大鼠干扰素诱导蛋白-10真核表达质粒的构建及其在NIH 3T3细胞中的表达
Construction of an eukaryotic expression plasmid of IP10 and its expression in NIH 3T3 cells
作者：祁志荣;李明远;张林;蒋忠华;代吕霞;李虹;
Author：
收稿日期： 年卷（期）页码：2006,(03):-215-218
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：干扰素诱导蛋白10(IP-10);NIH3T3细胞;真核表达;自身免疫性疾病;免疫荧光
Key words：
基金项目：国家自然科学基金(30470613);;
教育部博士点专项基金(20040610053)
中文摘要
目的构建大鼠干扰素诱导蛋白-10(IP-10)基因的真核表达质粒,并研究其在NIH 3T3细胞中的表达情况。方法通过PCR获得实验所需的IP-10基因片段,通过酶切IP-10基因片段、真核表达质粒载体pEGFP-c1和连接反应,构建IP-10的真核表达质粒pEGFP-c1-IP-10,重组质粒经限制性内切酶、PCR及DNA序列测定证实构建成功后,用PolyFect脂质体转染NIH 3T3细胞,然后用免疫荧光技术检测pEGFP-c1-IP-10在NIH 3T3细胞中的表达。结果酶切分析、PCR及DNA序列测定证实,IP-10基因片段被成功克隆入真核表达质粒载体pEGFP-c1,免疫荧光技术检测证实,该基因能在NIH 3T3细胞中表达。结论证实IP-10基因可在NIH 3T3细胞中表达,为将其作为DNA疫苗治疗自身免疫性疾病的研究奠定了基础。
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