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油菜细胞质雄性不育系叶绿体DNA特异片段的分子克隆
引用本文:高洁,孔繁瑞,李继耕.油菜细胞质雄性不育系叶绿体DNA特异片段的分子克隆[J].遗传学报,1987(5).
作者姓名:高洁  孔繁瑞  李继耕
作者单位:中国科学院遗传研究所 北京 (高洁,孔繁瑞),中国科学院遗传研究所 北京(李继耕)
摘    要:采用高离子浓度缓冲液法分别提取油菜不育系及保持系的叶绿体DNA。经Sepharcse 4B柱层析纯化后,得到具有较高纯度的叶绿体DNA样品。将其分别用Eco RI、Bam HI、HimdHI、PstI和XhoI 5种限制性内切酶酶解,得到5种限制性内切酶图谱。其中除PstI图谱外,其它4种酶谱均显示出明显的两系间叶绿体DNA结构差异。以pBR 322为载体,分别克隆了不育系Bam HI图谱上的3个特异片段。得到的3个克隆,经克隆杂交及电泳分析后,证实分别带有上述3个目的片段。这些特异片段的特性及其与花粉育性的关系尚在研究中。

关 键 词:雄性不育性  分子克隆  叶绿体DNA

Molecular Cloning of Specific Fragments of ctDNA from Male-sterile Line of Rape
Gao Jie Kong Fanrui Li Jigeng.Molecular Cloning of Specific Fragments of ctDNA from Male-sterile Line of Rape[J].Journal of Genetics and Genomics,1987(5).
Authors:Gao Jie Kong Fanrui Li Jigeng
Abstract:The ctDNAs of a male-sterile line from a rape variety with radish cytoplasm and its fertile counterpart were isolated.Having been purified through Sepharose 4B gel chromatography,the CtDNAs were digested with EcoRI,Bamii],HindIII,Pst1 and XhoI,and the restriction patterns were obtained.Of all the patterns,there are distinct differences between the male sterile line and its counterpart except Pst1 pattern.The DNA of three specific fragments from BamHI pattern of male-sterile line was recovered.After incubated with plasmid pBR322 digested by BamHl as well as T4-DNAligase respectively,three recombinants were obtained.Then the re-combinants were used to transform E.coli HB101.Through resistant screening,clone hybridization and electrophoresis analysis,three clones earring the specific fragments respeccively were got.Now the characters of these fragments are being investigated.
Keywords:Male-sterile  Molecular cloning  Chloroplast DNA
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