Department of Botany, University of Durham, South Road, DH1 3LE, Durham, U.K..
Plant Mol Biol. 1984 Mar;3(2):73-81. doi: 10.1007/BF00040031.
Two oligonucleotide sequences were synthesised by a solid-phase phosphotriester method. One of these sequences, A was a copy of part of a characterised cDNA clone encoding the basic subunit of legumin, a seed storage protein of Pisum sativum L. (garden pea); the other sequence B was predicted to be complementary to the 5' region of legumin mRNA on the basis of the amino acid sequence of legumin acidic subunits and most likely codon usage. Sequence A was shown to hybridise specifically to a legumin cDNA clone and to legumin mRNA. Sequence B did not hybridise specifically to legumin mRNA and was concluded not to be correctly complementary to legumin mRNA. Sequence A was used as a primer for cDNA synthesis using pea seed mRNA as a template. The cDNA so produced hybridised specifically to a legumin cDNA clone, to legumin mRNA, and to sequences encoding legumin in a restriction digest of pea genomic DNA. It is suggested that such oligonucleotide primed cDNAs may be of general value in probing eukaryotic genomic DNA.
两个寡核苷酸序列通过固相磷酸三酯法合成。其中一个序列 A 是豌豆贮藏蛋白豆球蛋白碱性亚基编码部分 cDNA 克隆的拷贝；另一个序列 B 根据豆球蛋白酸性亚基的氨基酸序列和最可能的密码子使用情况，预测是豆球蛋白 mRNA 5' 区的互补序列。序列 A 特异性杂交到豆球蛋白 cDNA 克隆和豆球蛋白 mRNA。序列 B 没有特异性杂交到豆球蛋白 mRNA，因此推断它与豆球蛋白 mRNA 不互补。序列 A 用作豌豆种子 mRNA 为模板的 cDNA 合成的引物。所产生的 cDNA 特异性杂交到豆球蛋白 cDNA 克隆、豆球蛋白 mRNA 以及豌豆基因组 DNA 限制酶切物中编码豆球蛋白的序列。建议此类寡核苷酸引发的 cDNA 可能在探测真核基因组 DNA 方面具有普遍价值。
