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西藏青稞4个B组醇溶蛋白基因的克隆和特征(英文)

Cloning and Characterization of Four B-hordein Genes from Tibetan Hull-less Barley (Hordeum vulgare subsp vulgare)

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【作者】 韩兆雪钱刚潘志芬邓光兵吴芳唐亚伟强小林余懋群

【Author】 HAN Zhao-Xue, QIAN Gang, PAN Zhi-Fen, DENG Guang-Bing, WU Fang, TANG Ya-Wei, QIANG Xiao-Lin, YU Mao-Qun Chengdu Institute of Biology, the Chinese Academy of Sciences, Chengdu 610041, China; Graduate School of the Chinese Academy of Sciences, Beijing 100039, China; Department of Biology, ZunYi Medical College, Zunyi 563003. China; Agricultural Institute, Tibetan Academy of Agricultural and Animal Science, Lhasa 850002, China

【机构】 中国科学院成都生物研究所西藏自治区农牧科学院农业研究所中国科学院成都生物研究所 成都 610041 中国科学院研究生院北京 100039成都 610041 遵义医学院生物教研室遵义 563003成都 610041拉萨 850002

【摘要】 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。

【Abstract】 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 Ae-gilops 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.

【基金】 This work was supported by the Knowledge Innovation Project (No. KSCX2-SW-304) the Western Light Project of the Chinese Academy of Sciences, and the Collaboration Project between Chinese Academy of Sciences and Tibet.
  • 【文献出处】 遗传学报 ,Acta Genetica Sinica , 编辑部邮箱 ,2006年10期
  • 【分类号】S512.3
  • 【被引频次】3
  • 【下载频次】168
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