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

FEBS Letters

Godovikova, T.S.¹  Koshkin, A.A.¹  Lavrik, O.I.¹  Frolova, E.I.¹  Zarytova, V.F.¹  Lokhova, I.A.¹  Kavsan, V.M.³  Khomov, V.V.²  Rudenko, N.K.²  Sergeev, D.S.¹  Nevinsky, G.A.¹  Ivanova, E.M.¹ 

[1] Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Academy of Sciences of the USSR, Novosibirsk 630090, USSR;Scientific Research Institute of Design and Technology of Biologically Active Substances, Berdsk, Novosibirsk region 633190, USSR;Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR, Klev 252627, USSR

关键词: Klenow fragment;    AMV reverse transcriptase;    5′-Derivatives d(pT)8;    Affinity;    FK;    Klenow fragment of E. coli DNA polymerase 1;    AMV-RT;    reverse transcriptase from avian myeloblastosis virus;    Et-Phn-;    Hem-;    Chs-;    Dnm-;    derivatives of ethidium;    N-(2-hydroxyethyl)-phenazinium;    hemin;    cholesterol and daunomycin in d(pT)8 analogs;    respectively;   

DOI  :  10.1016/0014-5793(91)80371-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The K _m and V _max 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 K _m 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 V _max 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.

【 授权许可】

Unknown