木薯MeGWD3基因克隆及植物表达载体构建

发布时间：2018-06-12 23:21

  本文选题：木薯 + 葡聚糖水合双激酶　； 参考：《南方农业学报》2016年07期

【摘要】：【目的】克隆木薯葡聚糖水合双激酶(Me GWD3)基因并构建其植物表达载体,为开展木薯Me GWD3基因调控及功能研究提供参考。【方法】采用RT-PCR从木薯华南124(SC124)中克隆Me GWD3基因,对其进行序列分析,并将其连接至改造的p CAMBIA2300载体,构建植物表达载体p CAMBIA2300-Me GWD3。【结果】Me GWD3基因全长3522 bp,编码1173个氨基酸。经序列比对,发现Me GWD3基因序列与木薯Phytozome数据库中公布的GWD基因(cassava4.1_000497m)只存在2个碱基差异,同源性高达99.94%;与NCBI数据库中收录的木薯GWD基因(JN618460)同源性高达99.00%,与蓖麻GWD基因(XM_002518566)同源性达86.00%;成功构建植物表达载体p CAMBIA2300-Me GWD3。【结论】成功构建的植物表达载体p CAMBIA2300可用于Me GWD3基因功能及木薯淀粉代谢研究。
[Abstract]:[objective] to clone and construct the plant expression vector of cassava dextran hydrated double kinase me GWD3 gene, and to provide a reference for studying the regulation and function of cassava me GWD3 gene. [methods] Me GWD3 gene was cloned from cassava South China 124GWD3 gene by RT-PCR. It was sequenced and ligated to the modified pCAMBIA2300 vector to construct the plant expression vector pCAMBIA2300-Me GWD3. [results] the full length of me GWD3 gene was 3522 BP, encoding 1173 amino acids. By sequence alignment, it was found that there were only two bases difference between me GWD3 gene sequence and cassava4.1000497 m published in cassava Phytozome database. The homology of GWD gene of cassava was 99.00 and 86.00 with the gene of castor GWD. The plant expression vector pCAMBIA2300-MGWD3was successfully constructed. [conclusion] the plant expression vector pCAMBIA2300-MGWD30.Conclusion the plant expression vector pCAMBIA2300 can be constructed successfully by the method of construction of the plant expression vector pCAMBIA2300. [conclusion] the plant expression vector pCAMBIA2300 can be successfully constructed by constructing the plant expression vector pCAMBIA2300. [conclusion] the plant expression vector pCAMBIA2300 can be constructed successfully. It was used to study the function of me GWD3 gene and cassava starch metabolism.
【作者单位】： 中国热带农业科学院热带生物技术研究所;
【基金】：海南省自然科学基金项目(314123)
【分类号】：S533
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本文编号：2011493