枸杞酸性转化酶基因的克隆及组织表达分析

Cloning and Tissues Expression Analysis of Soluble Acid Invertase Gene from Lycium barbarumL.

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MEGA5．0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2193bp的枸杞酸性转化酶基因，命名为LbSAJ（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％；LbSAJ编码的蛋白质属于液泡酸性转化酶；Real—timePCR表达分析显示，L6SA，基因在枸杞花中表达量最高，在根中表达水平较低。

Using Lycium barbarum L. ＇Ningqi No. 1＇ as material, cloning and tissue expression analysis of SAI genes of Lycium barbarurn L. was studied,and using MEGA 5.0 software to SAI genes for phylogenetic tree was analyzed. The results showed that by RT-PCR and RACE methods a SAI gene named as Lb SAI （GenBank number： KC776575） was identified from Lycium barbarum L. the full length cDNA of Lb SAI was 2 193 bp;the amino acid sequences of Lb SAI were 68%-100% identical to the sequences of potato, tomato, pear etc. The putative protein encoded by Lb ,SAI was belonged to cell wall invertase. Real-time PCR analysis showed that the Lb SAI gene was the highest expressed in flower, the expression level reached the lowest level in root.