四川农业大学学报 ›› 2011, Vol. 29 ›› Issue (02): 141-146.doi: 10.3969/j.issn.1000-2650.2011.02.001

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阿拉拉特小麦高分子量谷蛋白新基因1Gy的克隆及序列分析

徐黎黎, 钟美玉, 李伟, 魏育明   

  1. 四川农业大学小麦研究所, 四川 温江 611130
  • 收稿日期:2011-03-08 出版日期:2011-06-30 发布日期:2017-04-11
  • 通讯作者: 魏育明(E-mail:yuming71@hotmail.com) E-mail:yuming71@hotmail.com
  • 基金资助:
    四川省青年科技基金项目(2010JQ0046)

Cloning and Sequence Analysis of a Novel HMW-GS Gene 1Gy from Triticum araraticum Jakubz.

XU Li-li, ZHONG Mei-yu, LI Wei, WEI Yu-ming   

  1. Triticeae Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan, China
  • Received:2011-03-08 Online:2011-06-30 Published:2017-04-11

摘要: 利用y-型高分子量谷蛋白亚基的特异引物,对阿拉拉特小麦(PI427305)的基因组DNA进行PCR扩增,得到大小为2.2 kb的目的条带,将该条带回收纯化并克隆到pMD18-T载体中,经梯度亚克隆测序拼接,得到编码区的全序列为2 202 bp(GenBank登录号:HM131806),共编码732个氨基酸。它与1Gy7*序列的同源性高达99%,而且氨基酸序列结构与大多数y-型亚基相同,推断该基因为1Gy。1Gy的分子量比1Gy7*稍小,迁移率比1Gy7*慢。与小麦属其他基因组编码的y-型亚基相比,其在靠近C端的重复区多了一个半胱氨酸残基。利用在线PSIPRED对其二级结构预测结果显示,其重复区主要是无规则卷曲结构。这些结构都可能使得1Gy对小麦加工品质产生正面影响。

关键词: 阿拉拉特小麦, 1Gy, 分子克隆, 原核表达, 序列分析

Abstract: In this study, genomic PCR reaction of Triticum araraticum(PI427305) was carried out by using degenerate primers specific for y-type HMW-GS.A 2.2 kb DNA fragment was amplified and cloned into the pMD18-T vector.DNA sequencing was carried out by using overlapping sub-clones.The complete coding region of the gene was 2 202 bp(GenBank accession number was HM131806), and 732 amino acid residues were encoded.It was found that the DNA sequence of this gene showed the highest identity with that of 1Gy7* of Triticum timopheevii Zhuk, and the corresponding amino acid sequence had the typical structure of y-type HMW-GS.So it was concluded that this gene was 1Gy.The molecular weight of 1Gy was a little smaller than that of 1Gy7*, and the mobility of 1Gy was slower than that of 1Gy7*.Therefore, 1Gy was a novel y-type HMW-GS gene.Compared with other y-type subunits encoded by A, B and D genomes of common wheat, an extra cysteine existed in the repetitive domain near C-terminal domain of 1Gy.The repetitive domain was mainly proved to be coil by using PSIPRED.Both of them were beneficial to bread improvement.

Key words: Triticum araraticum, 1Gy, molecular cloning, prokaryotic expression, sequence analysis

中图分类号: 

  • S512.1