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精神分裂症断裂基因1(DSC1)是目前较明确的精神分裂症易感基因之一,其首次在一个具有染色体平衡易位的苏格兰精神疾病高发家系中被发现,后续研究[1]显示,DISC1与精神分裂症存在明显相关关系。精神分裂症的遗传模式复杂,难以用单纯的遗传因素来解释,目前认为可能是遗传因素和环境因素两者相互作用共同导致了精神分裂症的发生,表观遗传学解释了在不改变基因型的情况下环境因素是如何致病的。微小RNA(miRNA)作为表观遗传学的一员,可以靶向结合目标mRNA负性调控众多基因。精神分裂症关联的miRNA中,miRNA-181b表达和调控的异常与精神分裂症的发生和发展的密切关联已有报道[2-4]。miRNA-181b可能通过抑制DISC1基因及其蛋白的表达,影响突触和神经元的发育,在精神分裂症的发病过程中起重要作用。本研究拟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DISC1 3′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-DISC1 3′UTR,对构建的重组质粒进行分析和鉴定,以期为miRNA-181b-5p调控精神分裂症易感基因DISC1的机制研究提供基础。
精神分裂症易感基因DISC1重组质粒pmirGLO-DISC1 3'UTR的构建和分析
Construction and analysis of recombinant plasmid pmirGLO-DISC1 3'UTR of schizophrenia susceptibility gene DISC1
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摘要:
目的构建精神分裂症断裂基因1(DISC1)重组质粒pmirGLO-DISC1 3'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DISC1 3'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-DISC1 3'UTR,对构建的重组质粒进行分析和鉴定。 结果测序结果显示,DISC1 3'UTR(1.26 kb-WT)基因成功插于H343 pmirGLO质粒,pmirGLO-DISC1 3'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-DISC1 3'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。 -
关键词:
- 精神分裂症 /
- 精神分裂症断裂基因1 /
- 微小RNA181b-5p /
- 重组质粒
Abstract:ObjectiveTo construct the recombinant plasmid of the disrupted in schizophrenia 1(DISC1) pmirGLO-DISC1 3'UTR, then analyze and identify the recombinant plasmid. MethodsThe 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 DISC1 3'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-DISC1 3'UTR, which was analyzed and identified. ResultsThe sequencing results showed that the DISC1 3'UTR (1.26 kb-WT) gene was successfully inserted into the H343pmirGLO plasmid.The pmirGLO-DISC1 3'UTR sequence was correct.Compared with the target sequence, the query cover of pmirGLO-DISC1 3'UTR sequence was 84% and the identity was 100%. ConclusionsThe recombinant plasmid pmirGLO-DISC1 3'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. -
Key words:
- schizophrenia /
- disrupted in schizophrenia 1 /
- miRNA-181b-5p /
- recombinant plasmid
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