摘要
从鸡新城疫病毒刺激的鸭脾脏淋巴细胞培养液中提取基因组RNA,采用RT-PCR方法扩增鸭IL-18基因,插入pMD18-T载体,进行序列测定与分析。将成熟鸭IL-18基因亚克隆到表达载体PBCX中,并转化入大肠杆菌E.coliBL21进行表达。测序结果表明,该基因由603个核苷酸组成,包含一个完整阅读框,共编码201个氨基酸。表达载体经IPTG诱导表达出相对分子量约为59KD的融合蛋白。
Duck Interleukin-18 （DuIL-18） gene was amplified from chicken NDV-stimulated spleen lymphocytes by the method of RT-PCR The PCR product was cloned into the pMDI 8-T vector for sequencing and nucleic acid analysis. A prokaryotic expression plasmid, pBCX -ChiL18 was obtained by subcloning the encoding region of the DuIL-18 mature peptide into pBCX. Sequencing analysis showed that the nucleotide sequence of this DuIL-18 mature protein gene was 603bp long, including a complete ORF which encoded 201 amino acids. The recombinant DuIL- 18 gene was expressed efficiently in pBCX-DuIL- 18 plasmid transformed Escherichia coli strain （BL21） after induced by IPTG, the relative molecular weight ofthe mature peptide protein was approximately 59KD.
出处
《广东畜牧兽医科技》
2009年第2期28-30,37,共4页
Guangdong Journal of Animal and Veterinary Science
