抑制差减杂交筛选严重烫伤大鼠免疫细胞差异基因  

Screening of aberrant genes of immunocytes in severely scalded rats by inhibition of subtractive hybridization

作　　者：石胜军[1] 夏照帆[1] 

机构地区：[1]第二军医大学长海医院烧伤科

出　　处：《中华烧伤杂志》2001年第5期295-297,共3页Chinese Journal of Burns

基　　金：国家杰出青年科学基金资助项目 ( 3972 5 0 2 9)

摘　　要：目的　探索严重烧伤后早期免疫细胞基因表达变化与烧伤后免疫功能紊乱的关系。　方法　分离严重烫伤F34 4雌性大鼠的外周血免疫细胞并提取mRNA ,以烫伤前为对照进行抑制差减杂交筛选差异表达基因 ,将差异基因克隆后测序 ,与Genbank进行同源性比较。　结果　分离的免疫细胞经检测基本为单核细胞及淋巴细胞 ,抑制差减杂交得到多个位于 2 0 0～ 40 0bp的条带 ,将它们克隆到 pGEM -T质粒载体 ,构建了差异表达基因库。随机测序 2 0个克隆 ,得到的基因与烧伤后机体改变相符 ,部分新基因序列在Genbank登录。　结论　严重烧伤早期外周血免疫细胞的基因表达就有改变。构建了F34 4雌性大鼠的差异表达基因库 ,从中检出的部分基因可能对严重烧伤后的免疫功能紊乱有诱发作用。Objective To explore the relationship between the postburn changes of gene expression of immunocytes and post burn immunological functional disturbance in severely scalded rats. Methods Immunocytes of female F344 rats with severe sca lding were harvested and the mRNA was extracted for the inhibition of the expres sion of aberrant gene selected by subtractive hybridization in contrast to that before scalding.The aberrant genes were cloned and sequenced and compared to tho se in Genebank for homology. Results The harvested immunocytes were identified by LM to be monocytes and lymphocytes.Multiple strips located at 200～400 bp were obtained by inhibiting subtractive hybridization and were cloned into PGEM-T easy plasmid vector.Thus the genebank of aberrant expression was constructed.Twenty clones were sequenced randomly and partial obtained known genes were in accord with postburn systemic changes.Several novel genes were accepted and registered by Genebank . Conclusion There exhibited change in the gene expression of peri pheral immunocytes at early postburn stage in rats inflicted by severe burn.

关 键 词：烧伤 免疫细胞 差异表达基因 免疫功能 抑制差减杂交筛选法 

分 类 号：R644[医药卫生—外科学]