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
王思瑶 1于宏影 1刘洋 2梁雪 3南方2
作者信息
- 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;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
- 2. 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
- 3. 牡丹江市林草局林业工作站,黑龙江牡丹江 157006
- 折叠
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.
Abstract
[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Key words
Pinus pumila/SS/SE/overexpression引用本文复制引用
出版年
2024
NETL
NSTL
万方数据