VEGF基因油体表达系统构建及对拟南芥的遗传转化

doi:10.7506/spkx1002-6630-201009034

食品科学 ›› 2010, Vol. 31 ›› Issue (9): 144-149.doi: 10.7506/spkx1002-6630-201009034

VEGF基因油体表达系统构建及对拟南芥的遗传转化

李洪志^1,2，庞实锋³，李海燕¹，薛萍^1，刘秀明¹，李营¹，李巍¹，肖艳双^1,4，李校堃^{1 , 4 ,*}

1.吉林农业大学生物反应器与药物开发教育部工程研究中心 2.吉林农业大学生命科学学院 3.广东医学院生物教研室 4.温州医学院药学院

收稿日期:2009-08-03 修回日期:2009-12-07 出版日期:2010-05-01 发布日期:2010-12-29
通讯作者: 李洪志 E-mail:hongzhi-li2008@163.com
基金资助:
国家“863”计划项目(2007AA100503)；吉林省科技发展重点计划项目(20070922)；
高等学校科技创新工程重大项目培育资金项目(70S018)；长春市科技计划项目(06GG150)

Construction of VEGF Gene Plasmid and Transformation into Arabidopsis thaliana

LI Hong-zhi1,2，PANG Shi-feng3，LI Hai-yan1，XUE Ping1，LIU Xiu-ming1，LI Ying1，LI Wei1， XIAO Yan-shuang1,4，LI Xiao-kun1,4,*

1. Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Jilin Agricultural University,
Changchun 130118, China；2. College of Life Science, Jilin Agricultural University, Changchun 130118, China；
3. Department of Biology, Guangdong Medical College, Dongguan 523808, China；4. College of Pharmacy, Wenzhou Medical
College, Wenzhou 325027, China

Received:2009-08-03 Revised:2009-12-07 Online:2010-05-01 Published:2010-12-29

摘要/Abstract

摘要：

为研究油体系统在植物中表达外源蛋白的优势，以油菜总DNA 为模板，通过PCR 得到油菜油体(Ycoil)及启动子序列(Ycprm)。同时，将多对合成的寡核苷酸链，通过PCR 方法拼接得到血管内皮生长因子全长基因(VEGF)。以此为基础，构建Ycoil、Ycprm、VEGF 融合表达载体p1390Ycprm-Ycoil-VEGF。将重组质粒转入农杆菌GV3101，并用花浸染法对拟南芥进行浸染。在含潮霉素(20mg/L)的1/2MS 培养基上筛选转基因拟南芥种子(T0)，获得了T1 代抗性植株，通过PCR 检测、SDS-PAGE 检测和Western blotting 检测，结果表明油菜油体及血管内皮生长因子的融合基因已经转入拟南芥中，并成功得到表达，转化率约为0.1%。

关键词: 油体, 血管内皮生长因子全长基因(VEGF), 拟南芥, 载体构建, 植物生物反应器

Abstract:

In order to investigate exogenous protein expression in oil body system, the fragments of rape oleosin gene promoter (Ycprm) and rape oleosin gene (Ycoil) were obtained through PCR amplification using the designed primers and total DNA from Brassica napus L. VEGF gene was synthesized by overlap extension PCR. An oil body expression plasmid p1390Ycprm-Ycoil- VEGF was constructed by Ycprm, Ycoil and VEGF gene using the reconstruction vector, pCAMBIA1390. This recombinant plasmid was transformed into A. tumefaciens GV3101. A resistant plant, T1, was obtained in 1/2 MS medium containing hygromycin at the concentration of 20 mg/L by screening the seeds (T0) of transformed Arabidopsis plants. The results of PCR, SDS-PAGE and Western blotting showed that the fusion protein with Ycoil gene and VEGF gene were successfully expressed in Arabidopsis thaliana.

Key words: oil body, VEGF, Arabidopsis thaliana, plasmid construction, plant bioreactor

中图分类号:

Q782

李洪志1,2，庞实锋3，李海燕1，薛萍1，刘秀明1，李营1，李巍1，肖艳双1,4，李校堃1,4,*. VEGF基因油体表达系统构建及对拟南芥的遗传转化[J]. 食品科学, 2010, 31(9): 144-149.

LI Hong-zhi1,2，PANG Shi-feng3，LI Hai-yan1，XUE Ping1，LIU Xiu-ming1，LI Ying1，LI Wei1， XIAO Yan-shuang1,4，LI Xiao-kun1,4,*. Construction of VEGF Gene Plasmid and Transformation into Arabidopsis thaliana[J]. FOOD SCIENCE, 2010, 31(9): 144-149.

[1]	何胜华，邓乾春. 大豆和油菜籽油体形成的天然乳液的稳定性及胃肠道消化特性[J]. 食品科学, 2021, 42(13): 34-42.
[2]	孙禹凡，谢凤英，钟明明，齐宝坤，李杨. Oleosin蛋白和磷脂酰胆碱相互作用对重组油体乳液稳定性的影响[J]. 食品科学, 2020, 41(14): 42-49.
[3]	马涛，姚明月，刘延琳. 酿酒酵母APA1的定点突变以及与增强型绿色荧光蛋白EGFP基因的共表达[J]. 食品科学, 2015, 36(23): 200-204.
[4]	叶红红，尹明安*，朱敏，任小林. 番茄果实特异表达载体pBI121-2A11-HB-1的构建及其对农杆菌的转化[J]. 食品科学, 2014, 35(19): 174-179.
[5]	刘顺枝，孙莉丽，杨礼香，王小兰*. 八氢番茄红素脱氢酶(PDS)基因果实特异表达载体的构建[J]. 食品科学, 2010, 31(13): 196-199.

VEGF基因油体表达系统构建及对拟南芥的遗传转化

Construction of VEGF Gene Plasmid and Transformation into Arabidopsis thaliana

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 5

编辑推荐

Metrics

本文评价