Proteins-partners of PH domain of BCR-ABL protein: creation of DNA constructs to uncover molecular characteristics of CML development

  • S. V. Antonenko Institute of Molecular Biology and Genetics of Natl. Acad. Sci. of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnogo str., 150
  • I. V. Kravchuk Institute of Molecular Biology and Genetics of Natl. Acad. Sci. of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnogo str., 150
  • D. S. Gurianov Institute of Molecular Biology and Genetics of Natl. Acad. Sci. of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnogo str., 150
  • G. D. Telegeev Institute of Molecular Biology and Genetics of Natl. Acad. Sci. of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnogo str., 150

Abstract

Aim. Impact of domains of Bcr in oncogenic effect associated with Bcr-Abl remains unclear. Investigation of protein-protein interactions can be one of the effective ways to reveal those molecular events that alter normal cellular processes and cause malignant transformation. Previous research showed that USP1, Cortactin and Hsp27 may interact with PH domain. To confirm interactions and to study their biological consequences, genetic constructs for expression and microscopy should be created. Methods. Various standard molecular cloning techniques and expression in E. coli strain Rosetta. Results. Several DNA constructs have been created (pBluescriptSKII(+)+USP1, pFastFT-N1-CTTN, pMediumFT-N1-CTTN, pSlowFT-N1- CTTN and pЕТ42а-hsp27). Effective bacterial expression of Hsp27 has been performed. Conclusions. All DNA constructs can be effective instruments to study biological role of interactions between PH domain of Bcr and USP1, Cortactin, Hsp27.
Keywords: PH domain, Bcr-Abl, USP1, Cortactin, Hsp27.

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