Recombinant production of Thermus aquaticus single-strand binding protein for usage as PCR enhancer

Year 2021, Volume: 5 Issue: 1, 42 - 46, 15.04.2021

Özlem Kaplan , Rizvan İmamoğlu , İskender Şahingöz İsa Gökçe

https://doi.org/10.35860/iarej.766741

Cited By: 4

Abstract

Single-stranded DNA-binding (SSB) proteins play an important role in DNA metabolism involving DNA replication, recombination, and repair in all living beings. In molecular biology, SSB proteins are used as enhancers to increase the efficiency and specificity of PCR. Thermostable SSB protein eliminates secondary structure or dimer formation and significantly increase the effectiveness of amplification of DNA fragments. In this study, it was ensured that the SSB gene of thermophilic bacteria Thermus aquaticus (T. aquaticus) was cloned into the pET28b vector and expressed in E. coli BL21 (DE3) PLysE cells. Then, the purification of the SSB protein produced in E. coli BL21 (DE3) PLysE cells was performed. 20 mg SSB protein was obtained from 1L bacterial culture, and its purity was more than 90%. It was shown by the PCR experiment that the SSB protein produced in this study could increase the amplification efficiency.

Keywords

Polymerase Chain Reaction, Single Strand Binding Protein, Thermus aquaticus

Supporting Institution

Turkish Scientific and Technical Research Council (TUBITAK)

Project Number

TUBITAK-2209A

Thanks

We thank to TUBITAK for supporting this work.

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