Transfection and expression of the recombinant adeno-associated virus carrying human interferon-γ in human lymphocyte

ZHU Haihong, CHEN Zhi, ZHANG Mingtai, et al.
（Institute of Infectious Diseases,the First Affiliated Hospital,Medical college of Zhejiang University,Hangzhou 310003, China）

　　【Abstract】　Objective　In order to study the transfection and expression of the recombinant pWIG of Adeno-associated virus and Human interferon-γ(HuIFN-γ) in human lymphocyte.Methods　After transfecting pWIG into human lymphocyte via calcium phosphate, DNA, RNA and protein extracted from transfected cells were detected by polymerase chain reaction(PCR), reverse transcription PCR(RT-PCR) and ELISA. Results　The 504 bp bands were detected from PCR and RT-PCR products of the DNA and RNA from the cells transfected, and HuIFN-γ protein were detected to be 17.2 pg/3×10⁶ cells by ELISA.Conclusion　The recombinant pWIG could produce HuIFN-γ mRNA after the transfection via calcium phosphate, and translate corresponding protein further. This lays ground for the further research of gene therapy.
　　【Key words】Gene therapy; Interferon γ; Adeno-associated virus; Lymphocyte

　　腺相关病毒(AAV)是目前基因治疗的常用载体系统之一。因其具有稳定表达、定点整合、安全性较高等优点而受到重视。我们在以往工作的基础上^［1］，将已构建成功的腺相关病毒和人γ-干扰素(HuIFN-γ)基因的重组体pWIG转染源于人淋巴细胞的H9细胞株，在DNA、RNA、及蛋白水平探讨其表达状态。