Cloning and Expression in Escherichia coli of Full-Length Complementary DNA Coding for Human alpha1-Antitrypsin
Bollen, A., Herzog, A., Cravador, A., Herion, P., Chuchana, P., Van Der Straten, A., Loriau, R., Jacobs, P., & 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. https://doi.org/10.1089/dna.1983.2.255
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.