Recombinant adenovirus Pax6 nerve cell cycle.doc

 PAGE \* MERGEFORMAT 7 Recombinant adenovirus Pax6 nerve cell cycle Of: Hu Kai width, Limei Yu, Teng Sasa, Bai Jie [Abstract] Objective To determine whether Pax6 regulates cell cycle directly. Methods Pax6 recombinant adenovirus transfected SH-SY5Y neural cells to Pax6 overexpression of cell cycle detected by flow cytometry. Results neural cell line SH-SY5Y overexpressing Pax6 did not affect the cell cycle G0/G1, G2 / M, S phase parameters, the statistics and the control group after no statistical significance. Conclusion transfected with recombinant adenovirus does not affect Pax6 SH-SY5Y cell cycle. [Keywords:] Pax6; adenovirus; cell cycle Pax6 (Paired box) is the embryonic brain expression of a transcription factor, Pax6 mutations cause human congenital aniridia patients, while in Pax6 mutant mice will result in a special small eye phenotype [1]. Literatures In the early embryonic (E12), Pax6 mutations can reduce the neural precursor cell cycle, while in the late embryonic (E15) Pax6 mutations of neural precursor makes significant growth in S-phase cell cycle variable length [2-3]. In order to clarify Pax6 molecular mechanisms regulating cell cycle, we carried out this research, the purpose is to identify whether Pax6 directly regulates nerve cell cycle. 1 Materials and methods 1.1 Construction of the virus Extraction of fetal whole brain RNA, reverse transcription, do PCR, products were Hind III / Sal I endonuclease restriction enzyme digestion, purified for use. While double digestion pDC316-eGFP plasmid, purified using T4 ligase and the above PCR product connection, transformation, pick clones, shaking bacteria, PCR positive clones were screened and sequenced proved to be the correct sequence of the Pax6 expression plasmid. this pDC316-eGFP-Pax6 and pBHGlox (delta) E1, 3cre plasmids were cotransfected with Lipofectamin2000 293A cells, 24h can be seen in the fluorescence microscope adenovirus vector transfected cells the green fluoresc