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运用脉冲场凝胶电泳进行宏观限制性分析对藤黄微球菌菌株基因组异质性的研究。

Studies on the genomic heterogeneity of Micrococcus luteus strains by macro-restriction analysis using pulsed-field gel electrophoresis.

作者信息

Murayama O, Matsuda M, Moore J E

机构信息

Laboratory of Molecular Biology, The Graduate School of Environmental Health Sciences, Azabu University, 1-17-71 Fuchinobe, Sagamihara 229-8501, Japan.

出版信息

J Basic Microbiol. 2003;43(4):337-40. doi: 10.1002/jobm.200390036.

Abstract

Macro-restriction analysis by means of double digestion using DraI and VspI demonstrated that they cleaved the genomic DNAs from Micrococcus luteus JCM1464(T), JCM3347, and JCM3348 into four to five fragments in a distinguishable manner by pulsed-field gel electrophoresis (PFGE). Separate digestion with DraI and VspI cleaved the genomic DNA from M. luteus ATCC9341 into a relatively limited number of restriction fragments (six pieces). SspI and XbaI cleaved the genomic DNA from each of the four strains into restriction fragments in a distinctly different and distinguishable manner.Thus, PFGE profiles after digestion with these four restriction enzymes that recognize six specific base sequences demonstrated the heterogeneity at the whole genomic DNA level among the four strains of M. luteus. The size of the genomic DNA of M. luteus ATCC9341 was estimated to be approximately 2.3 Mb by the summing the lengths of the restriction fragments generated by each three restriction enzymes (DraI, SspI, and VspI) and averaging the results.

摘要

通过使用DraI和VspI进行双酶切的宏观限制性分析表明,它们通过脉冲场凝胶电泳(PFGE)以可区分的方式将藤黄微球菌JCM1464(T)、JCM3347和JCM3348的基因组DNA切割成四到五个片段。用DraI和VspI分别酶切将藤黄微球菌ATCC9341的基因组DNA切割成相对有限数量的限制性片段(六个片段)。SspI和XbaI以明显不同且可区分的方式将四种菌株的基因组DNA切割成限制性片段。因此,用这四种识别六个特定碱基序列的限制性酶消化后的PFGE图谱显示了藤黄微球菌的四个菌株在全基因组DNA水平上的异质性。通过将由三种限制性酶(DraI、SspI和VspI)各自产生的限制性片段的长度相加并求平均值,估计藤黄微球菌ATCC9341的基因组DNA大小约为2.3 Mb。

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