Construction of pEgr-ssEndostatin and mouse endostatin expression in vitro induced by ionizing radiation

LI Xiujuan; LI Zhiguo; TIAN Mei; YANG Wei; LIU Linlin; LI Xiuyi

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Construction of pEgr-ssEndostatin and mouse endostatin expression in vitro induced by ionizing radiation

更新时间：2023-03-30

- Construction of pEgr-ssEndostatin and mouse endostatin expression in vitro induced by ionizing radiation
- Journal of Radiation Research and Radiation Processing Vol. 22, Issue 1, Pages: 35-38(2004)
- 作者机构：
  
  1.河南大学医学院细胞与分子免疫实验室　开封　475001
  2.吉林大学卫生部放射生物学重点实验室　长春　130021
  3.吉林大学卫生统计教研室　长春　130021
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LI Xiujuan, LI Zhiguo, TIAN Mei, et al. Construction of pEgr-ssEndostatin and mouse endostatin expression in vitro induced by ionizing radiation. [J]. Journal of Radiation Research and Radiation Processing 22(1):35-38(2004)
DOI：

LI Xiujuan, LI Zhiguo, TIAN Mei, et al. Construction of pEgr-ssEndostatin and mouse endostatin expression in vitro induced by ionizing radiation. [J]. Journal of Radiation Research and Radiation Processing 22(1):35-38(2004) DOI：

摘要

为探讨pEgr−ssEndostatin重组质粒转染B16细胞后接受不同辐射剂量以及接受同一剂量不同时程endostatin表达的规律。采用RT−PCR方法从鼠肝脏中扩增出endostatin cDNA，连入信号肽，构建了pEgr−ssEndostatin重组质粒，用脂质体介导的转染法转染小鼠B16细胞，用ELISA方法检测B16细胞上清中endostatin的表达水平。结果表明的分泌型endostatin序列与报道完全一致，Egr−1启动子和Endostatin cDNA正确插入表达载体pcDNA3.1；pEfr−ssEndostatin具有辐射诱导表达特性。提示小鼠endostatin基因成功地被克隆和表达，Egr−1启动子具有辐射激活和诱导下游基因表达增强的功能。

Abstract

To study the regularity of mouse endostatin expression in B16 cells transfected by pEgr−ssEndostatin after different doses of X-ray irradiation and different time after the irradiation, mouse endostatin cDNA was amplified by RT−PCR and signal peptide was ligated to it. The pEgr−ssEndostatin recombinant plasmid was constructed and transfected into B16 cells with liposome and cells were irradiated. The expression of endostatin in the supernatant medium of B16 cells was detected by ELISA. The results showed that the cDNA sequence of mouse secretable endostatin is identical to the literatures. Egr−1 promoter and secretable endostatin cDNA were inserted into expression vectors correctly. pEgr−ssEndostatin had the expression property induced by radiation. It suggests that the Egr−1 promoter can be activated by ionizing irradiation and expression of downstream gene can be regulated.

关键词

内皮抑素cDNAEgr−1启动子pEgr−ssEndostatin重组质粒辐射诱导表达

Keywords

Mouse endostatin cDNAEgr−1 promoterRecombinant plasmid of pEgr−ssEndostatin expression induced by radiation

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Department of Biology, Shantou University Medical college

MH Radiobiology Research Unit, School of Preventive Medicine Jilin University

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