Tropic1808在PC12细胞中的表达

TROPIC1808 EXPRESSING IN PC12 CELL LINE

目的构建稳定表达tropic1808基因的PC12细胞株,并通过观察稳定株中高分子量神经丝蛋白(NF-H)的表达以初步观察该基因的作用。方法构建重组真核表达载体pcDNA-HA-tropic1808,转染PC12细胞,经G418筛选、Westernblot鉴定。采用RT-PCR,实时PCR,Westernblot和细胞组织化学方法观察tropic1808稳定株中NF-H的表达。结果Westernblot检测表明,tropic1808基因稳定表达,RT-PCR,实时PCR,Westernblot和细胞组织化学方法均观察到NF-H在tropic1808稳定株中的表达高于空载体对照。结论Tropic1808基因可以在PC12稳定表达且NF-H在tropic1808稳定株中表达上升。

Objective To obtain stable-expressing cell lines which could constantly express tropiclS08 gene and investigate the effect of tropic1808 by detecting the expression level of NF-H in PC12 cells. Methods The recombinant tropic1808 plasmid was constructed and transfected into PC12 cells. PC12 cells stable-expressing tropic1808 were established by C418 selection and confirmed by Western blot.The expression of NF-H in the PC12 stable-expressing tropic1808 was observed by RT-PCR,real time PCR,Western blot analysis and cytohistochemistry methods. Results Western blot confirmed that trnpic1808 gene could express stably in PC12 cells.The data of RT-PCR, real time PCR, Western blot and cytohistochemistry all demonstrated that the expression of NF-H was increased in PC12 cells stable-expressing tropic1808. Conclusion PC12 cell lines stable - expressing tropic1808 were obtained and the expression of NF-H up-regulated in the PC12 cells stable-expressing tropic1808 gene.