[Studies of cloning and expression of rat alpha-amidase gene in Streptomyces lividans].
alpha-Amidase catalyzes the C-terminal amidation of active polypeptides in the Nerve-Endocrine system. It is important for full biological activity of the polypeptides. By using the total RNA of rat atrium as a templet, the cDNA encoding alpha-amidase was amplified by RT-PCR and sequenced. The cDNA was fused with the coding sequence of mel C1 signal peptide to produce fusion gene mel/AE for secretive expression in Streptomyces lividans. The fusion gene mel/AE was inserted into plasmid pIJ680 and the recombinant plasmid pIJ-mel/AE680 was obtained. The results of SDS-PAGE and biological activity showed that the recombinant strain S. lividans TK54[pIJ-mel/AE 680] produced secretly alpha-amidase.
Wu
B Y
BY
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Peking Medical College, Beijing 100050.
Li
Y
Y
D000818:Animals; D003001:Cloning, Molecular; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013302:Streptomyces / Q000235:genetics*; D000516:alpha-Amylases / Q000235:genetics*
D011994:Recombinant Proteins; D000516:alpha-Amylases