The alpha 2 cDNA sequence of human haptoglobin carries a bacterial promoter functional in vivo.
Various constructions of human haptoglobin (Hp) cDNA coding either for the complete alpha 2FS beta precursor protein or only for the beta subunit have been placed under the control of the lambda PR promoter in the bacterial expression vector pCQV2 (Queen, 1983). In addition to the expected 45,000 dalton polypeptide synthesized after induction of the PR promoter, the complete alpha 2FS beta constructions constitutively express a smaller polypeptide of approximately 30,000 dalton corresponding to a truncated Hp protein. Computer analysis of the HpcDNA revealed the presence of two strong potential bacterial promoters (alpha 2 PF and alpha 2 PS) located in the duplicated alpha 2FS sequence. Both Hp promoter signals are followed by potential mRNA start sites and ribosome binding sites at a compatible distance from initiation codons. In addition, the Hp alpha 2 cDNA sequence, when fused upstream to the cDNA coding for alpha 1-antitrypsin, constitutively promotes in vivo the efficient expression of an hybrid protein specifically recognized by antibodies raised against alpha 1-antitrypsin or haptoglobin.
D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D006242:Haptoglobins / Q000235:genetics*; D006801:Humans; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D006242:Haptoglobins; D004247:DNA