Figure 1

Full length of Crypthecodinium cohnii AdoMet synthease. (A) The sequencing strategy for AdoMet synthetase in Crypthecodinium cohnii. Based on two partial sequences (red rectangles) obtained from the C. cohnii cDNA library, the full length AdoMetS was isolated from C. cohnii DNA by three rounds of PCR. The center region between two C. cohnii clones mentioned above was isolated by PCR using P1F (5'-ATACAGCTGATGGTCATGGTA-3') and P1R (5'-CTTGCGGCCAGT CAA-3'). The 3' region of CcAdoMetS was isolated by PCR using P2F (5'-ATCATCATCGACACGTACGGTG-3') and P2R (5'-GAGAAAGGCGGACA GGTATCC-3') with C. cohnii cDNA library constructed form C. cohnii total RNA and cloned into the pDNR-LIB vector (Clontech). The 5' region was isolated by using 5'-rapid amplification of cDNA ends (BD SMARTTM RACE cDNA Amplification Kit, ClonTech) by P3R 5'-CGGCACCGATGTCCAAGTCCTCCTTT-3'. (B) Nucleotide and deduced amino acid sequences of the Crypthecodinium cohnii AdoMet synthetase gene. Putative start codon ATG and putative stop codon TAA are shown in the block. Poly(A) tail is underlined.