甜瓜果实蔗糖磷酸合成酶基因cDNA片段的克隆及表达分析  

Cloning and Expression Analysis of a Sucrose Phosphate Synthase cDNA Fragment in Melon Fruit

作　　者：于喜艳[1] 樊继德[1] 何启伟[2] 

机构地区：[1]山东农业大学园艺科学与工程学院,山东泰安271018 [2]山东省农业科学院蔬菜所,济南250100

出　　处：《园艺学报》2007年第1期205-208,共4页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30471191);山东省良种产业化课题;山东农业大学博士基金项目

摘　　要：根据在GenBank中登录的番茄、马铃薯等蔗糖磷酸合成酶基因的保守序列设计引物,采用RT-PCR方法从甜瓜花后25d的果实总RNA中扩增出目标cDNA片段,克隆到pMD18-T载体中。序列分析表明,该片段与番茄蔗糖磷酸合成酶氨基酸序列同源性为98.9%,GenBank中登记号为DQ355797。通过Northernblot检测其在甜瓜果实不同发育时期的表达变化,结果表明该基因在甜瓜果实花后25d开始表达,随着果实的成熟,表达量升高。PCR primers were designed based on the conserved domain of some sucrose phosphate synthase genes in GenBank. The target cDNA fragment was amplified from the total RNA isolated from melon fruit of 25 days after pollination with RT-PCR and then was cloned into pMD18-T vector. The sequence showed that the amino acid sequence encoded by this fragment showed a 98.9% similarity to that of the corresponding region of sucrose phosphate synthase gene in tomato. The accession number of this gene in GenBank was DQ355797. Northern blot was performed to analyze the expression pattern of sucrose phosphate synthase in different development stages of melon fruit. The result showed that sucrose phosphate synthase began to express in fruit 25 days after pollination and the expression level of this gene increased with melon fruit mature.

关 键 词：甜瓜 蔗糖磷酸合成酶基因 克隆 基因表达 

分 类 号：S652[农业科学—果树学]