| 猪白细胞介素18成熟蛋白基因的克隆及在大肠杆菌中的表达 |
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| 引用本文： | 郑敏,金宁一,张洪勇,李昌,尹革芬.猪白细胞介素18成熟蛋白基因的克隆及在大肠杆菌中的表达[J].中国兽医学报,2003,23(5):430-432. |
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| 作者姓名： | 郑敏 金宁一 张洪勇 李昌 尹革芬 |
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| 作者单位： | 解放军军需大学,军事兽医研究所,吉林,长春,130062 |
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| 基金项目： | 国家 8 63计划资助项目 (2 0 0 1AA2 13 0 71) |
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| 摘 要： | 通过RT—PCR方法从用PHA和LPS刺激的猪脾脏细胞中扩增出猪白细胞介素18(pIL-18)成熟蛋白基因的cDNA，克隆到T载体pUCm-T后，序列测定表明，pIL-18成熟蛋白基因核苷酸长度为474bp，编码157个氨基酸。将该基因片段克隆到大肠杆菌表达载体pET-28a构建重组质粒pETIL18m，转化大肠杆菌BL21(DE3)，并用IPTG诱导。重组菌菌体裂解物SDS—PAGE可检测到相对分子质量为19000的重组蛋白，免疫印迹法证实该重组蛋白可以与人IL-18单抗发生特异性免疫反应。
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| 关 键 词： | 猪白细胞介素18 成熟蛋白 基因克隆 原核表达 RT-PCR 大肠杆菌 免疫佐剂 免疫调节剂 |
| 文章编号： | 1005-4545(2003)05-0430-03 |
| 修稿时间： | 2002年11月28 |
Cloning and Expression of Porcine Interleukin-18 Mature Protein Gene in Escherichia coli |
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| ZHENG Min,JIN Ning yi ,ZHANG Hong yong,LI Chang,YIN Ge fen.Cloning and Expression of Porcine Interleukin-18 Mature Protein Gene in Escherichia coli[J].Chinese Journal of Veterinary Science,2003,23(5):430-432. |
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| Authors: | ZHENG Min JIN Ning yi ZHANG Hong yong LI Chang YIN Ge fen |
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| Institution: | ZHENG Min,JIN Ning yi *,ZHANG Hong yong,LI Chang,YIN Ge fen |
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| Abstract: | Porcine interleukin 18 mature protein gene was amplified from porcine spleen cells stimulated with phytohemagglutinin(PHA) and lipopolysaccharide(LPS). PCR product was cloned into the T vector pUCm T for sequencing. The result showed the nucleotide sequence of this gene was 474 bp. Then, this pIL 18 mature protein gene was subcloned into the prokaryotic expressing plasmid vector pET 28a and transformed into host E.coli strain BL21(DE3) for expression. The expression of pIL 18 mature protein gene was identified by SDS PAGE and Western blotting. The results revealed it had a molecular weight of 19 000, and could be specifically recognized by the mouse monoantibody to human IL 18. |
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| Keywords: | porcine interleukin 18 mature protein cloning prokaryotic expression |
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