Cloning and vector construction of MADS-Box gene in Phalaenopsishybrid

ObjectiveCloning as well as sense and antisense plant expression vector of MADS-Box gene in Phalaenopsis hybrid were conducted to provide references for researching its functions. MethodThe MADS-Box gene was cloned from scape of Phalaenopsis hybrid cv. Jiuhbao Red Rose using RT-PCR and RACE. Sense and antisense plant expression vector were also constructed. ResultThe cloned MADS-Box gene from scape was named DtpsMADS1(GenBank accession No. JQ065097). The full length cDNA of DtpsMADS1 was 960 bp, containing 37 bp of a 5'-untranslated region (5'-UTR),185 bp of 3'-UTR,and 738 bp of an opening reading frame (ORF). The gene encoded 245 predicted amino acids. The bioinfor-mation analysis results showed that the gene encoding protein was a basic hydrophilic protein and contained 62.45%ofα-heli-cal domains, 8.16%of extended strand and 29.39%of random coil. Sequence comparison and phylogenetic analysis revealed that DtpsMADS1 shared 99.0%homology with ORAP13 of Phalaenopsis amabilis, 83.0%and 82.0%homology with Dendrobi-um nobile and Cymbidium orchid, respectively. DtpsMADS1 belonged to A-class subfamily of MADS family. Sense and anti-sense plant expression vector pBI121-DtpsMADS1-S and pBI121-DtpsMADS1-A were constructed when DtpsMADS1 was connected to plant expression vector pBI121. ConclusionThe DtpsMADS1 gene of Phalaenopsis hybrid was cloned suc-cessfully. The sense and antisense plant expression vector were constructed successfully and could be used for DtpsMADS1 genetic function identification and genetic improvement of Phalaenopsis hybrid.