关键词: 油菜, fae1, PCR, 基因克隆, 序列分析, 芥酸

Abstract: fae1 gene is a rate-limiting gene for erucic acid synthesis in Brassica. Partial sequences of the gene were cloned by polymerase chain reaction (PCR) strategy with primers F1 and R1 designed according to the DNA sequence of fae1 reported in Genbank (AF490462). Twelve cultivars with different erucic acid content from B.napus, B.oleracea and B.rapa , including 2 resynthesized B.napus lines were used as genomic DNA source materials. The PCR products were cloned into the vector pGEM-T and sequenced individually. All the 12 cultivars showed similar nucleotide sequences with a same 1007bp in length. A clustal alignment of the 12 sequences showed that fae1 gene is highly conservative among different species tested, and within the 1007bp region showed 23 sites of single nucleotide polymorphism corresponding to 11 amino acid changes at protein level. No significant difference was found between natural and resynthesized lines of B.napus. Specific SNP sites were found to distinguish fae1 genes between high and low erucic acid and between C genome and A genome.

Key words: Brassica, fae1, PCR, Gene cloning, Sequence analysis, Erucic acid