In vivo assembly of plasmid‐expressed ribosomal protein S7 of Thermus thermophilus into Escherichia coli ribosomes and conditions of its overexpression

FEBS Letters

Karginova, Olga A.¹  Kopylov, Alexei M.¹  Spiridonova, Vera A.¹  Karginov, Andrey V.¹ 

[1] Chemistry Department, Moscow State University, 119899 Moscow, Russian Federation

关键词: Overexpression;    Ribosomal protein S7;    Assembly (in vivo);    Thermus thermophilus;    Escherichia coli;   

DOI  :  10.1016/0014-5793(95)00730-W

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Researchers still have great difficulty in isolating individual ribosomal proteins from the ribosome in quantities high enough for structural research. To this end, when studying protein S7, we created an E. coli overproducer of the recombinant protein S7 of Thermus thermophilus. The vector for expression was pQE-32 having a strong promoter of E. coli phage T5 and six triplets of His at the 5′-end. This N-terminal six His tag of the fusion protein is responsible for binding to Ni-NTA-resin and allows purifying the protein in one step. The yield of the recombinant protein was 20% and more of the total cellular proteins. In addition we have shown that the recombinant thermophilic protein is incorporated in vivo into the ribosome of E. coli despite the fact that these proteins (thermophilic and mesophilic) have a rather low homology, only 52%. This fact provides a base for the system to study functions of individual proteins.

【 授权许可】

Unknown