Cloning and expression in Escherichia coli of full-length complementary DNA coding for human alpha 1-antitrypsin
| Autor: | Albert Herzog, P. Chuchana, A. vander Straten, A Van Elsen, Alfredo Cravador, R. Loriau, Alex Bollen, P Herion, P Jacobs |
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| Rok vydání: | 1983 |
| Předmět: |
DNA
Bacterial congenital hereditary and neonatal diseases and abnormalities Sequence analysis DNA Recombinant Biology Biochemistry chemistry.chemical_compound Restriction map Plasmid Complementary DNA Genetics Escherichia coli Humans RNA Messenger Cloning Molecular Molecular Biology Gene Base Sequence cDNA library Proteolytic enzymes DNA DNA Restriction Enzymes Molecular biology digestive system diseases respiratory tract diseases chemistry alpha 1-Antitrypsin Plasmids |
| Zdroj: | DNA (Mary Ann Liebert, Inc.). 2(4) |
| ISSN: | 0198-0238 |
| Popis: | A cDNA library prepared from human liver was screened for alpha 1-antitrypsin, a major constituent of plasma which functions as inhibitor of proteolytic enzymes. The library was screened using a 12-base-long synthetic oligodeoxyribonucleotide corresponding to a known DNA fragment of human alpha 1-antitrypsin and by hybrid-selection of alpha 1-antitrypsin mRNA. A plasmid, pULB1523, was identified carrying a cDNA insert of about 1400 bp coding for human alpha 1-antitrypsin. Restriction mapping and DNA sequence analysis indicated that the 1400 bp code for the signal peptide and for the complete mature alpha 1-antitrypsin molecule. In addition, a solid-phase enzyme-linked immunoassay showed that pULB1523 expresses human alpha 1-antitrypsin in bacteria. Fusion of the alpha 1-antitrypsin sequence to the leader sequence of the beta-lactamase gene (plasmid pKT287) resulted also in the expression of the protein in bacteria. |
| Databáze: | OpenAIRE |
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