摘要
目的克隆小鼠白细胞介素1cDNA(IL1cDNA),构建小鼠IL1cDNA逆转录病毒载体。方法利用逆转录聚合酶链反应(RTPCR),从小鼠胸腺细胞中扩增出小鼠IL1cDNA,克隆到测序载体pBluscript中,测序后,再亚克隆到逆转录病毒载体pLNCX中。结果RTPCR扩增出约518bp片段,经酶切鉴定显示小鼠IL1cDNA逆转录病毒载体(IL1pLNCX)构建成功。结论小鼠IL1cDNA与已发表的其他种小鼠的序列相同,IL1cDNA可能无种间差异。为进一步研究IL1在中枢神经系统病理状态下的作用机制提供了有利的研究工具。
Objective:To clone the cDNA of mouse interleukin-1 and su bc lone the cDNA into retroviral vector pLNCX as a tool for the purpose of elucidat ing the role of IL-1 in central nerve system’s disorder.Methods:The mouse IL-1 cDNA was obtained from mouse cerebral cortical cells by RT-PCR.The cDNA wa s first cloned into pBluscript to facilitate the sequencing and then subcloned i nto the retroviral vector pLNCX.Results:A 518 bp DNA fragment was obtained by RT-PCR and the insertion of the mouse IL-1 cDNA into pLNCX was confirmed by d ouble digestion with BamH1 and Xho1. The sequencing results was identified.Concl usion:The sequence of the SD mouse IL-1 cDNA is consistent with that of other strains of mouse,and the mouse IL-1 cDNA of different strains may have the same cDNA sequence.The present experiment demonstrates a successful cloning of mouse IL-1 cDNA into the retroviral vector pLNCX.
出处
《实用临床医学（江西）》
CAS
2005年第7期20-22,共3页
Practical Clinical Medicine
