摘要
以苦荞基因组DNA为模板,根据已构建的苦荞eDNA文库中蔗糖合酶(Sucrose synthase,SuSy)的部分eDNA序列设计引物,PCR扩增SuSy基因的核心片段,结合Genomic walking技术获得SuSy基因的完整序列。生物信息学分析表明,该基因序列全长4428bp,含有12个外显子和11个内含子,剪接位点均符合经典GU—AG法则,ORF长度为2412bp,共编码803个氨基酸。其氨基酸序列与籽粒苋(Amaranthus hypo—chondriacus)、甜菜(Beta vulgaris)、红叶藜(Chenopodium rubrumL)和大豆(Glycine max)的相似性分别为86%、86%、85%和83%。保守结构域分析表明,SuSy含有1个GT1糖基化酶功能结构域(275~760)和4个ADP结合位点,能够催化果糖-6-磷酸和尿苷5L二磷酸葡萄糖形成蔗糖-6-磷酸。
We designed the primers according to the cDNA sequence of the sucrose synthase gene (Su- Sy) obtained from cDNA library of buckwheat in laboratory. The core fragment of the sucrose synthase gene (SuSy) was amplified by PCR and a fullqength DNA sequence was cloned by Genomic Walking. Bioinformatics analysis showed that the full-length of SuSy gene of Fagopyrum tatarium was 4 428 bp including 11 introns and 12 exons. The ORF of SuSy gene was 2 412 bp,probably encoding a protein of 803 amino acids. Further analysis indicated that the deduced SuSy protein had homology of 86 %,86 %,0,85 % and 83 % with Amaranthus hyDochondriacus, Beta vulgaris, ChenoDodiurn rubrum L,Glycine rnaz. Protein conserved domain searching results showed that SuSy contained a GT1 sucrose synthase domain and four putative ADP-binding pockets which may catalyze the synthesis of sucrose 6-phosphate from fructose 6-phosphate and uridine 5P-diphosphate-glucose. This result laid the foundation for further exploration of the expression regulation of sucrose synthase in tartary buckwheat.
出处
《西北农业学报》
CAS
CSCD
北大核心
2015年第11期80-86,共7页
Acta Agriculturae Boreali-occidentalis Sinica
基金
国家自然科学基金(31171606)~~
关键词
苦荞
蔗糖合酶
基因克隆
生物信息学分析
Fagopyrum tatarium
Sucrose synthase
Gene cloning
Bioinformatics analysis