The effect of single-stranded DNA binding proteins on template/primer-independent DNA synthesis in the presence of nicking endonuclease Nt.BspD6I |
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| Authors: | N V Zyrina R I Artyukh I V Svad’bina L A Zheleznaya N I Matvienko |
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| Institution: | 1.Institute of Theoretical and Experimental Biophysics,Russian Academy of Sciences,Pushchino, Moscow oblast,Russia;2.Institute of Protein Research,Russian Academy of Sciences,Moscow,Russia |
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| Abstract: | Nt.BspD6I nicking endonuclease stimulates template/primer-independent DNA synthesis by Bst DNA polymerase. Template/primer-independent
DNA synthesis may be one of the reasons for the formation of nonspecific products in certain DNA amplification reactions,
especially those involving nicking endonucleases. Expansion of the range of DNA amplification procedures performed in the
presence of nicking endonucleases makes the search for template/primer-independent DNA synthesis inhibitors highly relevant.
The present work has shown that a single-strand DNA binding protein from E. coli does not affect template/primer-independent DNA synthesis regardless of the presence or absence of Nt.BspD6I. A single-stranded
DNA-binding protein coded by gene 32 from bacteriophage T4 completely inhibits template/primer-independent DNA synthesis in the absence of nicking endonuclease.
If nicking endonuclease is present, the protein does not suppress the synthesis of the specific product but causes a significant
decrease of the amount of template/primer-independent DNA synthesis products. |
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