抑制性底物杂交(SSH)技术研究BXSB小鼠的差异表达基因

Isolation of the differentially expressed genesin the BXSB mouse model of systemic lupus erythematosus by SSH(Suppression subtractive hybridization)technique

目的：利用抑制性底物杂交技术（Ｓｕｐｐｒｅｓｓｉｏｎ ｓｕｂｔｒａｃｔｉｖｅ ｈｙｂｒｉｄｉｚａｔｉｏｎ，ＳＳＨ），以ＢＸＳＢ 狼疮小鼠为模型，研究系统性红斑狼疮（ＳＬＥ） 发病相关基因。方法：分离ＢＸＳＢ和Ｃ５７ＢＬ６ 小鼠骨髓细胞，提取ｍＲＮＡ，反转录构建ｃＤＮＡ 文库，利用ＳＳＨ 技术研究ＢＸＳＢ小鼠差异表达基因。结果：获得了１２ 个阳性克隆，其中有３ 个克隆编码ＴＣＲ、ＭＨＣ Ⅱ类分子及逆转录病毒基因表达产物，均在ＢＸＳＢ狼疮小鼠高表达，表明其与系统性红斑狼疮发病有关；１ 个在ＢＸＳＢ狼疮小鼠高表达的基因可能为新的ｃＤＮＡ片段，已被ＧｅｎＢａｎｋ 收录，登记号码为ＡＦ０９３４５３。结论：ＴＣＲ、ＭＨＣ Ⅱ类分子及逆转录病毒基因表达产物与系统性红斑狼疮发病有关。

Objective:Toisolatethe differentiallyexpressed genesin BXSBmouse.Methods:Separated bone marrow cellsfrom the BXSB and C57 BL 6 mice,extracted the polyA+ RNA,synthesized the dscDNAby reverse transcription,andidentified the differentially expressed genes by SSHtechnique.Results:Twelve positive genes wereisolated,three ofthem have beenreportedto be relatedto SLE,including TCR, MHC Ⅱandretrovirusgene products.Inaddition,foundone novelgenesimilartothe TCRgene,submitteditto GenBank,andaccepted an ac cession number AF093453.Conclusion:TCR,MHC Ⅱmolecule and retrovirus gene products may playimportantrole in the development of SLE.Atthe sametime,other molecules may also berelatedto SLE.