Ainley W M, Walker J C, Nagao R T, Key J L
Department of Botany, University of Georgia, Athens 30602.
J Biol Chem. 1988 Aug 5;263(22):10658-66.
The auxin-regulated expression of two poly(A)+ mRNAs in soybean hypocotyl was demonstrated by cloning of the cDNAs and Northern blot hybridization analyses (Walker, J.C., and Key, J.L. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 7185-7189). The corresponding genes, designated Aux28 and Aux22, have been isolated, and the cDNAs and genes have been sequenced. The Aux28 and Aux22 genes are present at one to two copies per haploid genome, contain four and two introns, and encode hydrophilic proteins of 26.8 and 21.5 kDa, respectively. Although the cDNAs were isolated independently and do not cross-hybridize under stringent hybridization conditions, the protein coding sequences of the two cDNAs have several colinear regions of high homology at the nucleic acid (77-80%) and the amino acid (80-100%) levels; together these regions constitute approximately a third of the protein coding sequences of the cDNAs. These data, together with genomic Southern blot hybridization analysis and hybrid-select translations of mRNAs homologous to the cDNAs, show that these genes belong to two related multigene families. We have identified two sequences, TGATAAAAG and GGCAGCATGCA, that occur at similar distances upstream of the transcription start site in each gene, and the spacing between these two elements is essentially identical in the two genes. The possible significance of these sequences is under evaluation.
通过cDNA克隆和Northern印迹杂交分析(Walker, J.C., and Key, J.L. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 7185 - 7189),证明了生长素调节大豆下胚轴中两种聚腺苷酸化mRNA的表达。已分离出相应的基因,分别命名为Aux28和Aux22,并对cDNA和基因进行了测序。Aux28和Aux22基因在每个单倍体基因组中以1至2个拷贝存在,分别含有4个和2个内含子,分别编码26.8 kDa和21.5 kDa的亲水性蛋白质。尽管cDNA是独立分离的,并且在严格的杂交条件下不发生交叉杂交,但两个cDNA的蛋白质编码序列在核酸(77 - 80%)和氨基酸(80 - 100%)水平上有几个高同源性的共线区域;这些区域一起约占cDNA蛋白质编码序列的三分之一。这些数据,连同基因组Southern印迹杂交分析以及与cDNA同源的mRNA的杂交选择翻译,表明这些基因属于两个相关的多基因家族。我们已经鉴定出两个序列,TGATAAAAG和GGCAGCATGCA,它们在每个基因转录起始位点上游的相似距离处出现,并且这两个元件之间的间距在两个基因中基本相同。这些序列的可能意义正在评估中。