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Construction and expressions of eukaryotic expression vectors of human p65 and IκBα gene in MH7a cells
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 19
- DOI:
- 10.3321/j.issn:1000-2790.2006.19.001
- Key Word:
- NF-κB;p65;IκBα;类风湿性关节炎;逆转录聚合酶链反应
Abstract： 目的: 构建人p65和IκBα基因的真核表达载体并检测其在人永生化滑膜细胞MH7a中的表达. 方法: 应用RT-PCR方法从体外培养的HEK293细胞mRNA中扩增出p65和IκBα基因的编码区,克隆至pMD18T载体并测序,然后亚克隆至真核表达载体pcDNA3.1myc/His(-)A,酶切鉴定正确后以脂质体法瞬时转染MH7a细胞,Western Blot检测p65和IκBα的表达. 结果: 测序证实PCR扩增得到的p65和IκBα基因编码区序列正确;脂质体法转染MH7a细胞后检测到预期目的蛋白的表达. 结论: 成功构建了p65和IκBα的真核表达载体并使其在人永生化滑膜细胞中表达.
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