中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
8期
1071-1073
,共3页
树突状细胞%siRNA%CD86%免疫耐受
樹突狀細胞%siRNA%CD86%免疫耐受
수돌상세포%siRNA%CD86%면역내수
Dendritic cell%siRNA%CD86%Immune tolerance
目的 筛选构建针对大鼠树突状细胞表面共刺激分子CD86的小干扰RNA(siRNA)慢病毒载体.方法 设计针对CD86基因的4条siRNA序列及1条阴性对照序列,分别构建慢病毒载体,与包含CD86基因序列的质粒共转染293T细胞,Western blot检测CD86分子表达,确定最佳siRNA序列.结果 Western blot方法显示,在所选4条siRNA序列中以2号序列抑制效果最为明显,且成功构建了siRNA慢病毒载体.结论 成功构建了针对CD86分子的siRNA慢病毒载体,为该病毒的功能研究及体内实验奠定了基础.
目的 篩選構建針對大鼠樹突狀細胞錶麵共刺激分子CD86的小榦擾RNA(siRNA)慢病毒載體.方法 設計針對CD86基因的4條siRNA序列及1條陰性對照序列,分彆構建慢病毒載體,與包含CD86基因序列的質粒共轉染293T細胞,Western blot檢測CD86分子錶達,確定最佳siRNA序列.結果 Western blot方法顯示,在所選4條siRNA序列中以2號序列抑製效果最為明顯,且成功構建瞭siRNA慢病毒載體.結論 成功構建瞭針對CD86分子的siRNA慢病毒載體,為該病毒的功能研究及體內實驗奠定瞭基礎.
목적 사선구건침대대서수돌상세포표면공자격분자CD86적소간우RNA(siRNA)만병독재체.방법 설계침대CD86기인적4조siRNA서렬급1조음성대조서렬,분별구건만병독재체,여포함CD86기인서렬적질립공전염293T세포,Western blot검측CD86분자표체,학정최가siRNA서렬.결과 Western blot방법현시,재소선4조siRNA서렬중이2호서렬억제효과최위명현,차성공구건료siRNA만병독재체.결론 성공구건료침대CD86분자적siRNA만병독재체,위해병독적공능연구급체내실험전정료기출.
Objective To construct siRNA lentiviral vector targeting CD86 costimulatory molecule in dendritic cells or rats, and detect its knockdown rate. Methods Four siRNA sequences tarteting CD86 gene and a negative control sequence were designed to construct lentiviral vector, and co-transfected into 293T cells with the plasmid containing rat CD86 gene. Western blotting was used to detect the knockdown rate, and determine the optimal sequence. Results Western blotting showed in the chosen 4 sequences, the 2nd sequence had the highest knockdown rate. Conclusion The lentiviral vector targeting the CD86 gene in dentritic cells (DCs) was successfully constructed.
