Molecular Cloning, Sequencing, and Expression inEscherichia coliof Human Preprourokinase cDNA
| Autor: | Alfredo Cravador, F. Brockly, A Van Elsen, P Jacobs, Andrew W. Bollen, B. Colau, P. Chuchana, Albert Herzog, R. Loriau |
|---|---|
| Rok vydání: | 1985 |
| Předmět: |
Clone (cell biology)
Biology Molecular cloning Biochemistry Valine Complementary DNA Escherichia coli Genetics Humans Cloning Molecular Molecular Biology Peptide sequence Alanine chemistry.chemical_classification Enzyme Precursors Base Sequence cDNA library DNA Urokinase-Type Plasminogen Activator Molecular biology Amino acid Molecular Weight Gene Expression Regulation chemistry Protein Processing Post-Translational Plasmids |
| Zdroj: | DNA. 4:139-146 |
| ISSN: | 0198-0238 |
| DOI: | 10.1089/dna.1985.4.139 |
| Popis: | A 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. |
| Databáze: | OpenAIRE |
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