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Development and characterization of polyclonal antibodies against the linker region of the telomere-binding protein TRF2
Ilicheva, Nadya V; Travina, Aleksandra O; Voronin, Alexey P; Podgornaya, Olga I.
Afiliação
  • Ilicheva, Nadya V; Institute of Cytology RAS. Saint Petersburg. RU
  • Travina, Aleksandra O; Saint Petersburg State Institute of Technology. Saint Petersburg. RU
  • Voronin, Alexey P; Institute of Cytology RAS. Saint Petersburg. RU
  • Podgornaya, Olga I; Institute of Cytology RAS. Saint Petersburg. RU
Electron. j. biotechnol ; 32: 1-5, Mar. 2018. ilus
Article em En | LILACS | ID: biblio-1022489
Biblioteca responsável: CL1.1
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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Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Proteína 2 de Ligação a Repetições Teloméricas / Anticorpos Limite: Animals Idioma: En Revista: Electron. j. biotechnol Assunto da revista: BIOTECNOLOGIA Ano de publicação: 2018 Tipo de documento: Article / Project document País de afiliação: Federação Russa País de publicação: Chile

Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Proteína 2 de Ligação a Repetições Teloméricas / Anticorpos Limite: Animals Idioma: En Revista: Electron. j. biotechnol Assunto da revista: BIOTECNOLOGIA Ano de publicação: 2018 Tipo de documento: Article / Project document País de afiliação: Federação Russa País de publicação: Chile