title: Synthesis and expression of a gene from kringle-2 domain of tissue plasminogen activator in E. coli.
abstract: The DNA fragment corresponding to the tissue plasminogen activator (tPA) sequence 174-262 (Kringle-2 domain) has been synthesized by using the solid phase phosphotriester method. The Kringle-2 domain of human tPA was expressed in Escherichia coli by secretion into the periplasmic space using the Lpp-Lac promoter and PIN-III OmpA2 signal sequence. About two thirds of the expression product was secreted into the periplasmic space, and purified with ammonium sulfate fractionation, affinity chromatography on Lysine-Sepharose, and FPLC-Mono Q exchange chromatography. The amino acid composition observed from the Kringle-2 purified from E. coli is identical with that expected for the 174-262 fragment of human tPA. Radio binding assay shows that the recombinant Kringle-2 domain possesses the activity of fibrin binding.
authors.affiliation: Department of Biochemistry, Nanjing University, PRC.
authors.forename: Z C
authors.initials: ZC
authors.lastname: Hua
authors.forename: H L
authors.initials: HL
authors.lastname: Fu
authors.forename: Y H
authors.initials: YH
authors.lastname: Chen
authors.forename: R R
authors.initials: RR
authors.lastname: Yu
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: D X
authors.initials: DX
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D018082:Kringles / Q000235:genetics*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis; D010959:Tissue Plasminogen Activator / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D010446:Peptide Fragments; D004247:DNA; D010959:Tissue Plasminogen Activator