• Article

Cloning and Expression in Escherichia coli of Full-Length Complementary DNA Coding for Human alpha1-Antitrypsin

A cDNA library prepared from human liver was screened for alpha1-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 alpha1-antitrypsin and by hybrid-selection of alpha1-antitrypsin mRNA. A plasmid, pULB1523, was identified carrying a cDNA insert of about 1400 bp coding for human alpha1-antitrypsin. Restriction mapping and DNA sequence analysis indicated that the 1400 bp code for the signal peptide and for the complete mature alpha1-antitrypsin molecule. In addition, a solid-phase enzyme-linked immunoassay showed that pULB1523 expresses human alpha1-antitrypsin in bacteria. Fusion of the alpha1-antitrypsin sequence to the leader sequence of the beta-lactamase gene (plasmid pKT287) resulted also in the expression of the protein in bacteria.

Citation

Bollen, A., Herzog, A., Cravador, A., Herion, P., Chuchana, P., Van Der Straten, A., ... van Elsen, A. (1983). Cloning and Expression in Escherichia coli of Full-Length Complementary DNA Coding for Human alpha1-Antitrypsin. DNA, 2(4), 255-264. DOI: 10.1089/dna.1983.2.255

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