精神分裂症易感基因DISC1重组质粒pmirGLO-DISC13′UTR的构建和分析

Construction and analysis of recombinant plasmid pmirGLO-DISC13′UTR of schizophrenia susceptibility gene DISC1

目的:构建精神分裂症断裂基因1(DISC1)重组质粒pmirGLO-DISC13′UTR,分析和鉴定该重组质粒。方法:通过生物信息学分析精神分裂症病人差异表达miRNA-181b-5p与精神分裂症易感基因DISC1的结合位点,选取包含结合位点在内的上下游200 bp长度序列DISC13′UTR(1.26 kb-WT)人工合成目的基因,将合成的目的基因导入H343 pmirGLO空载体构建pmirGLO-DISC13′UTR,对构建的重组质粒进行分析和鉴定。结果:测序结果显示,DISC13′UTR(1.26 kb-WT)基因成功插于H343 pmirGLO质粒,pmirGLO-DISC13′UTR序列正确,与目标序列比较覆盖度为84%,相似度为100%。结论:成功构建了精神分裂症易感基因DISC1重组质粒pmirGLO-DISC13′UTR,为miRNA-181b-5p调控精神分裂症易感基因DISC1奠定了基础,可用于后续研究。

Objective:To construct the recombinant plasmid of the disrupted in schizophrenia 1(DISC1)pmirGLO-DISC13′UTR,then analyze and identify the recombinant plasmid.Methods:The binding sites of differentially expressed miRNA-181b-5p in schizophrenia and schizophrenia susceptibility gene DISC1 were analyzed by bioinformatics.The upstream and downstream 200 bp length sequence of DISC13′UTR(1.26 kb-WT)including the binding sites was selected and synthesized.The synthesized target gene was inserted into H343pmirGLO plasmid to construct pmirGLO-DISC13′UTR,which was analyzed and identified.Results:The sequencing results showed that the DISC13′UTR(1.26 kb-WT)gene was successfully inserted into the H343pmirGLO plasmid.The pmirGLO-DISC13′UTR sequence was correct.Compared with the target sequence,the query cover of pmirGLO-DISC13′UTR sequence was 84%and the identity was 100%.Conclusions:The recombinant plasmid pmirGLO-DISC13′UTR is successfully constructed,which lays a foundation for the regulation of schizophrenia susceptibility gene DISC1 by miRNA-181b-5p and can be used in follow-up study.