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目的研究脑脓肿形成和发展过程中所涉及的新分子。方法对应用mRNA荧光差异显示技术(DD-PCR)获得的脑脓肿早期差异表达cDNA片段G2-3进行电子克隆,根据电子克隆的结果设计引物进行逆转录-多聚酶链式反应(RT-PCR)、克隆及测序,验证电子克隆的准确性,并利用生物信息学技术对电子克隆产物进行功能预测。结果G2-3经电子克隆获得长度为1157bp的新基因RBAG2-3,Genbank登陆号为CN382815。结构功能分析显示RBAG2-3编码73个氨基酸残基的蛋白,存在多个功能作用位点,并具有PSI和H-NS结构功能域。结论经电子克隆获得新基因RBAG2-3,其编码蛋白具有多个功能作用位点,并具有能对神经上皮组织基因表达起作用的结构功能域,可能在脑脓肿的发病中起重要作用。
Objective To study new molecules involved in the formation and development of brain abscess. Methods The cDNA fragment G2-3 differentially expressed in brain abscess obtained by DD-PCR was cloned electronically. The primers were designed according to the results of the electronic cloning and used for reverse transcriptase-polymerase chain reaction (RT-PCR) Cloning and sequencing, to verify the accuracy of electronic cloning, and the use of bioinformatics technology for functional prediction of electronic cloning products. Results The new gene RBAG2-3 with a length of 1157bp was obtained by electron microscopy from G2-3. The Genbank accession number is CN382815. Structural and functional analysis showed that RBAG2-3 encodes a protein of 73 amino acid residues with multiple functional sites and PSI and H-NS structural domains. Conclusion The new gene RBAG2-3 obtained by electron cloning has multiple functional sites and possesses structural domains which can affect the gene expression of neural epithelium and may play an important role in the pathogenesis of brain abscess.