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以“宁杞1号”枸杞为试材,通过RT-PCR及RACE技术进行枸杞酸性转化酶基因的克隆及组织表达分析,并运用MEGA 5.0软件对植物转化酶基因进行了系统树分析。结果表明:扩增获得2 193bp的枸杞酸性转化酶基因,命名为LbSAI(GenBank:KC776575),该基因推导氨基酸序列与马铃薯、番茄、梨等酸性转化酶基因氨基酸序列同源性为68%~100%;LbSAI编码的蛋白质属于液泡酸性转化酶;Real-time PCR表达分析显示,LbSAI基因在枸杞花中表达量最高,在根中表达水平较低。
The Lycium barbarum L. gene was cloned and analyzed by RT-PCR and RACE. The phytase gene was analyzed by phylogenetic tree using MEGA 5.0 software. The results showed that the 2 193 bp gene of Lycium barbarum acid invertase was obtained by amplification and named as LbSAI (GenBank: KC776575). The amino acid sequence of this gene was 68% -100 with the nucleotide sequence of acid invertase from potato, tomato and pear %. The protein encoded by LbSAI belonged to vacuolar acid invertase. Real-time PCR analysis showed that the expression level of LbSAI gene was highest in Lycium barbarum and lower in roots.