Abstract:

BACKGROUND: The basic helix-loop-helix transcription factor oligodendrocytes transcription factor 2 is a key regulator for the differentiation of oligodendrocyte lineage cells during development.
OBJECTIVE: To construct the recombinant eukaryotic expression vector of rat oligodendrocyte transcription factor 2, and to investigate its expression in eukaryocytes.
METHODS: Oligodendrocyte transcription factor 2 cDNA fragments were cloned by reverse transcription-PCR with the total RNA from neonatal rat spinal cord as the template, and subsequently cloned into pGEM-T vector. The oligodendrocyte transcription factor 2 fragment with its sequence confirmed was then cloned into pEGFP-N1 vector directionally. The right recombinant was transfected into COS-7 cells by lipofectamine 2000. The expression of oligodendrocyte transcription factor 2 in COS-7 cells was detected by reverse transcription-PCR and immunoblot analysis.
RESULTS AND CONCLUSION: Oligodendrocyte transcription factor 2 cDNA was cloned, and the correct pEGFP-N1-Olig2 cloning was verified by restriction endonuclease digestion and sequencing. The Western blot analysis indicated that Olig2-GFP fusion protein was expressed in COS-7/pEGFP-N1-Olig2 cells at 72 hours after transfection. pEGFP-N1-Olig2 was constructed successfully. Olig2-GFP fusion protein could be abundantly expressed in COS-7/pEGFP-N1-Olig2 cells.