EOLA1基因突变体的克隆和鉴定

Cloning and identification of a splicing variant of EOLA1 gene

目的 对ECV3 0 4细胞株EOLA1基因进行克隆测序以及mRNA水平检测。方法 采用RT PCR扩增EOLA1基因片断 ,连接到T载体进行测序。结果 测序发现一种目的基因mRNA剪接突变体 ,其第 3外显子起始部分比Genebank数据库相应序列多出 19个碱基。RT PCR显示剪接突变体与野生型EOLA1基因具有不同的表达水平和LPS反应性。结论 发现一个EOLA1基因剪接突变体 。

Objective To clone and sequence endothelial overexpression lipopolysaccharide associated factor 1 (EOLA1) gene of ECV304 cell line and to detect its mRNA expression. Methods EOLA1 gene fragment amplified by RT PCR was ligated to recombinant T vector for sequencing. Results A novel mRNA splicing variant of EOLA1 was identified with 19 redundant base pairs at the beginning of the 3rd exon of the target sequence published in Genebank. RT PCR showed that the splicing variant was quite different from the wild type in expression of mRNA and reactivity to LPS. Conclusion A novel mRNA splicing variant of EOLA1 gene has been identified, but its biological function is unclear.