Construction of Gateway Plant Expression Vectors with xylA Gene as Selection Marker

Objective:To construct plant expression vector with xylose isomerase gene xylA as the selection marker used in Gateway technology system.Methods:First,xylA gene was cloned from Escherichia coli,and was constructed into pCAMBIA1301 by substituting for hpt gene.Second,the fragment including P35S,T35S,attR1,at-tR2 and CmR-ccdB of the Gateway Binary Vector(pH7WG2D)was cut by XbaⅠand HindⅢand was recombinanted into the expression vector pCAMBIA1301-xylA.Third,the donor vector with HY5 gene[cloned from‘Tsuda’Turnip(Brassica campestris L.ssp.rapa)after BP reaction]was mixed with pCAMBIA1301-xylA-GW vector by LR reaction.Then,the recombinant plasmid was transformed into Agrobacterium tumefaciens LBA4404 by electroporation.Results:The plant expression vector pCAMBIA1301-xylA-HY5 for Gateway technology system was successfully constructed by identification.Conclusion:The results showed that it is convenient to construct non-antibiotic marker plant expression vector by using xylA as selective marker and Gateway cloning technology.