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The study on regulatory function of the cis-regulating elements of ARE and E-box in the human GCLC gene
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
- Issue:
- 3
- DOI:
- 10.3760/cma.j.issn.1674-1927.2007.03.003
- Key Word:
- 基因转录调控;转录调控元件ARE;转录调控元件E-box;诱变
Abstract： 目的 通过对人γ-谷氨酰半胱氨酸酶催化亚单位(GCLC)基因调控区的ARE及E-box顺式调控元件诱导缺失突变,研究它们在人支气管上皮细胞内对GCLC基因转录调控功能的影响.方法 应用PCR方法克隆人GCLC基因调控区的大部分核苷酸序列,构建表达虫荧光素酶报告基因的野生型质粒PL45.采用PCR重叠延伸产生特异位点诱变的方法对ARE及E-box顺式调控元件进行缺失突变,构建相应的突变型质粒PLdel-ARE及PLdel- E-box.通过瞬时基因转染方法将上述质粒转染人支气管上皮细胞系16HBE,观察基因突变后的基因转录调控功能.结果 质粒PL45的虫荧光素酶相对活性值为9949.75±1441.33、质粒PLdel-ARE的虫荧光素酶相对活性值为21258.75±1563.64、质粒PLdel-E-box的虫荧光素酶相对活性值为17082.00±89.51.与野生型质粒PL45相比,PLdel-ARE、PLdel-E-box突变型质粒表达的虫荧光酶相对活性值显著上升.结论 ARE及E-box顺式调控元件具有负性调控作用.
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