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山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (3): 78-82.

• 论文 • 上一篇    下一篇

真核双基因表达载体pIRES-IL-24-ES的构建与鉴定

韩建军1,胡三元1,智绪亭1,梁晓红2,薛德文3   

  1. 1. 山东大学齐鲁医院普通外科, 济南 250012;2. 山东大学医学院免疫学实验室, 济南 250012;
    3. 山东省肿瘤医院肿瘤介入治疗中心, 济南 250117
  • 收稿日期:2009-12-16 出版日期:2010-03-16 发布日期:2010-03-16
  • 通讯作者: 胡三元(1962- ),男,教授,博士生导师,主要从事普外科肿瘤的研究。
  • 作者简介:韩建军(1975- ),男,博士研究生,主治医师,主要从事肿瘤外科临床与基础研究。
  • 基金资助:

    山东省自然科学基金项目(Y2007C150)

Construction and identification of the eukaryotic co-expression plasmid pIRES-IL-24-ES

HAN Jian-jun 1, HU San-yuan 1, ZHI Xu-ting 1, LIANG Xiao-hong 2, XUE De-wen 3   

  1. 1. Department of General Surgery, Qilu Hospital of Shandong Univeristy, Jinan 250012, China; 
    2. Department of Immunology, School of Medicine, Shandong Univeristy, Jinan 250012, China;
    3. Department of Tumor Interventional Therapy, Shandong Tumor Hospital, Jinan 250117, China
  • Received:2009-12-16 Online:2010-03-16 Published:2010-03-16

摘要:

目的    构建白介素24(IL-24)与内皮抑素(ES)的真核双基因表达载体,检测它们在体外的表达。方法    从外周血和人胎肝组织中经 RT-PCR分别扩增IL-24与ES cDNA序列,并将其定向克隆至真核双基因表达质粒pIRES中,重组质粒pIRES-IL-24-ES,经酶切及测序鉴定后,转染NIH3T3 成纤维细胞。经RT-PCR及ELISA法检测IL-24和ES在NIH3T3中的表达。结果    重组真核双基因表达载体pIRES-IL-24-ES构建成功,IL-24与ES可在NIH3T3细胞中有效表达。结论    真核双基因表达载体pIRES-IL-24-ES成功构建为研究肿瘤的基因治疗提供了一定线索。

关键词: 白介素24;内皮抑素;分泌表达载体;肿瘤

Abstract:

Objective    To construct the eukaryotic co-expression plasmid encoding interleukin-24(IL-24) and endostatin(ES), and to detect expression of the plasmid in vitro. Methods    The cDNA encoding Il-24 and ES were obtained by RT-PCR amplifications from peripheral blood and normal fetus liver respectively and cloned into an eukaryotic expression plasmid internal ribosome entry site(pIRES). The co-nstructed plasmid pIRES-IL-24-ES was confirmed by restriction enzymolysis and DNA sequencing. Fibroblast NIH3T3 cells were transformed by plasmid pIRES-IL-24-ES. RT-PCR and ELISA were used to identify the coexpression of IL-24 and ES in NIH3T3 cells. Results    The eukaryotic expression plasmid pIRES-IL-24-ES was  successfully constructed and the plasmid could  efficiently co-express IL-24 and ES in NIH3T3 cells. Conclusion    The successful construction of eukaryotic expression plasmid pIRES-IL-24-ES establishes an experimental basis for cancer treatment.

Key words: Interleukin-24; Endostatin; Expression vector; Carcinoma

中图分类号: 

  • R392.1
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