水稻 OsPIN1b 基因 GFP 融合表达载体构建及转化水稻的研究

    Construction and transformation of GFP fusion expression vector of rice OsPIN1b gene

    • 摘要: 水稻生长素输出载体 OsPIN1a 可能参与调控水稻根负向光性反应。 为了进一步研究其他 OsPIN基因与水稻根负向光性的关系, 以 GenBank 数据库报道的 OsPIN1b 核苷酸序列为基础, 设计特异引物以 RT-PCR 从水稻 cDNA 中扩增得到 OsPIN1b 基因。 测序结果表明获得完整的 OsPIN1b 基因 ORF 序列; 生物信息学分析表明, OsPIN1b 基因序列全长 1 665 bp, 与已报道的序列相吻合, 其 GC 含量为 64.08%, 编码 554 个氨基酸。 进一步构建了该基因的 GFP 融合表达载体 pCAMBIA-1301-OsPIN1b∷GFP 并转化水稻中花 11, 分子检测和 GUS 染色表明外源片段已经成功整合到水稻基因组, 为研究 OsPIN1b 在水稻根负向光性中的作用奠定基础。

       

      Abstract: Earlier research results showed that rice auxin efflux protein OsPIN1a probably participated in regulating negative phototropism of rice roots. To explore the relationship between other OsPIN genes and negative phototropism of rice roots, gene-specific primers were used to amplify OsPIN1b gene by RT-PCR based on the OsPIN1b sequence in GenBank. Sequencing result showed that the full ORF was obtained(1 665 bp). Bioinformation analysis showed that the sequence was 100% identity with the reported sequence, the GC content of OsPIN1b ORF was 64.08% and OsPIN1b encoded 554 amino acid residues. The OsPIN1b-green fluorescent protein (gfp)fusion expression vector pCAMBIA-1301-OsPIN1b∷GFP was constructed and transformed into Japonica rice Zhonghua11 by Agrobacterium tumefaciens. Molecular detection and GUS assay showed that the target construct was integrated into the genome of rice, which laid the foundation for further study of the function of OsPIN1b in regulating negative phototropism of rice roots.

       

    /

    返回文章
    返回