5′-Derivatives of oligonucleotides as primers of DNA polymerization catalyzed by AMV reverse transcriptase and Klenow fragment of DNA polymerase 1

Academic Article

Abstract

  • The Km and Vmax values for d(pT)8 and its derivatives containing various 5′-end groups were estimated in the reaction of polymerization catalyzed with AMV-RT and FK. The change in affinity of modified primers was more pronounced in the case of AMV-RT than in the case of FK. Introducing in d(pT)8 of intercalators such as phenazinium, ethidium and daunomycin residues in 2.7-, 8.7- and 11-fold increases in the primer affinity to AMV-RT, respectively. However, in the case of hemin and cholesterol derivatives the Km values were 3 and 5 times higher than those for d(pT)8. Compared to d(pT)8, the affinity of FK to all the above analogs was 2.3-3.6 times higher with the exception of cholesterol derivative to which it was 2.4-fold lower. The effect of the 5′-end residues on the Vmax values of d(pT)8 was small and ranged from 44% to 120% of that for d(pT)8. Therefore such reactive derivatives of oligonucleotides can be used as effective primers of AMV-RT and FK. Possible reasons for various effects of the 5′-end residues of the primer on its interaction with FK or AMV-RT in the presence of poly(A) are discussed. © 1991.
  • Published In

  • FEBS Letters  Journal
  • Digital Object Identifier (doi)

    Author List

  • Lokhova IA; Nevinsky GA; Godovikova TS; Ivanova EM; Koshkin AA; Sergeev DS; Frolova EI; Rudenko NK; Khomov VV; Kavsan VM
  • Start Page

  • 111
  • End Page

  • 113
  • Volume

  • 281
  • Issue

  • 1-2