中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
CHINESE JOURNAL OF IMMUNOLOGY
2015年
5期
650-654
,共5页
刘文明%杨晶晶%胡如意%邱兴烽%石春燕%齐忠权%刘忠臣%庄国洪
劉文明%楊晶晶%鬍如意%邱興烽%石春燕%齊忠權%劉忠臣%莊國洪
류문명%양정정%호여의%구흥봉%석춘연%제충권%류충신%장국홍
TNFAIP8%RNA干扰%重组质粒%筛选
TNFAIP8%RNA榦擾%重組質粒%篩選
TNFAIP8%RNA간우%중조질립%사선
TNFAIP8%RNA interference%Recombinant plasmid%Screen
目的：构建并筛选出干扰效率最佳的TNFAIP8-shRNA-pSIREN-RetroQ干扰质粒。方法：通过生物软件选择3个TNFAIP8基因干扰位点，构建干扰质粒并测序验证，将干扰质粒及对照质粒分别转染至 A549细胞，通过RT-PCR、Western blot检测干扰效率。结果：经RT-PCR和Western－Blot证实TNFAIP8-shRNA-pSIREN-RetroQ干扰质粒能有效干扰并抑制细胞内TNFAIP8基因的表达，通过流式检测发现降低TNVAIP8表达可以提高细胞对aDR5 ScFv诱导凋亡的敏感性。结论：成功构建和设计了对TNFAIP8基因具有显著干扰效率的干扰质粒，为进一步研究TNFAIP8基因的功能奠定了基础。
目的：構建併篩選齣榦擾效率最佳的TNFAIP8-shRNA-pSIREN-RetroQ榦擾質粒。方法：通過生物軟件選擇3箇TNFAIP8基因榦擾位點，構建榦擾質粒併測序驗證，將榦擾質粒及對照質粒分彆轉染至 A549細胞，通過RT-PCR、Western blot檢測榦擾效率。結果：經RT-PCR和Western－Blot證實TNFAIP8-shRNA-pSIREN-RetroQ榦擾質粒能有效榦擾併抑製細胞內TNFAIP8基因的錶達，通過流式檢測髮現降低TNVAIP8錶達可以提高細胞對aDR5 ScFv誘導凋亡的敏感性。結論：成功構建和設計瞭對TNFAIP8基因具有顯著榦擾效率的榦擾質粒，為進一步研究TNFAIP8基因的功能奠定瞭基礎。
목적：구건병사선출간우효솔최가적TNFAIP8-shRNA-pSIREN-RetroQ간우질립。방법：통과생물연건선택3개TNFAIP8기인간우위점，구건간우질립병측서험증，장간우질립급대조질립분별전염지 A549세포，통과RT-PCR、Western blot검측간우효솔。결과：경RT-PCR화Western－Blot증실TNFAIP8-shRNA-pSIREN-RetroQ간우질립능유효간우병억제세포내TNFAIP8기인적표체，통과류식검측발현강저TNVAIP8표체가이제고세포대aDR5 ScFv유도조망적민감성。결론：성공구건화설계료대TNFAIP8기인구유현저간우효솔적간우질립，위진일보연구TNFAIP8기인적공능전정료기출。
Objective:To construct and screen the high efficiency interference plasmid of TFAIP8-shRNA-pSIREN-RetroQ.Methods:Selected and synthesized three Target Sequence of TNFAIP8 shRNA1,TNFAIP8 shRNA2,TNFAIP8 shRNA3,and construct the TNFAIP8 interference plasmid.Transfection TNFAIP8-shRNA-pSIREN-RetroQ interference plasmid to A549 cells.Filter out the highest interference efficiency plasmid by detecting the mRNA and protein levels using RT-PCR and Western blot methods.Results:We successfully design and built three TNFAIP8-shRNA-pSIREN-RetroQ interference plasmids,and screen out the highest efficiency interference plasmid.Conclusion: Three interference plasmids targeting the TNFAIP8 gene have been constructed successfully and provide a useful tool for studying the function of TNFAIP8.
