摘要
目的构建GATA-3特异性短发夹状RNA(shRNA)逆转录病毒表达载体。方法设计、合成两对GATA-3编码基因的反向重复序列,中间插入9个碱基的短发夹,经退火形成互补双链,再克隆至逆转录病毒载体RNA i-Ready pSIREN-RetroQ。结果构建的重组逆转录病毒载体可以被限制性内切酶BamH I,EcoR I切开,电泳可显示相应条带,经测序其序列与设计的一致。结论成功构建GATA-3特异性shRNA逆转录病毒载体RetroQ/GATA-3/shRNA,为进一步进行肿瘤细胞GATA-3基因沉默的研究奠定了基础。
Objective To construct and identify recombinant retroviral vectors of shRNA specific for GATA-3. Methods Two 19bp reverse repeated motifs of GATA-3 targeting sequence with 9bp spacer were synthesized respectively and inserted into retroviral vector RNAi-Ready pSIREN-Retro-Q. Results The recombinant retroviral vectors digesied by BamH I, EcoR I showed the corresponding bands, and their sequences were the same to the antici- pation. Conclusion The recombinant retroviral vectors of pSIREN-Retro-Q/GATA-3/siRNA-1, pSIREN-Retro-Q/ GATA-3/siRNA-2 specific for GATA-3 are constructed successfully,which contribute to the study of GATA-3 gene silencing.
出处
《中国临床保健杂志》
CAS
2006年第3期254-256,共3页
Chinese Journal of Clinical Healthcare
基金
安徽省自然科学基金资助项目(20050430701)
安徽省教育厅自然科学研究计划项目(2005KJ256)
关键词
转录因子
肿瘤细胞
培养的
RNA干扰
遗传载体
Transcription factors
Tumor cells, cultured
RNA interference
Genetic vectors
