aFGF植物表达载体构建及转化紫花苜蓿的初步研究

草业学报 ›› 2011, Vol. 20 ›› Issue (4): 180-186.

aFGF植物表达载体构建及转化紫花苜蓿的初步研究

蒋世翠¹,王义¹,孙春玉¹,王艳芳^1,2,李校堃^1,2,张美萍^1*

1.吉林农业大学生命科学学院,吉林长春 130118;
2.国家教育部生物反应器工程中心,吉林长春 130118

收稿日期:2010-05-30 出版日期:2011-04-25 发布日期:2011-08-20
作者简介:蒋世翠(1982-),女,吉林汪清人,在读博士。E-mail:yuzhu8481744@163.com
基金资助:
十一五“863” 计划生物反应器重大专项(No.2007AA100503),吉林省科技发展重点计划(No.20070922)和高等学校科技创新工程重大项目培育资金(No.70S018)资助。

A rudimentary study of the acid fibroblast growth factor’s plant expression vector construction and transformation into Medicago sativa

JIANG Shi-cui¹, WANG Yi¹, SUN Chun-yu¹, WANG Yan-fang^1,2, LI Xiao-kun^1,2, ZHANG Mei-ping¹

1.College of Life Science, Jilin Agricultural University, Changchun 130118, China;
2.Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Changchun 130118, China

Received:2010-05-30 Online:2011-04-25 Published:2011-08-20

摘要/Abstract

摘要： 主要将aFGF基因按照植物偏爱的密码子进行基因优化,同时加上信号肽、6-组氨酸标签及凝血酶切割因子,合成全基因,再将基因插入到改造的TΩ4AB质粒中,将aFGF基因和质粒上的增强子、启动子、Ω序列及ployA加尾序列双酶切,命名为TΩ4AB-aF,将TΩ4AB-aF插入到pBI121载体中命名为pBI121-TΩ4AB-aF。利用三亲融合法将pBI121-TΩ4AB-aF转入到农杆菌菌株LBA4404中,转化苜蓿。转基因苗用卡那霉素进行筛选;并对抗性苗进行PCR、RT-PCR、Western Blot检测。结果表明,aFGF在苜蓿中得到表达。为苜蓿作为植物生物反应器奠定理论基础。

Abstract: After optimization of the aFGF gene for the chosen plant, the codon, the signal peptide, six-histone tag and zymoplasm cutting factor were added to the modified TΩAB plasmid, which was named TΩAB-aF after the recombination of the aFGF gene and enhancer sequence, promoter, Ω-sequence and ployA tailing sequence double digests. The binary expression vector pBI121-TΩAB-aF was constructed using TΩAB-aF and pBI121. By triparental mating the vector pBI121-TΩAB-aF was transformed into LBA4404. Medicago sativa was transformed using Agrobacterium tumefaciens, followed by selective screening and PCR, RT-PCR and Western Blot analysis. The aFGF was expressed in M. sativa.

中图分类号:

蒋世翠,王义,孙春玉,王艳芳,李校堃,张美萍. aFGF植物表达载体构建及转化紫花苜蓿的初步研究[J]. 草业学报, 2011, 20(4): 180-186.

JIANG Shi-cui, WANG Yi, SUN Chun-yu, WANG Yan-fang, LI Xiao-kun, ZHANG Mei-ping. A rudimentary study of the acid fibroblast growth factor’s plant expression vector construction and transformation into Medicago sativa[J]. Acta Prataculturae Sinica, 2011, 20(4): 180-186.

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