转基因红花中角质细胞生长因子KGF-1的表达

江莺; 刘秀明; 马吉胜; 李巍; 朱海林; 杜美丽; 李海燕; 李校堃

doi:10.3724/SP.J.1259.2011.00311

植物学报 >

2011 , Vol. 46 >Issue 3: 311 - 318

DOI: https://doi.org/10.3724/SP.J.1259.2011.00311

研究报告

转基因红花中角质细胞生长因子KGF-1的表达

江莺 ,
刘秀明 ,
马吉胜 ,
李巍 ,
朱海林 ,
杜美丽 ,
李海燕 ,
李校堃

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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, 长春 130118;
²吉林农业大学中药材学院, 长春 130118
³吉林农业大学生命科学学院, 长春 130118

收稿日期: 2010-12-01

修回日期: 2011-01-24

网络出版日期: 2011-05-26

基金资助

国家高技术研究发展计划(863计划);吉林省科技发展计划重点项目;高等学校科技创新工程重大项目培育资金项目

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Expression of Keratinocyte Growth Factor-1 (KGF-1) in Transgenic Safflower

JIANG Ying ,
LIU Xiu-Meng ,
MA Ji-Qing ,
LI Wei ,
ZHU Hai-Lin ,
DU Mei-Li ,
LI Hai-Yan ,
LI Jiao-Kun

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¹Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development,Jilin Agricultural University, Changchun 130118, China;

²College of Chinese Medicinal, Jilin Agricultural University,Changchun 130118, China;

³College of Life Sciences, Jilin Agricultural University, Changchun 130118, China

Received date: 2010-12-01

Revised date: 2011-01-24

Online published: 2011-05-26

Supported by

National High Technology Research and Development Program of China (863 Program);the Science and Technology Development Projects of Jinlin Province;Important Projects in the Scientific Innovation of Colleges and Universities in China

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摘要

通过构建重组表达质粒载体p139035S-KGF1和根癌农杆菌介导在红花(Carthamus tinctorius)中表达角质细胞生长因子(KGF-1)。从侵染到诱导生根共需要14周, 转化率达0.1%。红花子叶在潮霉素筛选培养基上培养4–5周后便可获得丛生芽, 再生芽移入含潮霉素的伸长生根培养基, 培养4–8周可诱导生根。通过PCR、Southern blot、RT-PCR及Western blot检测证明目的基因KGF-1已经整合到红花细胞的染色体中, 实现了KGF-1外源蛋白在红花中的成功表达, 为开发KGF-1蛋白新的生产途径奠定了基础。

本文引用格式

江莺 , 刘秀明 , 马吉胜 , 李巍 , 朱海林 , 杜美丽 , 李海燕 , 李校堃 . 转基因红花中角质细胞生长因子KGF-1的表达[J]. 植物学报, 2011 , 46(3) : 311 -318 . DOI: 10.3724/SP.J.1259.2011.00311

Abstract

The recombinant plasmid vector p139035S-KGF1 was constructed and transfected into safflower by Agrobacterium tumefaciens transfection. The period of infection to rooting was 14 weeks, and transformation efficiency was about 0.1%. Adventitious shoots were generated after cotyledons were cultured in selecting media containing hygromycin for 4–5 weeks. Rooted safflower was obtained in 4–8 weeks. KGF-1 was transformed into safflower chromosomes and expressed successfully, as determined by PCR, Southern blot, RT-PCR and Western blot analysis, which established the basis for the production of KGF-1.

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