PMID: 2493504Mar 15, 1989Paper

cDNA cloning and expression of human complement component C2

The Journal of Immunology : Official Journal of the American Association of Immunologists
T HoriuchiJ E Volanakis

Abstract

A full-length cDNA clone for C component C2 was isolated from a human liver cDNA library in lambda gt11 by initially screening with an affinity-purified rabbit anti-C2 antibody and then using the isolated partial C2 cDNA as a probe for re-screening the library. The cDNA insert of clone lambda C2HL5-3 was sequenced in its entirety. It consisted of 2961 nucleotides including a 5' untranslated region of 388 nucleotides, followed by a 60 nucleotide region coding for a putative signal peptide, a 2196 nucleotide region coding for the 732 amino acids of the mature C2 polypeptide, and a 317 nucleotide long 3' untranslated region. Comparison of the nucleotide sequence to the previously reported C2 cDNA sequence showed two major differences. First, the 5' untranslated region of C2HL5-3 was 352 nucleotides longer and included four ATG followed by in-frame termination codons. Second, nucleotide residue 1987 was a C instead of a G, resulting in a change of amino acid residue 513 from Leu to Phe and in the appearance of an EcoRI site. The full-length C2 cDNA was cloned into the expression vector p91023(B). Transfection of the recombinant plasmid in COS cells resulted in the secretion of a protein with antigenicity and hemolytic activity simi...Continue Reading

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