Isolation and molecular characterization of high molecular weight glutenin subunit genes 1Bx13 and 1By16 from hexaploid wheat.
Isolation and molecular characterization of high molecular weight glutenin subunit genes 1Bx13 and 1By16 from hexaploid wheat.
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DOI：
10.1111/j.1744-7909.2007.00573.x
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发表时间：
2008-03
影响因子：
11.4
通讯作者：
B. Pang;Xueyong Zhang
中科院分区：
文献类型：
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作者：
B. Pang;Xueyong Zhang
高分子量麦谷蛋白亚基（HMW-GS）对1Bx 13 + 1By 16被认为与面包品质正相关，但其分子数据尚不清楚。为了揭示1By 16和1Bx 13高品质形成的机制，根据已发表的小麦高分子量谷蛋白基因序列设计引物，从普通小麦品种豫麦66和济麦20中扩增了1By 16和1Bx 13的开放阅读框（ORF）。1By 16的开放阅读框为2,220 bp，推导出738个氨基酸残基，含7个半胱氨酸，包括59个六肽基序和22个纳米肽基序。1Bx 13的开放阅读框为2,385 bp，由795个氨基酸组成，含4个半胱氨酸，包括68个六肽、25个纳米肽和6个三肽基序。我们发现1By 16是迄今为止在普通小麦中描述的最大的y型HMW谷蛋白基因。与1By 15相比，1By 16在中央重复结构域插入了36个氨基酸残基。细菌表达和蛋白质印迹测试证实，克隆的序列是HMW-GS 1By 16的ORF，1Bx 13是迄今为止在普通小麦中描述的最大的1Bx基因之一，与1Bx 7相比，在中央重复结构域的末端插入了六肽（PGQGQQ）。根据已发表的HMW-GS基因的全长氨基酸序列构建的系统发育树显示，1By 16与Glu-1B-2聚在一起，1Bx 13与Glu-1B-1聚在一起。
The high molecular weight glutenin subunit (HMW-GS) pair 1Bx13 + 1By16are recognized to positively correlate with bread-making quality; however, their molecular data remain unknown. In order to reveal the mechanism by which 1By16 and 1Bx13 creates high quality, their open reading frames (ORFs) were amplified from common wheat Atlas66 and Jimai 20 using primers that were designed based on published sequences of HMW glutenin genes. The ORF of 1By16 was 2,220 bp, deduced into 738 amino acid residues with seven cysteines including 59 hexapeptides and 22 nanopeptides motifs. The ORF of 1Bx13 was 2,385 bp, deduced into 795 amino acid residues with four cysteines including 68 hexapeptides, 25 nanopeptides and six tripeptides motifs. We found that 1By16 was the largest y-type HMW glutenin gene described to date in common wheat. The 1By16 had 36 amino acid residues inserted in the central repetitive domain compared with 1By15. Expression in bacteria and western-blot tests confirmed that the sequence cloned was the ORF of HMW-GS 1By16, and that 1Bx13 was one of the largest 1Bx genes that have been described so far in common wheat, exhibiting a hexapeptide (PGQGQQ) insertion in the end of central repetitive domain compared with 1Bx7. A phylogenetic tree based on the deduced full-length amino acid sequence alignment of the published HMW-GS genes showed that the 1By16 was clustered with Glu-1B-2, and that the 1Bx13 was clustered with Glu-1B-1 alleles.