Dolferus R, Van den Bossche D, Jacobs M
Laboratory of Plant Genetics, Vrije Universiteit Brussel, Sint-Genesius-Rode, Belgium.
Mol Gen Genet. 1990 Nov;224(2):297-302. doi: 10.1007/BF00271565.
Data presented in this paper deal with a further molecular characterization of 2 out of 32 EMS-induced Arabidopsis ADH null mutants that we isolated previously. In order to localize and characterize each mutation at the molecular level, we have cloned and completely sequenced the R002 and R006 null mutant alleles. For mutant R002, which does not contain any detectable levels of ADH protein and mRNA, we have found that the mutation is due to a single C to T base pair substitution in the reading frame; this leads to the incorporation of a TAG stop codon (amber nonsense mutation). For mutant R006, which contains normal levels of inactive protein and mRNA levels, we found a G to A base pair transition. This gives rise to a Cys to Tyr amino acid substitution in the active site of the ADH enzyme.
本文展示的数据涉及我们之前分离出的32个由甲基磺酸乙酯(EMS)诱导的拟南芥乙醇脱氢酶(ADH)缺失突变体中的2个的进一步分子特征分析。为了在分子水平上定位和表征每个突变,我们克隆了R002和R006缺失突变等位基因并进行了全序列测定。对于不含有任何可检测水平的ADH蛋白和mRNA的突变体R002,我们发现该突变是由于阅读框中单个C到T的碱基对替换所致;这导致了TAG终止密码子的插入(琥珀色无义突变)。对于含有正常水平的无活性蛋白和mRNA水平的突变体R006,我们发现了一个G到A的碱基对转换。这在ADH酶的活性位点导致了一个从半胱氨酸到酪氨酸的氨基酸替换。