中文摘要：

【目的】克隆华山新麦草（Psathyrostachys huashanica）的α-醇溶蛋白基因,并对其进行生物信息学分析,构建该基因的原核表达载体,在大肠杆菌中诱导表达融合蛋白。【方法】采用同源克隆法从华山新麦草基因组DNA中分离克隆出α-醇溶蛋白基因并进行序列分析,将克隆的华山新麦草α-醇溶蛋白基因Gli-Ns-5克隆到表达载体pET-28a（＋）上,获得重组质粒pET28a-Gli-Ns转化大肠杆菌BL21（DE3）并诱导表达。【结果】从华山新麦草基因组DNA中克隆了4个α-醇溶蛋白基因：Gli-Ns-2（FJ713595）、Gli-Ns-3（GQ139525）、Gli-Ns-4（GQ139526）和Gli-Ns-5（GQ139527）。序列分析表明,4条序列具有α-醇溶蛋白的典型结构特征,含有8个或9个半胱氨酸残基,序列FJ713595为假基因。利用所构建的大肠杆菌表达载体,经IPTG诱导,华山新麦草α-醇溶蛋白基因Gli-Ns-5（GQ139527）可在原核系统中特异性表达。Western-blot证实融合蛋白可成功表达。【结论】克隆了4个华山新麦草的α-醇溶蛋白基因序列,基因Gli-Ns-5（GQ139527）可在原核表达系统中成功表达,为小麦品质改良提供了新的候选基因。

英文摘要：

【Objective】The present study aimed at cloning and analyzing the α-gliadin genes from Psathyrostachys huashanica and expressing it in E.coli.【Method】The α-gliadin genes were amplified from P.huashanica by AS-PCR and then the cloned gene Gli-Ns-5 was inserted into pET-28a（＋）.The recombinant plasmids pET28a-Gli-Ns were expressed in a prokaryotic expression system after its transformation into BL21（DE3） pLysS host strain.【Result】Four new α-glidain genes,Gli-Ns-2（FJ713595）,Gli-Ns-3（GQ139525）,Gli-Ns-4（GQ139526） and Gli-Ns-5（GQ139527）,were isolated and characterized from the genomic DNA of Psathyrostachys huashanica.Molecular structure analysis revealed that these four genes had the typical structure of α-gliadin and contained 8 or 9 cysteine residues,respectively.Two internal stop codons were identified in coding region of FJ713595,indicating that it was a pseudogene.The results of SDS-PAGE and Western-blot demonstrated that the fusion protein could express normally in the prokaryotic expression system.【Conclusion】The four α-gliadin genes of Psathyrostachys huashanica were cloned and the gene Gli-Ns-5（GQ139527） was successfully expressed in E.coli.This study could provide new candidate genes for wheat quality improvement.