摘要
目的获得印度尼西亚食蟹猴的干扰素-γ基因,为常用实验猕猴干扰素-γ的基因工程生产奠定基础。方法根据GenBank上公布的恒河猴干扰素-γ基因序列设计特异性引物,从印度尼西亚食蟹猴的外周血液中分离单核淋巴细胞,利用Trizol试剂,提取淋巴细胞的总RNA,通过RT-PCR的方法获得干扰素-γ基因片段,并对该片段进行克隆、鉴定和序列分析。结果扩增到一498bp的目的片段,经序列测定证实为印度尼西亚食蟹猴的干扰素-γ基因,与恒河猴、人及狒狒的干扰素-γ基因相比,同源性分别为100%、96%、99%。结论常用的两种实验猕猴食蟹猴与恒河猴的干扰素-γ基因完全相同。
Objective To obtain the IFN-γgene of Indonesian Cynomolgus monkeys and provide a theoretical and material basis for the production of the recombinant IFN-γ. Method Based on the published nucleotide sequence of IFNγ gene of rhesus monkey in GenBank,a pair of RT-PCR primers were designed and synthesized. Total RNA,isolated from the peripheral blood mononuclear cells by Trizol,was used as template to amplify the IFN-γgene by RT-PCR. The gene was identified by endonuclease,PCR and DNA sequencing. Result A 498 bp DNA fragments were amplified. DNA sequencing confirmed that the fragment was IFN-γof Cynomolgus monkeys. Compared with rhesus monkeys,humans and baboons,the homology of nucleotide sequence of IFN-γgene were 100% ,96 % and 99 % respectively. Conclusion The IFN-γ gene of cynomolgus monkey was identical to that of rhesus monkey.
出处
《中国比较医学杂志》
CAS
2010年第10期3-5,共3页
Chinese Journal of Comparative Medicine
基金
国家自然基金(30970416)
国家科技部基础平台项目(2005DKA21500)
