Optimal conditions for detection of reverse transcriptase activity in human placentas.
Optimal conditions for detecting reverse transcriptase activity in human placental extracts are described. They vary with the state of the placenta at birth and are influenced by relative amounts of detergent, monovalent cation, and protein in the reaction mixture. Demonstrating activity of the placental enzyme requires detergent, but the enzyme is sensitive to high detergent concentrations. This sensitivity can be be altered by lowering the monovalent cation concentration from 0.154 to 0.034 M and by adding protein to the reaction mixture. The detection of reverse transcriptase in Rauscher murine leukemia virus and baboon endogenous type C virus, but not in the Mason-Pfizer monkey type D virus, shows similar requirements.
D000818:Animals; D002414:Cations, Monovalent; D003902:Detergents / Q000494:pharmacology; D005260:Female; D006801:Humans; D007700:Kinetics; D010215:Papio; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D011925:Rauscher Virus / Q000201:enzymology; D012190:Retroviridae / Q000201:enzymology; D013045:Species Specificity
D002414:Cations, Monovalent; D003902:Detergents; D012194:RNA-Directed DNA Polymerase