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山东大学学报(医学版) ›› 2011, Vol. 49 ›› Issue (7): 39-43.

• 论文 • 上一篇    下一篇

人TIPE2基因启动子的鉴定及其表达调控区域的研究

陈洁1,宋永红2,王淑荣2,韩振龙3,姜学兵1,李国盛1,郭春1,张利宁1,石永玉1   

  1. 1.山东大学医学院免疫学研究所, 济南 250012; 2.山东省血液中心, 济南 250014;
    3.山东省警官总医院检验科, 济南 250002
  • 收稿日期:2011-01-29 出版日期:2011-07-10 发布日期:2011-07-10
  • 通讯作者: 石永玉(1970- ),男,副教授,硕导,主要从事肿瘤免疫及分子免疫研究。 E-mail:shiyongyu@sdu.edu.cn 张利宁(1960- ),女,教授,主要从事免疫调节及疾病的研究。E-mail:immuno@sdu.edu.cn
  • 作者简介:陈洁(1985- ),女,硕士研究生,主要从事分子免疫及肿瘤免疫的研究。 E-mail:ferradire@126.com
  • 基金资助:

    山东省中青年科学家科研奖励基金资助项目 (BS2009YY002);国家自然科学基金资助项目 (81071705,30500591)。

Identification of human TIPE2 promoter and its regulatory regions

CHEN Jie1, SONG Yong-hong2, WANG Shu-rong2, HAN Zhen-long3, JIANG Xue-bing1,  LI Guo-sheng1, GUO Chun1, ZHANG Li-ning1, SHI Yong-yu1   

  1. 1. Institute of Immunology, School of Medicine, Shandong University, Jinan 250012, China;
    2. Blood Center of Shandong Province, Jinan 250014, China;
    3. Clinical Laboratory,Shandong Police General Hospital, Jinan 250002, China
  • Received:2011-01-29 Online:2011-07-10 Published:2011-07-10

摘要:

目的      克隆人TIPE2基因启动子区及一系列5′截短片段,并鉴定其DNA表达调控区域。方法      采用PCR技术从人基因组DNA中克隆扩增TIPE2基因5′上游1252bp启动子序列并对该片段5′端的截短,分别瞬时转染人卵巢癌来源细胞系SKOV3,通过双萤光素酶活性的检测鉴定其启动子活性及表达调控区域。结果     克隆扩增的8个TIPE2基因启动子截短片段经测序证明无误;TIPE2-pGL4-F1重组质粒双萤光素酶活性检测显示其具有明显的启动子活性,约为pGL4Basic的7.9倍;5′系列截短片段重组质粒的启动子活性检测显示,与1252bp片段相比,TIPE2上游-965~-593bp、-501~-390bp缺失,启动子活性分别升高6.38、 7.82倍;-593~-501bp的缺失,使-501~+79bp片段相对于-593~+79bp片段的启动子活性降低2.04倍。结论     人TIPE2基因上游1252bp区域有明显的启动子活性,该片段内存在2个潜在的负性调控区和1个潜在的正性调控区。

关键词: 基因,TIPE2;启动区(遗传学);萤光素酶;基因表达调控

Abstract:

Objective     To identify the promoter region of human tumor necrosis factor-α induced protein 8-like 2(TIPE2), and important regulatory regions of a 1331bp putative TIPE2 promoter. Methods      A potential 1331bp promoter region and 7 fragments of 5′ truncation upstream TIPE2 gene were amplified from human genomic DNA by PCR. Then they were inserted into the luciferase report gene vector pGL4-Basic and transfected into human SKOV3 cells. Relative luciferase activity was detected to determine promoting activities. Results      DNA sequencing results indicated that promoter fragments were correctly identified. Analysis of relative luciferase activity showed that the 1331bp presented a strong promoter activity, which was increased by 7.9fold compared with pGL4-Basic.  TIPE2 promoter activity was respectively increased by 6.38-fold and 7.32-fold after -965bp to -593bp and -501bp to -390bp were deleted, while promoter activity was decreased by 2.04-fold after -593bp to -501bp was deleted. Conclusion     The 1331bp region upstream TIPE2 gene has high promoter activity. Two potential negative and one positive regulatory regions were identified by 5′truncation of  the 1331bp promoter region.

Key words: Genes, TIPE2;Promoter regions(genetics); Luciferase; Gene expression regulation

中图分类号: 

  • Q344+.14
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