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油菜Bnpgip2-1基因的克隆表达与生物信息学分析
Cloning, prokaryotic expression and bioinformatics analysis of polygalacturonase-inhibiting protein gene (Bnpgip2-1) from rape
据GenBank及相关文献提供的序列设计引物，从油菜基因组DNA中扩增出Bnpgip2-1基因完整开放阅读框。该基因全长1 068bp，与已发表的Bnpgip2基因序列（登录号：AF529694）有98%的相似性。RT-PCR分析表明，该基因（已申请序列号为KJ820998）具一个72bp的内含子（544-615），编码区为996bp，含4个限制性内切酶酶切位点（Ava Ⅰ，185；Hind Ⅲ，536；EcoR Ⅰ，929；ApaL Ⅰ，972）。原核表达该基因，表达产物能显著抑制油菜菌核病菌PG活性，抑制率达52.96%。生物信息学分析表明，表达产物BnPGIP2-1有331个氨基酸，理论分子量为36.99kDa，pI为8.35，具强疏水性，主要存在于细胞壁。信号肽切点位于第22和23位氨基酸之间（SFS-KNL），N端和C端各存在5个和3个半胱氨酸残基，可形成3个二硫键；二级结构以α-螺旋（31.12%）、β折叠（16.01%）和无规则线圈（52.87%）为结构元件，具典型的LRR结构；三级结构为10个LRR按右手螺旋规则形成的一个开放裂隙区，可能负责着其与病菌PG的互作。该基因的表达受病原菌侵染的强烈诱导，但对水杨酸（SA）处理不敏感，茉莉酸（JA）处理反而使其下调表达。
多聚半乳糖醛酸酶抑制蛋白 / 原核表达 / 生物信息学分析 / 核盘菌 {{custom_keyword}} /
Polygalacturonase-inhibiting protein / prokaryotic expression / bioinformatics analysis / Sclerotinia sclerotiorum {{custom_keyword}} /
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江苏省自然科学基金（BK2010305）
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