Llewellyn D J, Finnegan E J, Ellis J G, Dennis E S, Peacock W J
C.S.I.R.O. Division of Plant Industry, Canberra City, A.C.T., Australia.
J Mol Biol. 1987 May 5;195(1):115-23. doi: 10.1016/0022-2836(87)90331-7.
Three genomic clones for anaerobically inducible alcohol dehydrogenase (Adh) have been isolated from Pisum sativum cv. "Greenfeast" via cDNA cloning. One of these contains a complete gene, has exon sequences corresponding to one of the cDNA sequences and is likely to be an expressed gene. This gene has a structure similar to the Adh genes of maize, with introns in the same positions in the coding sequence but differing in their lengths and nucleotide sequences. At the nucleotide level the coding sequence is 75% homologous to both maize Adh1 and Adh2 and 80% homologous to the Adh gene from Arabidopsis, but has an extra coding triplet in exon 1 that is not found in the other plant Adh genes. The non-translated regions of all the gene transcripts are widely divergent between species. A short segment of the pea Adh promoter region (-290 to +57) was fused to a reporter gene and introduced into protoplasts of Nicotiana plumbaginifolia by electroporation. Transient expression of the introduced gene increased markedly when the transfected protoplasts were incubated under anaerobic conditions, showing that cis-acting regulatory signals necessary for anaerobic control of expression reside in the -290 to +57 segment. Sequence comparisons between this region and the corresponding regions of maize and Arabidopsis Adh genes have identified short sequences that may be involved in the anaerobic regulation of plant Adh genes.
通过cDNA克隆从豌豆品种“Greenfeast”中分离出了三个厌氧诱导型乙醇脱氢酶(Adh)的基因组克隆。其中一个包含完整基因,具有与其中一个cDNA序列相对应的外显子序列,可能是一个表达基因。该基因的结构与玉米的Adh基因相似,编码序列中的内含子位置相同,但长度和核苷酸序列不同。在核苷酸水平上,编码序列与玉米的Adh1和Adh2均有75%的同源性,与拟南芥的Adh基因有80%的同源性,但在第1外显子中有一个额外的编码三联体,这在其他植物的Adh基因中未发现。所有基因转录本的非翻译区在物种间差异很大。将豌豆Adh启动子区域的一小段(-290至+57)与一个报告基因融合,并通过电穿孔导入烟草叶肉原生质体。当转染的原生质体在厌氧条件下培养时,导入基因的瞬时表达显著增加,表明厌氧控制表达所需的顺式作用调节信号存在于-290至+57片段中。该区域与玉米和拟南芥Adh基因相应区域的序列比较确定了可能参与植物Adh基因厌氧调节的短序列。