CLONING, SEQUENCE AND EXPRESSION OF THE PEL GENE FROM AN AMYCOLATA SP.

摘要: Abstract The pel gene from an Amycolata sp. encoding a pectate lyase (EC 4.2.2.2) was isolated by activity screening genomic DNA library in Streptomyces lividans TK24. Subsequent subcloning and sequencing of 2.3 kb Bam HI– Bgl II fragment revealed open reading frame 930 nt corresponding to protein 29 660 Da. overall G+C content for the coding region 65%, with strong preference third (wobble) codon position (93%). A putative ribosome-binding site 5′-GGGAG-3′ preceded translational start 7 base pairs. contains signal peptide 26 amino acids, that is cleaved after sequence Ala–Thr–Ala. size deduced as well its N-terminal amino-acid match wild-type Expression S. TK24 resulted high culture supernatant, concomitant appearance dominant band on sodium dodecyl polyacrylamide gel at 30 kDa. No detected E. coli BL21 under T7 promotor. showed 40% identity PelE Erwinia chrysanthemi Glomerella cingulata . clearly belongs superfamily, sharing all functional acids likely has similar structural topology Pels Bacillus subtilis