真核双基因表达载体pIRES-IL-24-ES的构建与鉴定

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

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. 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. 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/Abstract

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

韩建军1，胡三元1，智绪亭1，梁晓红2，薛德文3. 真核双基因表达载体pIRES-IL-24-ES的构建与鉴定[J]. 山东大学学报(医学版), 2010, 48(3): 78-82.

HAN Jian-jun 1,　HU San-yuan 1,　ZHI Xu-ting 1,　LIANG Xiao-hong 2,　XUE De-wen 3 . Construction and identification of the eukaryotic co-expression plasmid pIRES-IL-24-ES[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2010, 48(3): 78-82.

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