摘要
用0.3M的甘氨酸缓冲液（含0.1M的MgCl<sub>2</sub>,pH7.4）对泡桐丛枝病病树的新鲜或冰冻组织进行抽提,用蜗牛酶处理,并差速离心,获得泡桐丛枝病类菌原体;然后用蛋白酶E处理,苯酚和氯仿抽提,酒精沉淀,可有效地制备得泡桐丛枝病类菌原体总核酸。所得总核酸在0.6％的琼脂糖凝胶电泳时呈现出大分子和小分子两种电泳带。DNase I RNase A及专一性降解单链核酸的S.酶等分析结果表明,大分子带为DNA,小分子带为RNA,且DNA具有双链性质,RNA具有单链性质。将上述总核酸用RNase A消化,苯酚和氯仿抽提,乙醇沉淀,可获得单一的泡桐丛枝病类菌原体DNA。电泳测得其DNA分子量约为1.5×10<sup>7</sup>道尔顿。目前国内外均未见泡桐丛枝病类菌原体核酸报导。
The paulownia witches' broom mycoplasmas like organism (MLD) was separated from the leaves of the diseased tree by treatment with qlycine-buffer, snailase digestion and differential centrifugation. The MLD nucleic acid was prepared by protease E digestion,phenol-chloroform extract and ethanol precipitate. Then, the MLD nucleic acid was separated to two kinds of different component on 6.6% agarose gel electrophoresis. DNase I, RNase A and S_1 engyme analysis proved that the bigger molecular is DNA is about 1.5×10~7 dalton. It hasn't been reported about the research of the nucleic acid of paulownia witches' broom MLD at home and abroad.
出处
《微生物学杂志》
CAS
CSCD
1990年第3期34-37,共4页
Journal of Microbiology
