PURIFICATION AND GENE CLONING OF A NOVEL FIBRINOLYTIC PROTEASE FROM STREPTOMYCES SP. C3662
一种来源于链霉菌的纤溶酶的纯化及其基因的克隆

A novel fibrinolytic protease from \%Streptomyces\% sp.C3662 was purified by (NH 4) 2SO 4 fractionation,DEAE\|Sepharose and CM\|Sepharose chromatography.The molecular weight of the protease was indicated to be 30 kD by SDS\|PAGE.Using a \%E.coli/Streptomyces\% shuttle plasmid pIJ699 as the vector,shot\|gun cloning was performed to clone the gene of the protease.One clone with fibrinolytic activity harboring a plasmid that contains a DNA fragment of 6kb was obtained from 3000 clones.Sequence analysis reveals that the open reading frame of the gene of the protease is 903bp in size,encoding a putative protein of 300 amino acids with 30kD.The overall GC% and the third codon GC% of the ORF were 68 33% and 956%,respectively.Comparison of homologue showed that the purative protein is highly homologous with other proteases of Streptomyces\sp.