作物学报2011，Vol.37Issue(1)：79-86,8.DOI:10.3724/SP.J.1006.2011.00079
小麦品种陕253γ-醇溶蛋白基因的克隆、原核表达与功能鉴定
Cloning, Prokaryotic Expression, and Functional Testing of a γ-Gliadin Gene from Wheat Cultivar Shaan 253
摘要
Abstract
The γ-gliadin gene (GenBank accession No.GQ857626) was isolated from a wheat (Triticum aestivum) cultivar with a hight quality Shaan 253, using inductuion a set of genome specific primers.The deduced amino acid sequence of GQ857626 exhibited an additional cysteine residue in the region Ⅱ of mature γ-gliadin subunits compared to the previous report.A prokaryotic expression vector was constructed to express GQ857626 in the host bacterium Escherichia coli Rosetta-gami B (DE3).The SDS-PAGE and Western blot analysis confirmed that the fusion protein was expressed by induction of 0.1 mmol L-1 isopropyl-β-D-thiogalactoside (IPTG), and its actual molecular weight was about 50 kD nearly equal to the predicted.After separation and purification using HisTrap HP affinity chromatography, the function of fusion protein was tested by means of micro 4 g farinograph.When the subunit of GQ857626 was integrated into the control flour through oxidation-reduction reaction, many important farinograph parameters changed drastically, such as development time, dough stabilizing time, degree of softening.Although the gluten strength of the control flour could be improved in some degree, the subunit of GQ857626 showed a negative effect on the rheological properties of wheat flour.关键词
普通小麦/γ-醇溶蛋白/原核表达/粉质参数引用本文复制引用
王明霞,高翔,陈其皎,董剑,赵万春,李艳亮,李敏..小麦品种陕253γ-醇溶蛋白基因的克隆、原核表达与功能鉴定[J].作物学报,2011,37(1):79-86,8.基金项目
本研究由国家自然科学基金青年科学基金项目(30900896),陕西省"13115"科技创新工程重大项目(2007ZDKG-01),现代农业产业技术体系建设专项(NYCYTX-001)和西北农林科技大学基本科研业务费青年项目(QN2009007)资助. （30900896）
