国际肿瘤学杂志
國際腫瘤學雜誌
국제종류학잡지
JOURNAL OF INTERNATIONAL ONCOLOGY
2008年
12期
947-950
,共4页
沈绍华%刘爱华%亓春花%叶欣%顾龙君
瀋紹華%劉愛華%亓春花%葉訢%顧龍君
침소화%류애화%기춘화%협흔%고룡군
RNA干扰%磷酸酪氨酸%信号传导
RNA榦擾%燐痠酪氨痠%信號傳導
RNA간우%린산락안산%신호전도
RNA interference%Phosphotyrosine%Signal transduction
目的 构建CBKL基因RNA干扰(RNAi)慢病毒载体.方法 将筛选确定的CRKL基因RNAi有效靶序列克隆到pGCL-GFP载体[含U6启动子和绿色荧光蛋白(GFP)]连接产生LV-shCRKL慢病毒载体.通过酶切、测序验证后,用LV-shCRKL、pHelper 1.0和pHelper2.0质粒共转染包装细胞293T细胞,包装产生慢病毒,以293T细胞GFP蛋白的表达水平测定病毒滴度.结果 载体片段插入正确.共转染包装细胞293T能产生高浓度的重组慢病毒LV-shCRKL.结论 成功构建了人CRKL基因RNAi病毒载体.
目的 構建CBKL基因RNA榦擾(RNAi)慢病毒載體.方法 將篩選確定的CRKL基因RNAi有效靶序列剋隆到pGCL-GFP載體[含U6啟動子和綠色熒光蛋白(GFP)]連接產生LV-shCRKL慢病毒載體.通過酶切、測序驗證後,用LV-shCRKL、pHelper 1.0和pHelper2.0質粒共轉染包裝細胞293T細胞,包裝產生慢病毒,以293T細胞GFP蛋白的錶達水平測定病毒滴度.結果 載體片段插入正確.共轉染包裝細胞293T能產生高濃度的重組慢病毒LV-shCRKL.結論 成功構建瞭人CRKL基因RNAi病毒載體.
목적 구건CBKL기인RNA간우(RNAi)만병독재체.방법 장사선학정적CRKL기인RNAi유효파서렬극륭도pGCL-GFP재체[함U6계동자화록색형광단백(GFP)]련접산생LV-shCRKL만병독재체.통과매절、측서험증후,용LV-shCRKL、pHelper 1.0화pHelper2.0질립공전염포장세포293T세포,포장산생만병독,이293T세포GFP단백적표체수평측정병독적도.결과 재체편단삽입정학.공전염포장세포293T능산생고농도적중조만병독LV-shCRKL.결론 성공구건료인CRKL기인RNAi병독재체.
Objective To construct a lentiviral vector carrying CRKL gene RNA interfering( RNAi ).Methods The CRKL RNAi was selected and subcloned into the lentiviral vector,pGCL-GFP(including U6 promotor and green fluorescent protein),generating the lentiviral vector LV-shCRKL.The corrected CRKL was confirmed by endoenzyme digestion ,sequencing.Recombinant lentiviruses were produced by 293T cells following the eo-transfection of LV-shCRKL,with the packaging plasmids pHelper1.0 and pHelper2.0.The virus titer was detected by GFP expressions in 293T cells.Results Plasmid LV-shCRKL carried the correct sequence.The recombinant lentiviruse LV-shCRKL could be produced by co-transfection of LV-shCRKL to 293T cells.Conclusion The recombinant lentiviruse vector LV-shCRKL is constructed successful.
