Construction,Transformation And Expression Analysis Of Fusion Expression Vectors Of AtHGO/OsHGO Promoter And GUS

The HGO gene encodes the 1,2-homogentisate dioxygenase,which functions in the Tyr degradation pathway.In previous study,we found that SSCD1 gene encodes an Arabidopsis putative FAH(famarylacetoacetate hydrolase)and the sscd1(short-day sensitive cell death1)mutant displays cell death under short-day condition,If hgo mutant and sscd1 mutant were hybridized,the phenotype was suppressed of sscd1 hgo mutant of Arabidopsis thaliana under short-day condition.The results showed that low concentration of tyrosine could promote the growth of Arabidopsis seedlings,and the high concentration of tyrosine could inhibit the growth of seedlings.Rice and Arabidopsis were model of monocotyledon and dicotyledon,and rice HGO gene has similar function with Arabidopsis HGO gene.In order to investigate the expression of HGO gene in plants,we performed the following studies:1.Cloning of AtHGO and OsHGO promoter.By PCR and sequencing,these two promotors sequences were cloned correctly and completely.2.Construction of the fusion expression vector of AtHGO/OsHGO promoter and GUS.The recombination with PC1301 vector containing the GUS gene was carried out using the cloned AtHGO and OsHGO gene promoter sequence to fuse with the GUS gene.3.The recombinant vector was transformed into wild-type Col-0,sscd1 and hgo mutants by Agrobacterium mediated and histochemical GUS assay.The results showed that the promoter activated the expression of GUS gene,and the expression level and location of the AtHGO and OsHGO in Arabidopsis wild-type Col-0,sscd1 and hgo mutants were different.