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