Expression of the Moloney murine leukemia virus and human immunodeficiency virus integration proteins in Escherichia coli.
We have constructed expression plasmids containing the genes for the MuLV and the HIV integration proteins. When introduced into Escherichia coli, these plasmids cause the production of proteins of the expected molecular weight (43K for MuLV, 31K for HIV). The wild-type MuLV coding region induces the synthesis of large amounts of integration protein; to obtain large amounts of the full-length HIV integration protein in E. coli, it was necessary to modify the coding region to disrupt a sequence that promotes efficient internal translational initiation in E. coli. It is possible to disrupt this sequence without altering the encoded amino acids; the modified plasmid makes large amounts of the full-length protein and small amounts of the internally initiated protein. Both the MuLV and HIV integration proteins require SDS to be solubilized in E. coli extracts; however, following solubilization with SDS and transfer to a nitrocellulose filter, both integration proteins bind DNA.
Hizi
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Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.
Hughes
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D003001:Cloning, Molecular; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins / Q000235:genetics*; D004926:Escherichia coli; D006678:HIV / Q000235:genetics*; D019426:Integrases; D008970:Molecular Weight; D008979:Moloney murine leukemia virus / Q000235:genetics*; D012191:Retroviridae Proteins / Q000235:genetics*; D012995:Solubility
D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D012191:Retroviridae Proteins; D004254:DNA Nucleotidyltransferases; D019426:Integrases