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Construction of NIH3T3 Cells Expressing PENK Gene by Retroviral Expression Vector

机译:逆转录病毒表达载体表达钢基因NIH3T3细胞的构建

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Objective Carrying the proenkephalin gene by recombinant retrovirus vector,to research if the gene can expressed stably in NIH3T3 cell line. Methods pLNCX2-pENK was transfected into PT67 packaging cell by Lipofectamine mediation and resistant clone was selected by G418.the viral particles that existed in the supernatants were collected, they were used to infect target cell, resistant clone was alao selected by G418.Finally, the proenkephalin gene expression was detected by immunoprecipitation and RT-PCR. Results The proenkephalin gene can integrate into NIH3T3 cell line and express established by radioimmunoassay and RT-PCR. Conclusion The proenkephalin gene can expressing stably, it could contribute to further research for the treatment of chronic pain.
机译:通过重组逆转录病毒载体携带Prenkephalin基因的目的,研究如果基因可以在NIH3T3细胞系中稳定地表达。方法通过G418将PLNCX2-PECK转染到PT67包装电池中。收集在上清液中存在的病毒颗粒,用于感染靶细胞,抗克隆选择通过G418选择。最后,通过免疫沉淀和RT-PCR检测前妥啡素基因表达。结果前奈基素基因可以集成到NIH3T3细胞系中,并通过放射免疫测定和RT-PCR建立的表达。结论前妥Kalin基因可以稳定表达,有助于进一步研究治疗慢性疼痛。

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