摘要
经Sau3AI部分酶解并脱磷的水稻农虎26B保持系的线粒体DNA与BamHI/EcoRI双酶切的λEMBL3AB载体DNA进行了连接,对连接物进行了体外包装,并感染NM538和Q359受体菌,得到了1.75×10~4个重组子.从中随机选取5个克隆的DNA,用BamHI进行酶切,琼脂糖凝胶电泳,出现了不同于载体DNA的酶切带型.另随机选取3个克隆的DNA,均与水稻线粒体DNA发生了杂交.由此认为,水稻农虎26B保持系线粒体基因文库确已建立.
The total mitochondrial genomic DNAs of rice Nonghu 26B (Oryza Sativa) maintainer line were digested partially with Sau 3AI. After dephosphorylation with phosphatase, the resulting DNA fragments, were ligated to the vector λEMBL 3AB DNAs which had been double-digested with BamHI/EcoRI. The mixture of ligation were packaged in vitro and E. coli NM538, Q359 were infected by those packaged mixtures. 1.75×10~4 rccombinants had been, obtained. Five clones' DNAs had been chosen from them at random, and digested with BamHI. The result of agarose gel electrophoresis showed that their DNA banding patterns were obviously different from vector DNA's. In addition, three clones' DNAs were chosen at random, each of those three inserts was hybridized with the rice mtDNA probe. The results showed that the mitochondrial gene library of rice Nonghu 26B maintainer line had been constructed completely.
出处
《山东大学学报（自然科学版）》
CSCD
1991年第4期460-466,共7页
Journal of Shandong University(Natural Science Edition)
关键词
水稻
线粒体
基因文库
rice
mitochondria
gene library
