was analyzed by SDS-polyacarylamide gel electrophoresis. Result demonstrated that the recombinant protein, mostly found in inclusion bodies, accounted for 18.32% of the total bacterial lysate. The inclusion body was dissolved and renatured by the oxidoreducuion system, and the recombinant protein is analyzed by Western blotting. Human Artemin cDNA was amplified and the recombinant protein was expressed in vitro successfully.