甘蔗果糖-1,6-二磷酸酯酶的cDNA片段克隆及序列分析  

Cloning and Sequence Analysis of Fructose-1,6-bisphosphatase cDNA from Sugarcane Stalk

作　　者：叶冰莹[1,2] 黄金存[1,2] 邵武华[1] 陈由强[1,2] 陈如凯[2] 

机构地区：[1]福建师范大学生命科学学院,福州350108 [2]农业部甘蔗生理生态与遗传改良重点开放实验室,福州350002

出　　处：《中国糖料》2009年第3期1-4,共4页Sugar Crops of China

基　　金：国家"948"项目(2006-G37);国家公益性行业(农业)科研专项经费项目(nyhyzx07-019;2007-2010)

摘　　要：植物生长发育所需要的光合产物大部分以蔗糖的形式供应和运输,其中果糖-1,6-二磷酸酯酶(fructose-1,6-bisphosphatase,FBPase)水解果糖-1,6-二磷酸(FBP)形成F6P和无机磷。根据GenBank登录的甘蔗细胞质型FBPase基因(GenBank登录号x89006)的序列设计引物,从甘蔗叶片cDNA文库和茎中扩增出FBPase基因的cDNA片段sfbp1和sfbp2,并构建到pMD18-T载体进行序列测定和分析。结果表明,sfbp1和sfbp2长均是1180bp,包含完整的开放读码框,编码332个氨基酸。sfbp1核苷酸序列及其推演的氨基酸序列与甘蔗FBPase基因进行序列比对,相似性均是99%。采用DNAMAN比对分析sfbp1和sfbp2的cDNA序列及其推演的氨基酸序列,结果表明,相似性分别是99.93%和99.7%。采用ExPASy软件预测推演的sfbp1的分子量大小为36.074kD,理论等电点为5.83。Sucrose is the primary photosynthesis product in plant growth to supply and transportation, and FBPase is one of the necessary key enzymes for sucrose to enter each metabolic pathways. PCR primers were designed based on the sugarcane cytoplasm fructose-1,6-bisphosphatase gene registered in GenBank （x89006）. The eDNA fragments sfbp 1 and sfbp2 of sugarcane fructose-1,6-bisphosphatase gene were cloned from sugarcane （Saccharum sp. cuhivar FN95 -1702 ）leaf eDNA library and stem. The pMD18-T-sfbpl and pMD18-T-sfbp2 vectors were eonstrueted and sequenced. The result showed that the length of sfbp1 and sfbp2 were 1180 base pairs, including open reading frame （ORF） of 996 base pairs, which contained a deduced amino acid sequence of 332 residues. The sj＇bpl ORF nucleotide sequences and deduced amino acid comparison with x89006 registered in GenBank by NCBI Blast showed an identity of 99% and 99%. The ORF nucleotide sequences and their deduced amino acid comparison between slop 1 and sfbp2 by DNAMAN blast, showed an identity of 99.93% and 99.7%. Analysis by ExPASy showed that molecular weight and theoretic isoelectrie point of the protein was 36.074 kD and 5.83, respectively.

关 键 词：甘蔗 果糖-1 6-二磷酸酯酶(FBPase) CDNA克隆 

分 类 号：S566.1[农业科学—作物学]