摘要
目的 克隆小鼠白细胞介素 - 18cDNA(mIL - 18cDNA) ,构建mIL - 18cDNA逆转录病毒载体。方法 利用逆转录聚合酶链反应 (RT -PCR)方法从昆明小鼠肝脏细胞中扩增出mIL - 18cDNA ,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 RT -PCR扩增出约 840bp片段 ,经酶切鉴定显示mIL - 18cDNA逆转录病毒载体 (mIL - 18pLNCX)构建成功。结论 昆明小鼠mIL - 18cDNA与已发表的其他种小鼠的序列相同 ,mIL - 18cDNA可能无种间差异。我们已成功地把mIL -
Objective To clone the cDNA of mouse interleukin-18 (mIL-18) and subclone the cDNA into retroviral vector pLNCX for the purpose of evolving gene therapy for cancers. Methods The mIL-18 cDNA was obtained from mouse hepatic cells by RT-PCR. The cDNA was first cloned into pBluscript to facilitate the sequencing and then subcloned into the retroviral vector pLNCX. Result An 840 bp DNA fragment was obtained by RT-PCR and the insertion of the mIL-18 cDNA into pLNCX was confirmed by double digestion with HindIII and ClaI. Conclusion The sequence of the Kunming mouse IL-18 cDNA was consistent with that of other strains of mouse, and the mIL-18 cDNA of different strains may have the same cDNA sequence. The present experiment demonstrates a successful cloning of mIL-18 cDNA into the retroviral vector pLNCX.
出处
《徐州医学院学报》
CAS
2001年第2期89-91,共3页
Acta Academiae Medicinae Xuzhou
