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利用简并PCR技术和RACE技术克隆得到了一条洋葱的苯丙氨酸解氨酶(PAL)基因全长cDNA序列。该cDNA序列全长2349 bp,编码长685个氨基酸残基的多肽序列,命名为AcPAL2。Blast分析表明该序列与虎眼万年青Galtonia saundersiae、野蕉Musa balbisiana的相似性均较高。Real-time PCR表达及花青素含量分析表明,红皮洋葱该基因表达量最大,而黄皮和白皮洋葱表达量极低;在红皮洋葱中该基因在膨大初期大量表达,并迅速降低至一定程度后趋于相对平稳表达,且与花青素的积累过程相一致。
The full-length cDNA sequence of phenylalanine ammonia-lyase (PAL) gene was cloned using degenerate PCR and RACE techniques. The cDNA sequence of 2349 bp in length, encoding a polypeptide sequence of 685 amino acid residues, named AcPAL2. Blast analysis showed that the sequence was highly similar to Ornithogalum Galtonia saundersiae and Musa balbisiana. The results of Real-time PCR and anthocyanin analysis showed that the expression of this gene was the highest in onion and the expression of yellow and white onions was extremely low. In onion, the gene was highly expressed in early stage of expansion and rapidly decreased To a certain extent tends to be relatively stable expression, and consistent with the accumulation of anthocyanins.