摘要
以小麦的rbc L为探针,分别与黑麦ctDNA EcoR I,BamH I,Hind III的酶切谱带进行Southern杂交。结果发现,E13(2.15 kb),B2(10.7 kb),H2(10.4 kb)均含有rbc L基因。用pUC9为载体克隆黑麦ctDNA的B2片段,Southern杂交后表明,得到的重组质粒含有rbc L基因,将其命名为pRCB2。设计了一种快速准确的分析方法,构建出pRCB2质粒DNA的限制性内切酶图谱,rbc L被定位在一个2.8 kb的区域内。
Wheat rbc L DNA was used as a probe to isolate the rbc L in the rye ctDNA,The BamH I-2 fragment(10·7 kb). Hind III-2 fragment (10.4 kb) and EcoR I-13fragment (2.15 kb) were found to contain the rbc L. The BamH I-2 fragment of ctDNA was ligated to BamH I site of pUC9. It wasshown by blot hybridization that the rbc L of rye had been cloned in pUC9, andthe recombinant plasmid was named pRCB2. A restriction map of pRCB2 wasconstructed by a rapid and accurate procedure based on the linearization of circularplasmid DNA in vitro by a restriction endonuclease and partial digestion of thislinearized DNA southern hybridization with ^(32)P-labeled probe. The rbc L was loca-ted in a fragment of 2.8 kb.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
1990年第4期374-379,共6页
Journal of Fudan University：Natural Science
关键词
黑麦
叶绿体DNA
无性系
酶切图谱
chloroplast DNA, cloning
Secale cereale, rbc L, restrictionmap.
