Cloning and Sequence Analysis of Uroporphyrinogen III Synthase Gene in Wheat

A 364 bp cDNA fragment was amplified from cDNA transcriped from the total RNA of wheat using a pair of specific primers,which were designed at the highly conserved regions according to the wheat ESTs and the rice UROS gene.Taking the 364 bp cDNA fragment as querying sequence,5 highly homologous ESTs were found at the wheat EST database by Local Blast and then a 1 210 bp contig with the full ORF of UROS gene was assembled.Another pair of specific primer were designed according to this assembled contig and a specific 1 077 bp cDNA fragment was amplified.The 1 077 bp fragment was then cloned and sequenced.The cDNA sequence was disparate among different species.However,the protein had a conserved domain Hem D,or HEM 4,though the amino acid sequence shows little homolog in differrent species.The phylogenic analysis suggests that the UROS gene is disparate in the evolutionary.