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Molecular cloning, sequencing, and expression in Escherichia coli of human preprourokinase cDNA

1985Journal article Restricted access
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dc.contributor.authorJacobs, P.
dc.contributor.authorCravador, A.
dc.contributor.authorLoriau, R.
dc.contributor.authorBrockly, F.
dc.contributor.authorColau, B.
dc.contributor.authorChuchana, P.
dc.contributor.authorVan Elsen, A.
dc.contributor.authorHerzog, A.
dc.contributor.authorBollen, A.
dc.date.accessioned2015-06-15T09:15:56Z
dc.date.available2015-06-15T09:15:56Z
dc.date.issued1985
dc.description.abstractA cDNA library derived from human carcinoma cells was used to isolate a clone, pULB1000, coding for the preproenzyme form of human urokinase. This clone carries the full-length sequence coding for the signal peptide and for the A chain (157 amino acids) and B chain (253 amino acids) of urokinase in tandem. The sequence of the cDNA predicts the presence of a single lysine residue between the last amino acid of the mature A polypeptide (Phe-157) and the first amino acid of the mature B polypeptide (Ile-1). The amino acid sequence deduced from the cDNA sequence fits the published amino acid sequence data with three exceptions, the reported cysteine residue at position 131 in the A chain is a tryptophan, and glycine 366 and alanine 410 in the B chain are, respectively, a cysteine and a valine in our clone. A large Bgl I fragment (1482 bp), derived from the clone pULB1000 coding for most of the signal peptide and for the A and B chains, has been subcloned into the expression vector pCQV2. Heat induction of E. coli cells carrying the recombinant plasmid leads to the production of urokinase-like polypeptides having the expected molecular weights and being specifically recognized by antibodies raised against natural human urokinase.
dc.identifier.issn0198-0238
dc.identifier.otherAUT: ACR00659;
dc.identifier.urihttp://hdl.handle.net/10400.1/6163
dc.language.isoeng
dc.peerreviewedyes
dc.relation.isbasedonP-008-R6H
dc.titleMolecular cloning, sequencing, and expression in Escherichia coli of human preprourokinase cDNA
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage146
oaire.citation.startPage139
oaire.citation.titleDNA
oaire.citation.volume4
person.familyNameCravador
person.givenNameAlfredo
person.identifier.orcid0000-0002-9831-9815
person.identifier.ridK-7247-2012
person.identifier.scopus-author-id6602537257
rcaap.rightsrestrictedAccess
rcaap.typearticle
relation.isAuthorOfPublication8c84a9e6-6b7f-4b50-93a2-152c85901722
relation.isAuthorOfPublication.latestForDiscovery8c84a9e6-6b7f-4b50-93a2-152c85901722

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