Koller B, Delius H, Dyer T A
Eur J Biochem. 1982 Feb;122(1):17-23. doi: 10.1111/j.1432-1033.1982.tb05842.x.
The DNAs of the plasmids pTac39 and pZmB1B, which contain the wheat chloroplast DNA BamHI fragment 2 and the maize chloroplast DNA BamHI fragment 9 respectively, were analysed by electron microscopy and by Northern blotting. Both plasmids contain the gene for the large subunit of ribulose bisphosphate carboxylase (LS gene). Comparison of the DNAs of the two species by partial denaturation and heteroduplex analysis shows extended homologies interspersed with a fine pattern of non-homology. On analysis of wheat-RNA X maize-DNA hybrids a region of non-homology at the 3' end of the LS mRNA of about 200 bases was found. Analysis of wheat DNA X RNA hybrids showed that the wheat LS mRNA contains 1700 +/- 50 bases and another hybrid, containing 2400 bases was also shown to occur in a position adjacent to that formed by the LS mRNA. Binding of Escherichia coli RNA polymerase and the start of transcription in vitro occurred at well-defined sites, which are located between or close to the positions where mRNA X DNA hybrids are found. A modified technique for the preparation and fractionation by electrophoresis of glyoxalated RNA is described.
分别含有小麦叶绿体DNA BamHI片段2和玉米叶绿体DNA BamHI片段9的质粒pTac39和pZmB1B的DNA,通过电子显微镜和Northern印迹法进行了分析。两种质粒都含有核酮糖二磷酸羧化酶大亚基的基因(LS基因)。通过部分变性和异源双链分析对两种物种的DNA进行比较,结果显示存在延伸的同源性,其间穿插着精细的非同源模式。在分析小麦RNA与玉米DNA杂交体时,发现在LS mRNA 3'端约200个碱基处存在一个非同源区域。对小麦DNA与RNA杂交体的分析表明,小麦LS mRNA含有1700±50个碱基,并且还显示出另一种含有2400个碱基的杂交体出现在与LS mRNA形成的位置相邻的位置。大肠杆菌RNA聚合酶的结合及体外转录起始发生在明确的位点,这些位点位于发现mRNA与DNA杂交体的位置之间或附近。本文描述了一种用于制备经乙二醛化的RNA并通过电泳进行分离的改良技术。