›› 2010, Vol. 49 ›› Issue (增刊01): 94-98.

• 论文 • 上一篇    


管玉霞1,罗 泓1,王锡平2   

  1. (1. 福州大学生物科学与工程学院, 福建 福州 350108;2. 上海强耀生物科技有限公司,上海 200000)
  • 收稿日期:2010-02-24 修回日期:1900-01-01

Solidphase Synthesis of Polypeptide TATPTD and Its Interaction with DNA

GUAN Yuxia1, LUO Hong1, WANG Xiping2   

  1. (1.College of Biological Science and Technology, Fuzhou University, Fuzhou,Fujian 350002, China;2.China Peptides Co. Lt,Shanghai 20000, China)
  • Received:2010-02-24 Revised:1900-01-01

摘要: 通过N-芴甲氧羰基(N-fluorenylmethoxycarbonyl,Fmoc) 作为 α氨基的保护基, 以逐个延伸的固相合成法合成TAT-PTD多肽,运用高效液相色谱和质谱鉴定合成多肽的纯度和相对分子质量。通过紫外吸收光谱和琼脂糖凝胶电泳阻滞实验研究合成的TAT-PTD多肽与质粒DNA相互作用,实验表明TAT-PTD多肽与质粒DNA可以通过静电作用相互吸引、靠近并结合,于是为下一步小单孢菌基因工程的细胞转导提供了依据。

关键词: 小单孢菌, TATPTD多肽, 质粒DNA, 琼脂糖凝胶阻滞电泳, 紫外吸收光谱

Abstract: Polypeptide TAT-PTD was synthesized by solidphase method with florenl methyoxycarbonyl amino acids elongated one by one. We identified the purity of synthetic peptides and molecular weight with the help of highperformance liquid chromatography (HPLC) and mass spectrometry.We used the UV absorption spectrometry and electrophoresis mobility shift assay to study the interaction of polypeptide TAT-PTD with plasmid DNA .It showed that they combined with each other by electrostatic interaction. So it could be used for the cell transduction in the field of genetic engineering, especially in Micromonospora.

Key words: Micromonospora, polypeptide TAT-PTD, plasmid DNA, electrophoresis mobility shift assay, UV absorption spectrometry