[Amplification and expression of human Artemin cDNA].
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 oxidoreduction system, and the recombinant protein is analyzed by Western blotting. Human Artemin cDNA was amplified and the recombinant protein was expressed in vitro successfully.
Cui
Yu
Y
College of Agriculture, Hainan University, Haikou 570228, China. yucui9@126.com
Li
Jing-Peng
JP
D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D009419:Nerve Tissue Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
C490886:ARTN protein, human; D018076:DNA, Complementary; D009419:Nerve Tissue Proteins