A single 66-kilodalton polypeptide processed from the human immunodeficiency virus type 2 pol polyprotein in Escherichia coli displays reverse transcriptase activity.
We have cloned the entire pol gene of human immunodeficiency virus type 2 into a high-level Escherichia coli expression system. Induction of cultures containing the recombinant plasmid, p2RTL1, leads to rapid accumulation of polypeptides of 66, 54, and 34 kilodaltons. We have designated the larger polypeptides reverse transcriptase, and we have designated the smaller polypeptide endonuclease. Purification of reverse transcriptase via ion-exchange and affinity chromatography yields the 66-kilodalton polypeptide, with which reverse transcriptase activity is associated. Purified enzyme furthermore displays a higher apparent molecular weight than its counterpart from human immunodeficiency virus type 1.
D004926:Escherichia coli / Q000378:metabolism; D006678:HIV / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012191:Retroviridae Proteins / Q000235:genetics* / Q000378:metabolism
D010455:Peptides; D011993:Recombinant Fusion Proteins; D012191:Retroviridae Proteins; D012194:RNA-Directed DNA Polymerase