Isolation and cloning of full-length cDNA of FatB genes from Camellia oleifera and its sequence analysis

Using the maturescent seeds of 'Xianglin No.1' as test materical,and adopting the degenerate PCR and RACE,the full-length cDNA of FatB gene from Camellia oleifera was obtained.Alignment of RACE fragments'sequencing results suggested that FatB genes in Camellia oleifera is not a single gene but a multigene family.The positions of initiation codons and stop codons of RACE sequences were predicted by means of bioinformatics method,and based on these information,sense primer was designed to land on the upstream of initiation codon,and antisense primer was designed to land on the downstream of stop codon.Lots of clones were acquired by RT-PCR amplification,and then by random sequencing several clones together with bioinformatics analysis,five gene members were isolated.They were named as co-fatb1,co-fatb2,co-fatb3,co-fatb4,co-fatb5 and their open reading frames were 1 272 bp,1 311 bp,1 305 bp,1 305 bp,1 263 bp in length.After submitting these sequences to GenBank,the GenBank accession numbers were assigned as FJ899670,FJ899671,FJ899672,FJ899673 and FJ899674 accordingly.Finally,some special amino acid residues of CoFatBs were marked and analyzed.