关键词: 皮层神经细胞；原代培养；β-Tubulin；Ad-XIAP；RT-PCR

Abstract:

Objective         To construct the green fluorescence protein(GFP) labeled recombinant adenovirus vector carrying the XIAP gene and transfect it into cortical nerve cells．Methods       A method of primary culture cortical neurons of the C57BL/6J mouse  was developed and an  experiment cell model of the neurons was established. Morphological changes of neuron cells were observed by a light microscope. Double immunostaining of β-Tubulin and DAPI were applied to assess the culture purity. The XIAP gene recombinant adenovirus vector was constructed by the homologous recombination.  Ad-XIAP was transfected into cortical nerve cells after testing the virus titer determined by tissue culture infectious dose 50(TCID50). Results       The target gene XIAP amplified by PCR was identified by DNA sequencing. The adenoviral vector could be examined 24 hours after transfection，and the green fluorescence intensity became greatest at about 72 hours. Conclusion         The recombinant adenovirus vector containing XIAP was  successfully constructed  and a  highly efficient virus was obtained which could efficiently transfect cortical nerve cells and this  laid a foundation for following investigations．

Key words: Cortical nerve cells; Primary culture; β-Tubulin; Ad-XIAP; RT-PCR