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Медико-биологический
информационный портал
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для специалистов |
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Vol. 12, Art. 89 (pp. 1101-1118) | 2011
A new metod for preparation of highly purified the bacteriophage t4 gene 32 protein for biomedical research
N.V. Zyrina, L.A. Zheleznaya, I.V. Svadbina, N.I. Matvienko*
Russian Academy of Sciences Institute of Theoretical and Experimental Biophysics *- Russian Academy of Sciences Institute of Protein Sciences
Brief summary
Many recent advances in biomedical research can be attributed to a variety of DNA amplification methods. However the most significant problem is the appearance of nonspecifically amplified products. Employing single-stranded DNA binding (SSB) proteins from E.coli and bacteriophage T4 for DNA amplification often eliminates this problem. Thereby techniques for DNA amplification in biomedical research require the use of highly purified preparations of SSB proteins, free of DNA and nucleases. We suggest a new method for preparation of highly purified recombinant bacteriophage T4 gene 32 protein (T4gp32). Removal of nucleic acids was obtained by treatment cell lysate with spermine. Subsequent purification T4gp32 by chromatography on heparin-sepharose column allowed obtaining nuclease-free preparation better than 99% homogeneity. Key words nonspecific DNA amplification products, purification of recombinant protein T4gp32 (The article in PDF format. For preview need Adobe Acrobat Reader)  Open article in new window
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