严重烧伤F344大鼠免疫细胞中一个EST全长序列的扩增

目的:探讨免疫细胞在大面积烧伤后全身免疫功能紊乱中的变化,对F344大鼠烧伤后血液淋巴细胞和单核细胞中的1个差异表达基因片段(AI764697.1)进一步研究。方法:根据该片段核苷酸序列设计双向引物和巢式引物,应用cDNA末端快速扩增技术(SMARTRACE)对该片段进行双向扩增,扩增后结果经测序,用Northernblot杂交验证。结果:设计的两对引物经扩增都得到了产物,克隆测序后得到1条长度为592bp的片段。Northernblot杂交在烧伤后得到1个长度约700bp的产物,在脾脏中表达水平较高,与RACE结果符合。该核苷酸序列已得到GenBank登录号AF244895。结论:在严重烧伤F344大鼠的血液淋巴细胞和单核细胞中分离并扩增出一个新的基因全长序列,其来源、定位和功能需要进一步研究。

Whole sequence amplification of an expressed sequence tag in immune cells of F344 rats with severe burns

AIM: To examine the changes in immune cells which might be involved in systemic immune dysfunction after severe burns, we detected the sequence of expressed sequence tag in lymphocytes and monocytes of F344 rats post-burn. METHODS: Two pairs of primers were designed depending on the sequence of AI764697.1. SMART RACE technology was employed for amplification of whole sequence. Furthermore, the amplified sequence was testified by Northern blot. RESULTS: A 592bp sequence, which was testified by Northern blot, was suceessfally obtained in our experiments. This product was accepted by GenBank and its accession number is AF244895. CONCLUSION: A cDNA whole sequence in immune cells of F344 rats after severe burns was amplified successfully. More research work should be done to identify the resource, location in genome and function of this product.