Cloning of fibrinolytic protease-0 (<i>Efp-0</i>) gene from diverse earthworm individuals

Abstract

Fibrinolytic enzyme has potential role as therapeutic agents for the treatment of some blood clotting diseases. It is a novel oral-administered fibrinolytic agent used for the prevention and treatment of cardiac and cerebrovascular diseases. As a drug, it has been widely studied and used for clinical treatment. In the present study, we have cloned and sequenced partial <i>Eisenia fetida </i>fibrinolytic protease (<i>Efp-0</i>) gene from three earthworm strains collected from diverse habitats. The gene was amplified by using polymerase chain reaction and cloned in pGEM-T easy vector. The size of <i>Efp-0 </i>gene, cloned from the two strains of <i>E. fetida </i>and one strain of<i> Eudrilus eugeniae, </i>was 658 bp, 658 bp and 660 bp, respectively after sequencing,. Multiple sequence alignment of the cloned gene along with gene sequences available in public database showed 21 SNPs and 5 InDels in the coding region. However, diversity observed in <i>Efp-0</i> gene cloned from different earthworm strains will provide basis for allele mining of this gene. It is concluded that <i>Efp-0</i> is a novel gene and can be utilized for industrial applications. The gene sequences obtained in this study have been submitted in the GenBank (Accession no. GQ385193, GQ385194, GQ385195).