中国科学院上海有机化学研究所机构知识库
Advanced  
SIOC OpenIR  > 生命有机化学国家重点实验室  > 期刊论文
学科主题: 生命有机化学
题名: 丙糖磷酸异构酶、果糖-1,6-二磷酸醛缩酶及果糖-1,6-二磷酸酶的共表达、
其他题名: Co-expression of triosephosphate isomerase, fructose-1,6-bisphosphate aldolase and fructose-1,6-bisphosphatase in E.coli
作者: 唐功利 ; 杨春松 ; 鲍建绍 ; 王燕芳 ; 陈海宝
通讯作者: 陈海宝
刊名: 生物化学与生物物理学报
发表日期: 2001-01-01
卷: 33, 期:1, 页:131-136
收录类别: SCI
部门归属: 中国科学院上海有机化学研究所生命有机化学实验室; 中国科学院上海植物研究所光合室
摘要: 致力于建立一条控制或降低大气中CO^2浓度的途径,选择对蓝藻进行代谢工程以便改进其光合固定CO^2的效率。作为研究的初始阶段,将编码丙糖磷酸异构酶、果树1,6—二磷酸醛缩酶及果糖。L,6-二磷酸酶的3个基因构建进一个由启动子trc控制的表达质粒pTrcFAT,成功地在大肠杆菌中实现了 上述3个基因的活性共表达。活性测定结果显示:从I L培养液获得的破菌上清液每分钟可以催化:经丙酮磷酸(DHAP))转化成700μmol果糖—6—磷酸。在此基础上进一步构建了这3个基因共表达的大肠杆菌蓝藻穿梭表达质粒、也在大肠杆菌中实现了活性表达。当外源基因的操纵子与裁体质料以大于1:1的比例进行构建时,可显著提高外源基因的表达量及相应的酶活性。
英文摘要: To establish a way to control or to decrease the daily increasing concentration of atmospheric CO^2, metabolically engineering Cyanobacteria was taken for the improveent of its efficiency of photosynthetic CO^2fixation. As a preliminary stage of this study, three genes coding for three important Calvin cycle enzymes, I.e. Triosephosphate isomerase (TPI), fructose-1,6-bisphosphate aldolase (FBP aldolase), and fructose-1,6-bisphosphatase (FBPase), respectively, have been cloned into one plasmid, pTrcFAT, which is controlled by promoter trc. Successful co-transcriptional expression of these three genes resulted in high yields of these enzymes under the induction of 0.25 mmol/L IPTG. Bioassay showed that the expressed enzymes from one liter of culture could directly catalyze DHAP conversion into 700 μmol of fructose-6-phosphate (F-6-P) per one minute. Furthermore, in order to introduce the three genes co-expression system into Cyanobacteria, a shuttle pasmid be tween E. Coli and Cyanobacteria was constructed using plasmid pTrcFAT and a shuttle vector Pdc-8, forming a shuttle plasmid Pdcfat-2 containing a dimer of the three genes co-expression operator. Successful co-expression in E. Coli of Pdcfat-2 with higher full activity has been obtained. This shuttle was used to transform of Cyanobacteria Symechococcus sp. PCC 7942, and a few positive colonies were obtained.
语种: 中文
内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/25745
Appears in Collections:生命有机化学国家重点实验室_期刊论文

Files in This Item: Download All
File Name/ File Size Content Type Version Access License
2001024.pdf(719KB)----开放获取--View Download

Recommended Citation:
唐功利,杨春松,鲍建绍,等. 丙糖磷酸异构酶、果糖-1,6-二磷酸醛缩酶及果糖-1,6-二磷酸酶的共表达、[J]. 生物化学与生物物理学报,2001,33(1):131-136.
Service
Recommend this item
Sava as my favorate item
Show this item's statistics
Export Endnote File
Google Scholar
Similar articles in Google Scholar
[唐功利]'s Articles
[杨春松]'s Articles
[鲍建绍]'s Articles
CSDL cross search
Similar articles in CSDL Cross Search
[唐功利]‘s Articles
[杨春松]‘s Articles
[鲍建绍]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit
文件名: 2001024.pdf
格式: Adobe PDF
此文件暂不支持浏览
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Powered by CSpace