The affinity of the Klenow fragment of E. coli DNA‐polymerase 1 to primers containing bases noncomplementary to the template and hairpin‐like elements

FEBS Letters

Nevinsky, Georgy A.¹  Levina, Asja S.¹  Kolocheva, Tatjana I.¹  Ljach, Marija V.¹  Gorn, Vladimir V.¹ 

[1] Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the USSR Academy of Sciences, Novosibirsk 630090 USSR

关键词: Klenow fragment;    Primers;    Abnormally high affinity;    FK;    Klenow fragment of E. coli DNA-polymerase 1;    V 0;    V max;    initial and maximum rates of polymerization reaction;    respectively;   

DOI  :  10.1016/0014-5793(92)80155-A

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 a set of primers: d(pT)_n (pC)(pT)_m (n=3–9.m=0–7) and d(pT)₄ (pCpG)₄ (pT)₄ (k=1–5) have been estimated. Poly(dA) was used as template. The number of complementary bases from the 3′ end to a noncomplementatry ones was shown to determine the efficiency of interaction of d(pT)_n (pC) (pT)_m with the Klenow fragment. Oligonucleotides d(pT)₄ (pCpG)₄ (pT)₄ in solution forming duplexes containing hairpin-like elements, show a higher affinity to the enzyme that control d(pT)₄, d(pT)_n, and d(pT)_n (pC)(pT)_m primers. For example, the K _m value (1.1 nM) for d(pT)₄ (pCpG)₅ (pT)₄ is about 14,000 and 200 times lower than those for d(pT)₄ and d(pT)_? respectively. Possible reasons for such an abnormally high affinity of the above primers are discussed.

【 授权许可】

Unknown