title: Molecular cloning and characterization of cDNA encoding fibrinolytic enzyme-3 from earthworm Eisenia foetida.
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.
authors.affiliation: Department of Experimental Therapy of ARS, Beijing Institute of Radiation Medicine, China.
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Dong
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Yuan
authors.forename: Ya-Jun
authors.initials: YJ
authors.lastname: Shan
authors.forename: Zhen-Hu
authors.initials: ZH
authors.lastname: Zhao
authors.forename: Jia-Pei
authors.initials: JP
authors.lastname: Chen
authors.forename: Yu-Wen
authors.initials: YW
authors.lastname: Cong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D019295:Computational Biology; D003470:Culture Media; D018076:DNA, Complementary / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005342:Fibrinolysis; D015870:Gene Expression; D002479:Inclusion Bodies / Q000378:metabolism; D008969:Molecular Sequence Data; D009835:Oligochaeta / Q000201:enzymology* / Q000235:genetics*; D016366:Open Reading Frames; D017510:Protein Folding; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
chemical_list: D003470:Culture Media; D018076:DNA, Complementary; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D010450:Endopeptidases; C083421:lumbrokinase