Construction of plant vectors with promoter and cold-induced genes in citrus and transient expression verification

【Objective】The plant expression vectors of citrus cold-induced genes and promoters was con-structed fused with YFP reporter gene.【Methods】Ptcor8 gene and its promoter p Ptcor8 in Poncirus trifoliate,homologous gene Clcor8 and its promoter p Clcor8 in Citrus limon were cloned respectively. Then thecloning vectors containing promoters and the precursor plant expression vector p ID4 were digested withtwo kinds of restriction endonucleases,and the digested products was ligated to get intermediate vector.Next,the intermediate vectors and the cloning vectors containing the cold-induced genes were digestedwith two types of restriction endonucleases and were ligated consequently to contruct recombinant plas-mid. Finally,freeze-thawing was used to transform the vector plasmid into Agrobacterium EHA105.【Re-sults】p1D4/p Ptcor8-Ptcor8::YFP,p1D4/p Ptcor8-Clcor8::YFP and p1D4/p Clcor8-Ptcor8::YFP fusion ex-pression vectors were constructed,and further transient expression analysis showed that the fused genewas successfully expressed in C. sinensis leaves.【Conclusion】The successful construction of three fusionexpression vectors will provide a basis for transforming C. limon and analyzing the function of cold-in-duced gene Ptcor8 and its promoter p Ptcor8.