白桦BpGT14基因表达载体及RNA干扰载体的构建

Construction of BpGT14 gene expression vector and RNA interference vector in Betula platyphylla

糖基转移酶基因是生物分子糖基化中的关键基因,已有研究表明少数糖基转移酶14(Glycosyltransferase14,GT14)家族基因在植物细胞壁合成及对逆境的响应中发挥重要的生物学功能,然而GT14基因对逆境的响应机制目前仍不清楚。为了研究糖基转移酶基因的生物学功能,本实验室在克隆了白桦Bp GT14基因(JQ409354)编码区序列全长的基础上,利用Bam HⅠ单酶切的方法构建了白桦Bp GT14基因的p BI121植物表达载体,并利用一步PCR之后在同一体系中同时酶切-连接的方法,构建了Bp GT14基因RNA干扰载体,相比于传统的RNA干扰载体构建方法更为高效快捷。测序结果表明,植物表达载体及RNA干扰载体均构建成功。本研究的完成,为Bp GT14基因进一步的遗传转化及转基因植株的获得奠定了基础,同时为今后植物表达载体的构建提供了高效便捷的参考方法。

Glycosyltransferase gene is the key gene in the biomolecules glycosylation. Some studies have shown that a minority glycosyltransferase 14 （GT14） gene family play an important biological function in the synthesis of plant cell wall and response to adversity, but the stress response mechanism of GT14 gene remains unclear. In order to study the biological function of glycosyltransferase gene, based on full-length of cloned BpGT14 gene （JQ409354） by our laboratory, we have constructed the plant expression vector pBI121 through single digestion and connection and RNA interference vector through a newly one-step PCR method instead of the original one. Sequencing results showed the success of plant expression vector and RNAi vector. The success of this experiment laid a foundation further for the step of genetic transformation and transgenosis and provides an efficient and convenient method of the plant expression vectors constructed for future reference.