Abstract
The earthworm fibrinolytic enzyme-3 (EFE-3, GenBank accession No: AY438622), from the earthworm Eisenia foetida, is a component of earthworm fibrinolytic enzymes. In this study, cDNA encoding the EFE-3 was cloned by RT-PCR. The cDNA contained an open reading frame of 741 nucleotides, which encoded a deduced protein of 247 amino acid residues, including signal sequences. EFE-3 showed a high degree of homology to earthworm (Lumbricus rebullus) proteases F-III-1, F-III-2, and bovine trypsin. The recombinant EFE-3 was expressed in E. coli as inclusion bodies, and the gene encoding the native form of EFE-3 was expressed in COS-7 cells in the medium. Both the refolding product of inclusion bodies and the secreted protease could dissolve the artificial fibrin plate.
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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COS Cells
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Chlorocebus aethiops
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Cloning, Molecular*
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Computational Biology
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Culture Media
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DNA, Complementary / genetics*
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Electrophoresis, Polyacrylamide Gel
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Endopeptidases / chemistry*
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Endopeptidases / genetics*
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Endopeptidases / metabolism
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Escherichia coli / genetics
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Fibrinolysis
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Gene Expression
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Inclusion Bodies / metabolism
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Molecular Sequence Data
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Oligochaeta / enzymology*
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Oligochaeta / genetics*
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Open Reading Frames
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Protein Folding
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Protein Sorting Signals
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Recombinant Proteins / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Sequence Homology, Nucleic Acid
Substances
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Culture Media
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DNA, Complementary
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Protein Sorting Signals
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Recombinant Proteins
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Endopeptidases
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lumbrokinase
Associated data
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GENBANK/AY438622
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GENBANK/U25643
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GENBANK/U25648