CAJ | 학술논문

从两份西藏青稞材料中分离克隆出4个B组醇溶蛋白基因(BH1-BH4),DNA测序结果表明:它们均包含完整的开放阅读框.推断的氨基酸序列与先前报道的大麦B组醇溶蛋白具有相同的蛋白质基本结构.系统分析表明:它们推断的氨基酸序列与栽培大麦中的B组醇溶蛋白具有较高的相关性,与野生大麦和山羊草属的醇溶谷蛋白相似性较低.并且,在4个基因BH1～BH4中,BH1与先前报道的B组醇溶蛋白基因有较低的序列相似性,因此我们对BH1基因进行了原核表达,含该基因的表达载体在大肠杆菌中表达出相对分子质量为28.15 kDa并以包涵体形式存在的蛋白,进一步对其在青稞谷粒品质改良中的潜在价值进行了探讨.
종량빈서장청과재료중분리극륭출4개B조순용단백기인(BH1-BH4),DNA측서결과표명:타문균포함완정적개방열독광.추단적안기산서렬여선전보도적대맥B조순용단백구유상동적단백질기본결구.계통분석표명:타문추단적안기산서렬여재배대맥중적B조순용단백구유교고적상관성,여야생대맥화산양초속적순용곡단백상사성교저.병차,재4개기인BH1～BH4중,BH1여선전보도적B조순용단백기인유교저적서렬상사성,인차아문대BH1기인진행료원핵표체,함해기인적표체재체재대장간균중표체출상대분자질량위28.15 kDa병이포함체형식존재적단백,진일보대기재청과곡립품질개량중적잠재개치진행료탐토.
Four B-hordein genes, designated BH1-BH4, were cloned using PCR amplification from two hull-less barley cultivars,ZQ7239 and ZQ148, collected from Tibet. The results of sequencing indicated that BH1-BH4 contained complete open reading frames (ORFs). Comparison of their predicted polypeptide sequences with the published sequences suggested that they all share the same basic protein structure. Phylogenetic analysis indicated that the deduced amino-acid sequences of BH1-BH4 genes were more closely related to B-hordeins from cultivated barley (Hordeum vulgare L.) than to any other prolamins from wild barley and Aegilops tauschii. Comparison of the coding regions of BH1-BH4 genes showed that BH1 had a lower sequence identity to other previously published B-hordeins than the other three B-hordeins obtained in this study. BH1 was then cloned in a bacterial expression vector based on bacteriophage T7 RNA polymerase. The resulting plasmid produced a 28.15 kDa protein in Escherichia coli. The potential value of B-hordein genes in grain quality improvement of hull-less barley has been discussed.