摘要
目的通过白细胞介素-6(IL-6)相关新基因的生物学分析,研究IL-6的作用机制。方法利用IL-6相关表达序列标签(EST)进行电子克隆获得924 bp全长新基因,后采用RT-PCR方法从IL-6激活的人U937细胞所提的总RNA中钓取,此片断连到pGEM-Teasy质粒上并测序。结果成功钓取了IL-6相关EST电子克隆的全长cDNA基因。结论使用电子克隆技术对于发现新基因有重要的指导意义。
Objective To understand the action mechanism of interleukin (IL)-6 through investigation of its related genes. Methods Using electronic cloning with IL-6-related expressed sequence tags (ESTs), a novel gene at its full length of 924 bp was acquired. Reverse transcriptase-PCR method was subsequently employed to amplify the cDNA of this gene from the total RNA of human U937 cells, which were previously activated with 100 ng/ml IL-6 for 8 h. The fragment was then ligated into the pGEM-Teasy vector and sequence analysis was conducted. Result The whole cDNA gene was successfully fished. Conclusion Electronic cloning technology can be instrumental in identifying novel genes.
出处
《第一军医大学学报》
CSCD
北大核心
2003年第1期34-37,共4页
Journal of First Military Medical University
基金
国家杰出青年科学基金(39925019)~
关键词
白细胞介素-6
相关新基因
生物学分析
interleukin-6
U937 cells
expressed sequence tags
electronic cloning