AIM: To clone rat transcription factor Nkx2.2 and construct its eukaryotic expression vector, express it in C6 glioma cells. METHODS: RT-PCR was used to amplify fragment including Nkx2.2 gene code region from rat brain RNA, the PCR product was inserted into pMD20-T vector, then amplified Nkx2.2 coding region from pMD20-T-Nkx2.2 and inserted it into eukaryotic expression vector pcDNA3.1-myc-his( - ). After being confirmed by sequencing, the pcDNA3.1-Nkx2.2 was transfected into C6 glioma cells. The expression of Nkx2. 2 in C6 cells was detected by indirect immunofluorescent staining. RESULTS: Sequence analysis conformed that Nkx2. 2 was correctly inserted into pcDNA3.1-myc-his( - ) vector, C6 cells transfected with Nkx2.2 could efficiently express Nkx2.2 in cell nuclear. CONCLUSION: The cloning of rat Nkx2.2 and the construction of its eukaryotic expression vector were successful, Nkx2.2 was efficiently expressed in C6 cell nuclear. This study would lay the foundation for further research on the function of Nkx2.2 and its role in nervous system.
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万方数据
维普
