利用RT-PCR及RACE技术,从药用植物枸杞中克隆了1个编码蔗糖磷酸合成酶(SPS)基因的全长cDNA,命名为LbSPS(GenBank登录号KC834608)。序列分析表明:LbSPS基因长3 677 bp,开放阅读框为3 165 bp,编码1 033个氨基酸,分子量为118.457 5 kD,理论等电点6.05。系统进化分析显示,LbSPS编码的氨基酸序列与甜瓜、马铃薯、番茄等蔗糖磷酸合成酶基因编码氨基酸序列一致性为66%~98%。qRT-PCR分析显示,LbSPS基因在枸杞花中表达量最高,叶中表达水平较低。该研究为进一步了解LbSPS在枸杞生长发育、逆境胁迫等过程中的生物学功能奠定了基础。
Using RT-PCR and RACE methods,we cloned a SPS gene from a medicinally plant Lycium barbarum L.,named as LbSPS (GenBank number:KC834608).Sequence analyses showed that the full length cDNA of LbSPS was 3 677 bp and the 3 165 bp ORF encoded 1 033 amino acids with a molecular weight of 118.457 5 kD and an isoelectric point of 6.05.Phylogenetic analysis showed that the amino acid sequences of LbSPS were 66%~98% identical to the sequences of melon,tomato,potato,etc.qRT-PCR analysis showed that the LbSPS gene was highly expressed in flower.The expression level reached the lowest in leaf.Molecular cloning of LbSPS will be useful for further functional elucidation of the gene involving in the development,stress response in L.barbarum L..