| 小鼠白细胞介素-12基因在乳酸乳球菌中的表达 |
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| 引用本文： | 周强,周巧梅,李云,庞庆丰,曾因明.小鼠白细胞介素-12基因在乳酸乳球菌中的表达[J].徐州医学院学报,2007,27(3):155-158. |
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| 作者姓名： | 周强 周巧梅 李云 庞庆丰 曾因明 |
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| 作者单位： | 徐州医学院江苏省麻醉学重点实验室,江苏,徐州,221002 |
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| 摘 要： | 目的 在乳酸乳球菌中表达小鼠白细胞介素-12基因.方法 用Nsi Ⅰ和EcoR Ⅰ双酶切pGEM-IL-12质粒后,与经同样酶切的含有乳链杆菌肽A(nisA)启动子的pSEC-E7质粒连接,从而构建成pSEC-IL-12质粒;经电击转化,将重组质粒转入乳酸乳球菌NZ9000中,转化子在含有氯霉素的脑心浸液培养基上培养.用乳链杆菌肽(nisin)诱导白细胞介素-12表达,SDS-PAGE、Western blot鉴定表达产物;用酶联免疫吸附测定(ELISA)检测IL-12的量.结果 建立了白细胞介素-12乳酸菌基因表达系统.在nisin的诱导下,NZ9000能够产生并分泌白细胞介素-12约80 ng/L.结论 乳酸乳球菌能够表达一定数量的小鼠白细胞介素-12.
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| 关 键 词： | 白细胞介素-12 基因克隆 乳酸乳球 |
| 文章编号： | 1000-2065(2007)03-0155-04 |
| 修稿时间： | 2006年11月16 |
Expression of murine interleukin 12 gene in Lactococcus lactis |
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| ZHOU Qiang,ZHOU Qiao-mei,LI Yun,PANG Qing-feng,ZENG Yin-ming.Expression of murine interleukin 12 gene in Lactococcus lactis[J].Acta Academiae Medicinae Xuzhou,2007,27(3):155-158. |
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| Authors: | ZHOU Qiang ZHOU Qiao-mei LI Yun PANG Qing-feng ZENG Yin-ming |
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| Abstract: | Objective To express murine interleukin 12 gene in Lactococcus lactis. Methods The murine interleukin 12 gene from pGEM-IL-12 plasmid was digested with restriction endonuclease and was ligated with plasmid pSEC. Then the recombinant plasmid was transformed into Lactococcus lactis NZ9000 by electroporation. The expression of murine interleukin 12 gene induced by nisin was identified by SDS-PAGE and Western blot. The quantity of interleukin 12 was assayed by ELISA. Results Interleukin 12 gene expression system in Lactococcus lactis was established. Under nisin inducement, NZ9000 could produce and secrete interleukin 12 at a level of about 80 ng/L. Conclusion Certain amounts of murine interleukin 12 could be expressed in Lactococcus lactis. |
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| Keywords: | interleukin 12 gene cloning Lactococcus lactis |
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