cDNA cloning of human proglucagon from brain tissue

Objective To construct the recombinant eukaryotic plasmid of human proglucagon(PG) and to prepare for expressing in eukaryotic cells.Methods Total RNA as well as mRNA were isolated from human brain tissue and cDNA encoding PG was amplified by reverse transcription PCR(RT-PCR).The cDNA was inserted into the vector pVITRO3.The constructed vector pVITRO3-PG cDNA was identified by restriction endonucleases and sequenced.Results No discrepancy was found in nucleotide as compared with standard sequence of PG from GenBank.It encoded 229 amino acids,whose length was 687 bp.Conclusion The cDNA of PG is successfully obtained and the recombinant plasmid is constructed correctly.It will be helpful for analyzing the function of recombinant PG.