Construction of Arabidopsis CDR6 Over-expression Carrier and Identification of Stably Transgenic Lines

[Objective] To build up the carrier of Arabidopsis CDR6 gene over-expression and isolate corresponding transgenic lines. [Method]Total RNA was extracted from Arabidopsis seedlings and reverse transcribed as PCR template. CDS fragments of CDR6 gene were amplified through RT- PCR. Using the restriction endonuclease and T4 DNA ligase,CDS fragments were subsequently cloned into p XB094 vector,and then were transformed into Trans-T1 phage resistant competent cells. Bacterial colony PCR and DNA sequencing were performed to confirm that CDS of the Arabidopsis CDR6 gene was successfully cloned. The recombinant plasmids were transformed into Agrobacterium GV3101 cells. Wild type plants were transformed using floral-dip method and screened to obtain the desired transgenic plants. [Result] Bacterial colony PCR and DNA sequencing were performed and recombinant plasmids were confirmed. Stably transgenic lines of CDR6 over-expression were obtained through antibiotic screening,and verified through genetic methods. [Conclusion] Construction of Arabidopsis CDR6 gene over-expression and screening of transgenic plants laid the foundation for functional analysis of CDR6.