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, China.
Yi Chuan Xue Bao. 2006 Oct;33(10):937-47. doi: 10.1016/S0379-4172(06)60128-5.
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.
利用聚合酶链式反应（PCR）扩增技术，从采自西藏的两个裸大麦品种ZQ7239和ZQ148中克隆出4个B-醇溶蛋白基因，分别命名为BH1 - BH4。测序结果表明，BH1 - BH4含有完整的开放阅读框（ORF）。将其预测的多肽序列与已发表序列进行比较，结果显示它们都具有相同的基本蛋白质结构。系统发育分析表明，BH1 - BH4基因推导的氨基酸序列与栽培大麦（Hordeum vulgare L.）的B-醇溶蛋白的亲缘关系，比与野生大麦和节节麦的任何其他醇溶蛋白都更近。对BH1 - BH4基因编码区的比较表明，与本研究中获得的其他三个B-醇溶蛋白相比，BH1与其他先前发表的B-醇溶蛋白的序列同一性较低。随后，将BH1克隆到基于噬菌体T7 RNA聚合酶的细菌表达载体中。所得质粒在大肠杆菌中产生了一种28.15 kDa的蛋白质。文中还讨论了B-醇溶蛋白基因在改善裸大麦籽粒品质方面的潜在价值。
