Cloning and expression in Escherichia coli of full-length complementary DNA coding for human alpha 1-antitrypsin.
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Bollen A, Herzog A, Cravador A, Herion P, Chuchana P, Vander Straten A, Loriau R, Jacobs P, van Elsen A
Cloning and expression in Escherichia coli of full-length complementary DNA coding for human alpha 1-antitrypsin.
DNA. 1983;2(4):255-64.
- PubMed ID
- 6319097 [ View in PubMed]
- Abstract
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.