Abstract
The influence of factors on the substrate-specificity ofPst I restriction endonuclease has been studied with the method of electrophoresis. The results show that, the specificity ofPst I almost can not be influenced by the single alteration of the concentration of Tris·HCl, Mg2+ or Na+ in the reaction system, but it can be altered by the reduction of any two of them. The specificity can not be altered by the single alteration of pH or the replacement of Mg2+ with Mn2+. The addition of glycerol or dimethylsulphoxide (DM-SO) to the reaction system results in the relaxation of the substrate-specificity ofPst I, but dimethyl-methylformide, glycol and ethyl alcohol can not bring about the alteration ofPst I specificity. Through the method of cloning and sequencing, the nucleotides of No. 1 and 6 in the recognition sequence ofPst I have changed (1C→A or 6G→T). Used with the enzyme analysis of an artificially synthetic DNA segment containing a special sequence, the nucleotides of No. 1 and 6 have both changed (1C→A and 6G→T). The recognition sequence ofPst I is speculated to be changed from CTGCA→G to TGCA→.
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Foundation item: Supported by the Research Fund for the Doctoral Program of Higher Education.
Biography: Zou Guo-lin (1947-), male, professor. research direction, biochemistry.
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Guo-lin, Z., Cheng-zhuo, G., Xin-chun, P. et al. Alteration of the specificity ofPst I restriction endonuclease. Wuhan Univ. J. Nat. Sci. 5, 361–365 (2000). https://doi.org/10.1007/BF02830157
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DOI: https://doi.org/10.1007/BF02830157