指天蕉beta;-1，3-葡聚糖酶基因全长cDNA的克隆及序列分析.pdfVIP

中国农业科学 2011,44(15):3134-3141 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2011.15.008 指天蕉β-1，3-葡聚糖酶基因全长cDNA 的克隆及序列分析 翟国会，阮小蕾，吴丽婷，谭小勇，李华平 （华南农业大学资源环境学院，广州 510642 ） 摘要：【目的】克隆植物中主要的防卫基因β-1, 3-葡聚糖酶基因，为该基因功能研究和有效利用提供理论 依据。【方法】通过分析芭蕉科植物的β-1,3-葡聚糖酶基因，据其保守区域设计一对简并引物，通过RACE 法从野 生指天蕉中克隆得到一个β-1,3-葡聚糖酶基因的全长 cDNA——glu。【结果】glu 全长为 1 114 bp，其推导的氨 基酸序列与 GenBank 中其它来源的β-1,3-葡聚糖酶蛋白序列的相似性为 54%—71%。其中，与芭蕉科植物 Grand Nain 的β-1,3-葡聚糖酶蛋白序列的相似性最高（71%）。【结论】在芭蕉类作物中克隆了一个新的β-1，3-葡聚糖 酶基因,为进一步研究该基因的功能并应用于作物转基因抗病育种研究奠定了基础。 关键词：指天蕉；β-1，3-葡聚糖酶；基因克隆；序列分析 Cloning and Sequence Analysis of a Full-Length cDNA of β-1, 3-Glucanase Gene from Musa pisangawake ZHAI Guo-hui, RUAN Xiao-lei, WU Li-ting, TAN Xiao-yong, LI Hua-ping (College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642) Abstract: 【Objective 】The objective of this study is to clone β -1, 3-glucanase gene which is a group of important emzymes in plant defenses against fungal diseases, and to research their resistant functions and effective applications. 【Method 】A novel DNA fragment encoding β -1, 3-glucanase was isolated from Musa pisangawake (AA) by applying a PCR strategy in which a pair of degenerated primers deduced from the two blocks with specific conserved amino acid regions in multiple plants was used. Then the full length cDNA (glu) of the fragment was obtained by rapid amplification of cDNA ends (RACE). 【Result 】The full length of glu is 1 114 bp. Compared with other sequences of β -1, 3-glucanase in GenBank, the deduced amino acid sequences of glu shared 54% -71% in homology. 【Conclusion 】A novel β -1, 3-glucanase gene was cloned from the Musa pisangawake (AA). The result reported in this paper has laid a the foundation