榨菜线粒体DNA雄性不育相关片段克隆及序列分析

研究了榨菜(Brassica juncea var．tumida Tsen and Lee)细胞质雄性不育性(CMS)与线粒体基因组的关系,以榨菜细胞质雄性不育系及其保持系线粒体DNA(mtDNA)为模板,设计合成引物进行PCR扩增,从榨菜胞质雄性不育系mtDNA上扩增得到与雄性不育相关的特异片段T1170,点杂交分析表明,T1170仅与榨菜胞质雄性不育系mtDNA有杂交信号,而保持系的mtDNA则无,序列测定该片段全长1173bp,编码389个氨基酸,其中疏水氨基酸占35％。图4参9。

CLONING AND SEQUENCING OF A SPECIFIC MITOCHONDRIAL DNA FRAGMENT RELATED TO CYTOPLASMIC MALE STERILITY IN TUBER MUSTARD (BRASSICA JUNCEA VAR.TUMIDA TSEN AND LEE)

Based on the nucleotide sequence of "Polima"CMS-associated specific fragment orf224 in Brassica napus, two primers were synthesized and mitochondrial DNA(mtDNA) of CMS line and it's maintainer line in tuber mustard (Brassica juncea var. tumida Tsen and Lee)were used as the templates of polymerase chain reaction. An amplified fragment T 1170 about 1.2 kb was obtained only in sample of mtDNA in CMS line by PCR, which was cloned into pGEM-T vector.Dot blot indicated that a hybridized signal was detected only on mtDNA of CMS line with the probe of T 1170, which showed that T 1170 fragment was mightly correlated with CMS in tuber mustard. The length of the fragment was 1 173 base pairs and it encoded 389 amino acids including 35% ratio of hydrophobic amino acid. Fig 4, Ref 9