大豆7S球蛋白(α+β)-亚基缺失型种质的α-与β-亚基基因的测序与分析.pdfVIP

中国农业科学 2009,42(2):419-424 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2009.02.005 大豆7S球蛋白(α+β)-亚基缺失型种质的 α-与 β-亚基基因的测序与分析 刘珊珊，孟凡立，刁桂珠，王志坤，葛玉君，郑天慧，姜自芹，曾 蕊，李文滨 （东北农业大学农学院大豆研究所/ 国家教育部大豆生物学重点实验室，哈尔滨 150030 ） 摘要：【目的】明确大豆7S球蛋白（α+β）-亚基双缺失特性是否是由于α-与β-亚基基因的缺失或变异引 起的。【方法】聚丙烯酰胺凝胶电泳(SDS)分析和α-与β-亚基基因的PCR扩增、克隆、测序和序列比较。【结 果】该种质α-亚基基因特异性引物的PCR 扩增结果与对照基本一致，β-亚基基因的则与对照不同。α-亚基基因 扩增片段与对照的同源性较高，为 90.9%, 二者的区别主要存在于扩增片段的 5′-端；β-亚基基因间的同源性较 低，仅为 53.1%。另外，在该材料β-亚基基因扩增片段的两端，检测到一对短的反向重复序列。【结论】该种质 α-与β-亚基同时缺失的特性不是由α-与β-亚基基因的缺失引起的；α-与β-亚基基因的扩增序列与对照存在 一定的差异，基因序列间的差异是否为导致α-与β-亚基缺失的直接原因，尚需进一步研究确认。 关键词：大豆；7S球蛋白；α-与β-亚基；稀有亚基组合；DNA测序 Sequence Analysis of α- and β-Subunit Genes in a Novel Soybean Germplasm Characterized by Absence of α- and β-Subunit of 7S Globulin LIU Shan-shan, MENG Fan-li, DIAO Gui-zhu, WANG Zhi-kun, GE Yu-jun, ZHENG Tian-hui, JIANG Zi-qin, ZENG Rui, LI Wen-bin (Soybean Research Institute of Northeast Agricultural University Ministry of Education Key Laborabory of Soybean Biology, Harbin 150030) Abstract: 【Objective 】 The objective of this study was to elucidate whether the α- and β- null character was result from the absence or variance of α- and β-subunit genes. 【Method 】 SDStechnique, PCR amplification, cloning and sequencing methods were used. 【Result 】 With specific primers for α- subunit, the PCR band pattern between (α+β)-null type and normal type was identical, but the PCR band pattern of β-subunit gene was unique. For PCR amplified DNA fragment of the α-subunit gene, a high degree of nucleotide homology (90.9%) exists between the (α+β)-null type and normal typ