摘要
目的:通过克隆人的Interleukin-18基因,构建Interleukin-18基因的哺乳动物细胞真核表达载体。方法:把人基因组DNA通过PCR获得Interleukin-18,克隆载体pMD18-T/Interleukin-18,并将其转化到大肠杆菌里,最后进行重组真核表达载体pCDNA3.1/Interleukin-18的构建和鉴定。结果:以人总DNA为模板扩增出580 bp左右的特异性条带,测序结果与GeneBank测序结果相比,除两个碱基外,其余的完全一致,翻译成的氨基酸序列相同。对重组质粒pCDNA3.1/Interleukin-18进行双酶切鉴定并测序,结果也完全一致。结论:重组质粒pCDNA3.1/Interleukin-18由真核载体pCDNA3.1和Interleukin-18片段组成。且插入的Interleukin-18片段与已知序列完全相同。
Objective The eukaryotic expression vector of interleukin-18 was constructed according to the interleukin-18 gene of human.Method Interleukin-18 gene was obtained from human ge nome DNA by PCR.Vector pMD18-T/interleukin-18 was cloned and transformed into Escherichia coli,and the recombinant pCDNA3.1/interleukin-18 was constructe d and identified. Results The result showed that 580 bp specific fragment was amplified and identified.Sequencing result showed that the reading frame was not changed except one base compared with the GeneBank,and amino acid sequence was the same. Conclusion The result of i dentification and sequencing of pCDNA3.1 /interleukin-18 plasmid is the accordance with the expected length.
出处
《吉林医学》
CAS
2011年第22期4517-4519,共3页
Jilin Medical Journal
基金
吉林省科技发展计划项目[项目编号:200905176]
