N.BspD6I DNA nickase strongly stimulates template-independent synthesis of non-palindromic repetitive DNA by Bst DNA polymerase
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Abstract
Highly efficient DNA synthesis without template and primer DNAs occurs when N.BspD6I DNA nickase is added to a reaction mixture containing deoxynucleoside triphosphates and the large fragment of Bst DNA polymerase. Over a period of 2 h, virtually all the deoxynucleoside triphosphates (dNTPs) become incorporated into DNA. Inactivation of N.BspD6I nickase by heating inhibits DNA synthesis. Optimal N.BspD6I activity is required to achieve high yields of synthesized DNA. Electron microscopy data revealed that the majority of DNA molecules have a branched structure. Cloning and sequencing of the fragments synthesized demonstrated that the DNA product mainly consists of multiple hexanucleotide non-palindromic tandem repeats containing nickase recognition sites. A possible mechanism is discussed that addresses template-independent DNA synthesis stimulated by N.BspD6I nickase.
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Articles in the same Issue
- Localization and organization of protein factors involved in chromosome inheritance in Dictyostelium discoideum
- N.BspD6I DNA nickase strongly stimulates template-independent synthesis of non-palindromic repetitive DNA by Bst DNA polymerase
- Glutamic acid-141: a heme ‘bodyguard’ in anionic tobacco peroxidase
- SUMOylation modulates transcriptional repression by TRPS1
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- Insulin-regulated aminopeptidase: analysis of peptide substrate and inhibitor binding to the catalytic domain
- Obesity resistance of the stearoyl-CoA desaturase-deficient (scd1-/-) mouse results from disruption of the epidermal lipid barrier and adaptive thermoregulation
- Characterisation of novel splicing variants of the tyrosine hydroxylase C-terminal domain in human neuroblastic tumours
- Enzymatic properties of human kallikrein-related peptidase 12 (KLK12)
- Down-regulation of human extracellular cysteine protease inhibitors by the secreted staphylococcal cysteine proteases, staphopain A and B
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