葡萄VvBAP1基因的克隆及表达特性分析

Gene Cloning and Expression Analysis of VvBAP1 in Vitis vinifera

以抗性葡萄品种‘F-242’组培苗为材料,利用同源克隆法克隆了葡萄VvBAP1基因。测序结果显示,VvBAP1扩增片段大小为531 bp,可编码176个氨基酸序列。利用生物信息学分析VvBAP1基因编码的蛋白序列显示,该蛋白分子量为19.43kDa,含有保守的钙离子依赖性的C2结构域;等电点pI为9.42;不稳定系数为37.09,推测为稳定的亲水性蛋白;含有多个丝氨酸/苏氨酸磷酸化位点。实时荧光定量PCR表明,该基因在根茎叶中均有表达,其中在叶片中表达量较高;盐胁迫、低温等逆境因子及逆境相关的信号物质,如水杨酸和一氧化氮均可诱导VvBAP1的表达,其中低温对其表达量影响更为显著,推测该基因参与了葡萄抵御逆境胁迫的过程,尤其是与低温相关的过程。

Using homology cloning method, the full·Length cDNA of VvBAP1 was cloned from leaves of Fitis vinifera cultivar ＇F-242＇ tissue culture seedling. Bioinformatic analysis indicated that VvBAP1 consists of 531 nucleotides, encoding 176 amino acids with molecular weight 19.43 kDa, isoelectric point 9.42, and suggesting a stable hydrophilic protein which contains several serine/threonine phosphorylation sites with a C2-1ike domain, respectively. Real-time PCR analysis showed that the FvBAP1 was expressed in all tested tissues, with the highest expression in leaves. FvBAP1 was significantly induced by signal molecule in stress such as salicylic acid （SA） and nitric oxide （NO）. Moreover, the VvBAPI was accumulated in response to salt stress and cold stress, especially the low temperature. It suggested that the VvBAP1 was involved in various stresses in grape, in particular, the response to low temperature process.