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Cloning and expression in Escherichia coli of full-length complementary DNA coding for human alpha 1-antitrypsin.
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.
PMID: 6319097 [PubMed - indexed for MEDLINE]
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Cited by 7 PubMed Central articles
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Cloning and molecular characterization of a human intracellular serine proteinase inhibitor.
Coughlin P, Sun J, Cerruti L, Salem HH, Bird P.
Proc Natl Acad Sci U S A. 1993 Oct 15; 90(20):9417-21.
[Proc Natl Acad Sci U S A. 1993]
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ReviewCompilation of published signal sequences.
Watson ME.
Nucleic Acids Res. 1984 Jul 11; 12(13):5145-64.
[Nucleic Acids Res. 1984]
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High-level production of biologically active human alpha 1-antitrypsin in Escherichia coli.
Courtney M, Buchwalder A, Tessier LH, Jaye M, Benavente A, Balland A, Kohli V, Lathe R, Tolstoshev P, Lecocq JP.
Proc Natl Acad Sci U S A. 1984 Feb; 81(3):669-73.
[Proc Natl Acad Sci U S A. 1984]
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