小麦G-型细胞质雄性不育的育性恢复因子Rf3的研究Ⅱ.总体DNA构建提莫菲维小麦基因组文库

植物研究 ›› 1997, Vol. 17 ›› Issue (1): 89-95.

小麦G-型细胞质雄性不育的育性恢复因子Rf3的研究Ⅱ.总体DNA构建提莫菲维小麦基因组文库

高东杰, 曹海明, 林江南, 王同昌

哈尔滨师范大学生物系, 哈尔滨 150080

收稿日期:1996-09-01 出版日期:1997-03-15 发布日期:2016-06-14
基金资助:
国家及黑龙江省自然科学基金资助项目

STUDIES ON FERTILITY RESTORER GENE Rf3 AGAINST G-TYPE CYTOPLASMIC MALE STERILE IN WHEAT Ⅱ GENOMIC LIBRARY CONSTRUCTION OF T.TIMOPHEEVI USING TOTAL DNA

Gao Dong-jie, Cao Hai-ming, Lin Jiang-nan, Wang Tong-chang

Department of Biology, Harbin Normal University, Harbin 150080

Received:1996-09-01 Online:1997-03-15 Published:2016-06-14

摘要/Abstract

摘要： 分别用Pst Ⅰ、Hind Ⅲ和EcoR Ⅰ三种限制性内切核酸酶切割提莫菲维小麦基因组DNA,再与用相应酶处理的载体pUC119连接,然后转化到大肠杆菌菌系DH5α之中。转化效率为1.2×10⁴/μg。经α互补检测,初步筛选重组子。随机挑取单菌落分别培养,提取质粒,经琼脂糖凝胶电泳和斑点杂交,最后确定重组子,重组率为43.8%。粗略地估计了克隆片段在基因组中的拷贝数。

关键词: 提莫菲维小麦, 基因组文库, 斑点杂交, 育性恢复因子

Abstract: Genomic DNA of T.timopheevi was digested with three restriction enzymes, Pst Ⅰ, Hind Ⅲ and EcoR I.The fragments were ligated with the plasmid pUC119 which was treated with the some enzyme before.The transformation efficiency is 1.20×10⁴/μg.Recombinants were determined by means of α-complementary test, agarose gel electrophoresis and dot hybridization.The recombination frequency is 43.8%.

Key words: Fertility restorer, Genomic library, Dot hybridization, Triticum timopheevi

高东杰, 曹海明, 林江南, 王同昌. 小麦G-型细胞质雄性不育的育性恢复因子Rf3的研究Ⅱ.总体DNA构建提莫菲维小麦基因组文库[J]. 植物研究, 1997, 17(1): 89-95.

Gao Dong-jie, Cao Hai-ming, Lin Jiang-nan, Wang Tong-chang. STUDIES ON FERTILITY RESTORER GENE Rf3 AGAINST G-TYPE CYTOPLASMIC MALE STERILE IN WHEAT Ⅱ GENOMIC LIBRARY CONSTRUCTION OF T.TIMOPHEEVI USING TOTAL DNA[J]. Bulletin of Botanical Research, 1997, 17(1): 89-95.