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Mammalian chromosomal DNA digestion with restriction endonucleases in silico

M.A. Abdurashitov, This email address is being protected from spambots. You need JavaScript enabled to view it. , V.A. Chernukhin, D.A. Gonchar, S.Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.2, No 3, pp 29-38, 2006

 

A theoretical method to produce digestion patterns of mammalian chromosomal DNA cleavage by restriction endonucleases was proposed. Based on recently published data on primary structures of genomes, a computer analysis was performed and diagrams of chromosomal DNA fragments distribution were plotted for DNA cleavages at 5’-GGCC-3', 5'-CCGG-3', 5'-GATC-3' and 5'-CC(A/T)GG-3' sequences. Experiments on chromosomal DNA digestion with HaeIII, MspI, Kzo9I and Bst2UI restriction endonucleases, which recognize these sites, were carried out. A correspondence between the computed diagrams and experimentally observed patterns of restriction enzymes cleavage was shown.

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An improved ARDRA method of microorganism identification and its application in identifying thermolabile alkaline phosphatase strains-producers

This email address is being protected from spambots. You need JavaScript enabled to view it. , M.A.Abdurashitov, S.Kh. Degtyarev * - corresponding author

Translated from Biotechnologia (russ.). 2005, N 6, pp 3-11

 

Based on the analysis of amplified ribosomal DNA fragmentation by restriction endonucleases, an improved method for microorganism identification (called ARDRA) has been suggested. A set of 6 restriction endonucleases (Sse9I, Tru9I, BsuRI, MspI, BstMBI and RsaI) was used to get corresponding patterns of DNA cleavage. DNA samples were isolated from four strain-producers of thermolabile alkaline phosphatase, obtained from sea water. Carrying out of ARDRA, followed by a comparison with the calculated cleavage patterns of DNA from several referenced microorganisms, allowed us to conclude that the new strains-producers belong to the Alteromonas genus.

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