高级检索+

腾冲嗜热厌氧杆菌Cmr3的生物信息学分析及原核表达

Expression and Bioinformatics Analysis of Cmr3 in Thermoanaerobacter tengcongensis

  • 摘要: 本研究旨在阐明腾冲嗜热厌氧杆菌(Thermoanaerobacter tengcongensis)Ⅲ-B型CRISPR-Cas系统Cmr蛋白在嗜热机制中的作用,利用PCR方法扩增出腾冲嗜热厌氧杆菌的cmr3基因,构建原核表达载体,将其转化至大肠杆菌(Escherichia coli)中,通过诱导表达出Cmr3蛋白;并利用生物信息学软件分析比较cmr3基因在腾冲嗜热厌氧杆菌和两种常温菌中编码氨基酸的基本理化性质和蛋白三级结构。结果显示,腾冲嗜热厌氧杆菌Cmr3蛋白在大肠杆菌中表达,分子质量为43.4 kDa;生物信息学分析发现,腾冲嗜热厌氧杆菌cmr3基因序列长1 134 bp,共编码377个氨基酸,Cmr3蛋白具有亲水性,且与常温菌相比更具有热稳定性,蛋白三级结构更为紧凑。本研究可以为了解嗜热菌CRISPR-Cas系统在嗜热过程中的作用提供科学依据。

     

    Abstract: The aim of this study was to illuminate the role of the Cmr3 of Ⅲ-B CRISPR-Cas system of Thermoanaerobacter tengcongensis in thermal adaptation. The cmr3 was cloned by PCR, and constructed to the prokaryotic expression vector, which was transformed into Escherichia coil. The Cmr3 protein was expressed after induced.Applying bioinformatic software to analyzing comparable basic physiochemical properties and tertiary structure of cmr3 gene encoded amino acids in Thermoanaerobacter tengcongensis and two mesophiles. The results showed that:Cmr3 of T. tengcongensis was expressed in E. coli. The molecular mass is 43.4 kDa; bioinformatics analysis showed that the length of cmr3 gene was 1 134 bp, encoding 337 amino acids. The Cmr3 protein is hydrophilic. In comparison, T. tengcongensis has higher thermos ability than mesophiles, and the tertiary structure was more compact than mesophiles. The results of this study provide scientific basis for the study of the process of thermal adaptation in the CRISPR-Cas system of thermophiles.

     

/

返回文章
返回