Okita T W, Cheesbrough V, Reeves C D
J Biol Chem. 1985 Jul 5;260(13):8203-13.
Near full length cDNA clones for both alpha-/beta- and gamma-type gliadins were isolated and studied for sequence diversity. Based on restriction site polymorphism and cross-hybridization studies, alpha-/beta- and gamma-type clones could be divided into five and three homology classes, respectively. Clones representing each of the different classes were sequenced and compared. Sequence divergence between the classes was due to single-base substitutions and to duplications or deletions within or near direct repeats. Thus, through numerous duplications and subsequent divergence, the gliadin multigene family encodes a polymorphic set of polypeptides differing in both isoelectric point and molecular size. Southern blot analysis of wheat DNA suggested that the number of genes encoding the alpha-/beta-type gliadins was extremely large (greater than 100 copies/haploid genome). Inasmuch as hybridization patterns were the same using DNA isolated from seeds or leaves, amplification or rearrangement of DNA does not occur during development. The complete coding sequence of a gamma-gliadin was similar to that observed for the alpha-/beta-gliadins, but with several notable differences. Comparison of gamma-type gliadin cDNA sequences showed that, unlike the conserved dodecamer repeat common to all the alpha-/beta-gliadins, the tandem repeat unit differed among gamma-gliadin clones.
分离出了α/β型和γ型麦醇溶蛋白的近全长cDNA克隆,并对其序列多样性进行了研究。基于限制性位点多态性和交叉杂交研究,α/β型和γ型克隆可分别分为五个和三个同源类。对代表每个不同类别的克隆进行了测序和比较。类之间的序列差异是由于单碱基替换以及直接重复序列内部或附近的重复或缺失。因此,通过大量的重复和随后的分化,麦醇溶蛋白多基因家族编码了一组在等电点和分子大小上都不同的多态性多肽。小麦DNA的Southern印迹分析表明,编码α/β型麦醇溶蛋白的基因数量非常多(大于100个拷贝/单倍体基因组)。由于使用从种子或叶片中分离的DNA得到的杂交模式相同,因此在发育过程中不会发生DNA的扩增或重排。γ-麦醇溶蛋白的完整编码序列与α/β-麦醇溶蛋白的相似,但有几个显著差异。γ型麦醇溶蛋白cDNA序列的比较表明,与所有α/β-麦醇溶蛋白共有的保守十二聚体重复不同,γ-麦醇溶蛋白克隆之间的串联重复单元不同。