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小鼠白细胞介素2基因克隆及其逆转录病毒载体的构建

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摘要 从小鼠脾脏淋巴细胞中抽提总RNA,应用RT-PCR技术扩增出mIL-2基因,按常规方法构建T载体,然后克隆、酶切鉴定、测序分析;通过双酶切将该基因片段接入逆转录病毒载体pGEZ-Term中,脂质体法共转染包装细胞293T;用含病毒颗粒的293T细胞上清液感染L929细胞,加入Zeocin进行筛选,挑选出能稳定表达抗性的L929细胞株,并通过RT-PCR技术检测到转基因L929细胞株中mIL-2基因的转录,表明成功构建了含mIL-2基因的重组逆转录病毒载体并筛选出整合有mIL-2基因的细胞株。
出处 《江苏农业科学》 CSCD 北大核心 2008年第1期48-51,共4页 Jiangsu Agricultural Sciences
基金 国家科技型企业技术创新基金(编号:04C26223200112)
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参考文献6

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