人白细胞介素-18真核表达载体的构建及测序分析

Construction and sequencing of human interleukin-18 eukaryotic expression vector

目的 :构建含有人白细胞介素 18(IL 18)基因的真核表达载体 ,并对其进行测序分析。 方法 :通过分子克隆技术 ,将IL 18基因插入真核表达载体 pVAX1中 ,构建真核表达载体pVAX1 IL 18,并经BamHⅠ /EcoRⅠ双酶切及测序鉴定。利用Blast程序 ,搜索NCBIGenBank中与重组质粒 (pVAX1 IL 18)中编码IL 18基因同源的序列。结果 :人IL 18基因成功地克隆至真核表达载体pVAX1中 ,pVAX1 IL 18中编码IL 18的序列与GenBank中所报道的IL 18编码序列完全一致。 结论 :成功地构建人IL 18基因真核表达载体 。

Objective:To construct the eukaryotic expression vector of interleukin-18 gene and carry out analysis of pVAX1-IL-18 sequence . Methods:The recombinant eukaryotic expression vector pVAX1-IL-18 was constructed by inserting interleukin-18 gene into the eukaryotic expression vector pVAX1 with molecular cloning technique . It was confirmed by restrictive enzymes(BamHⅠ/ EcoRⅠ) digestion and analysis of DNA sequencing . Similar nucleotide sequences encoding IL-18 of pVAX1-IL-18 were looked up by Blast means in NCBI GenBank. Results:Restrictive enzymes digestion analysis and DNA sequencing results revealed that the interleukin-18 gene was cloned into the eukaryotic expression vector pVAX1 successfully .The sequence of encoding IL-18 of pVAX1-IL-18 was exactly the same as the reported sequence of encoding human IL-18 in GenBank . Conclusion: The recombinant eukaryotic expression vector pVAX1-IL-18 has been constructed successfully, which lay the foundation for pVAX1-IL-18 as a genetic adjuvant of DNA vaccine.