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Vol 32 (2018)
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Development and characterization of polyclonal antibodies against the linker region of the telomere-binding protein TRF2 | Ilicheva | Electronic Journal of Biotechnology
doi:10.1016/j.ejbt.2017.12.001
Electronic Journal of Biotechnology, Vol 32 (2018)

Development and characterization of polyclonal antibodies against the linker region of the telomere-binding protein TRF2

Nadya Ilicheva, Aleksandra Travina, Alexey Voronin, Olga Podgornaya



Abstract

Background: TRF2 (telomeric repeat binding factor 2) is an essential component of the telomere-binding protein complex shelterin. TRF2 induces the formation of a special structure of telomeric DNA and counteracts activation of DNA damage-response pathways telomeres. TRF2 has a poorly characterized linker region (udTRF2) between its homodimerization and DNA-binding domains. Some lines of evidence have shown that this region could be involved in TRF2 interaction with nuclear lamina.

Results: In this study, the fragment of the TERF2 gene encoding udTRF2 domain of telomere-binding protein TRF2 was produced by PCR and cloned into the pET32a vector. The resulting plasmid pET32a-udTRF2 was used for the expression of the recombinant udTRF2 in E. coli RosettaBlue (DE3). The protein was isolated and purified using ammonium sulfate precipitation followed by ion-exchange chromatography. The purified recombinant protein udTRF2 was injected into guinea pigs to generate polyclonal antibodies. The ability of anti-udTRF2 antibodies to bind endogenous TRF2 in human skin fibroblasts was tested by western blotting and immunofluorescent staining.

Conclusions: In this study, the recombinant protein udTRF2 and antibodies to it were generated. Both protein and antibodies will provide a useful tool for investigation of the functions of the udTRF2 domain and its role in the interaction between TRF2 and nuclear lamina.




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ISSN:  0717-3458

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