偃松PpSS和PpSE基因过表达载体构建  

Construction of Overexpression Vectors of PpSS and PpSE Genes from Pinus pumila

作　　者：王思瑶 于宏影 刘洋[1] 梁雪 南方 WANG Siyao;YU Hongying;LIU Yang;LIANG Xue;NAN Fang(Harbin Research Institute of Forestry Machinery,National Forestry and Grassland Administration,Harbin 150086,Heilongjiang;Research Center of Cold Temperate Forestry,CAF,Harbin 150086,Heilongjiang;Forestry Workstation of Mudanjiang Forestry and Grassland Administration,Mudanjiang 157006,Heilongjiang)

机构地区：[1]国家林业和草原局哈尔滨林业机械研究所,黑龙江哈尔滨150086 [2]中国林业科学研究院寒温带林业研究中心,黑龙江哈尔滨150086 [3]牡丹江市林草局林业工作站,黑龙江牡丹江157006

出　　处：《温带林业研究》2024年第2期68-71,共4页Journal of Temperate Forestry Research

基　　金：新兴食用油料树种种质资源调查收集(2019FY100802-06)。

摘　　要：【目的】构建偃松甾醇代谢途径关键基因SS和SE过表达载体,为促进偃松甾醇高效合成奠定基础,为药用偃松遗传改良提供重要基因资源。【方法】以偃松的cDNA为模板,将SS和SE基因克隆至pNC-Cam1304-35s载体;重组质粒通过PCR及测序鉴定正确后,三亲杂交转入根瘤农杆菌LBA4404。【结果】抗性农杆菌PCR产物长度与预期一致,阳性菌液测序结果与目标基因序列相同。【结论】成功构建SS和SE过表达载体,重组载体的获得为进一步挖掘其在偃松甾醇代谢中的调控机制研究奠定了基础。【Objective】In order to provide technical support and theoretical basis for the genetic transformation of Pinus pumila sterols metabolic engineering,the overexpression vectors of the key genes SS and SE in the sterol metabolic pathway of Pinus pumila were constructed.【Method】The sequences of SS and SE were amplified by polymerase chain reaction(PCR)using the cDNA template of Pinus pumila.The PCR products were cloned into pNC-Cam1304-35s,after confirmation of PCR and sequencing,recombinant plasmids were transformed to Agrobacterium tumefaciens LBA4404.【Result】The length of PCR products of resistant Agrobacterium were consistent with expectations,the sequencing results of positive bacterial solution were identical with the target gene sequences.【Conclusion】The overexpression vectors of SS and SE were successfully constructed.The acquisition of recombinant vectors will lay a foundation for further research on the regulatory mechanism in the sterol metabolism.

关 键 词：偃松 SS SE 过表达 

分 类 号：S791.24[农业科学—林木遗传育种]