目的 利用RNA干扰(RNA interfering,RNAi)技术,构建HnRNPB1表达载体,并进行鉴定。方法 采用H1启动子,分别把被7bP序列间隔的21bp长短的HnRNPB1靶序列的反向重复序列,置于PsiRNA-hH1neoG2质粒中,分别构建HnRNPB1短发夹环RNA(shRNA),产生重组质粒PSiRNA—hHl neoG2 HnRNPB1。结果 将合成的DNA序列退火后克隆到载体上,并作测序分析,经测序鉴定确定为所需序列。结论 针对HnRNPB1基因的特异性ShRNA真核表达载体PSiR—NA—hH1 neoG2 HnRNPB1的成功构建,为进一步研究其基因功能打下基础。
Objective To clone the RNA expression vector affecting gene HnRNPB1 translation by RNA interfering and analyze the nucleic acid sequence of the erpressien vector for further searching new gene therapy method of tumor.Methods A plasmid psiRNA - hHineoG2 containing human H1 SnRNA promoter was lingated to a 21bp reverse repeated motif of HnRNPB1 target sequence with 7bp spacer, respectively. Enzyme digestion and DNA sequencing were used to examine whether the expressien vector HnRNPB1 were correct by sequence analysis. Results The expression vector was cloned and the aim sequence was obtained. Conclusion Successful cloning of the recombinant helps to search new therapy method of tumour.