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α_1-酸性糖蛋白(α_1-AGP)是由肝脏合成的一个糖蛋白,固醇类激素及急性期炎症对其基因表达均具有正调控效应,且此正调控效应在转录水平发挥作用.为进一步研究α_1-AGP基因表达调控机理,作者以α_1-AGP cDNA为探针,从大鼠肝基因文库中分离一长度约15.5 kb的DNA片段,经内切酶图谱及部分序列测定均与Reinke等报道相同,含6个外显子及两侧侧翼区域. 参照Gorski等方法从成年(65d)大鼠肝脏制备可溶性核抽提物,以腺病毒基因为模板,用8mol/L-5％聚丙烯酰胺凝胶电泳监测所制备的核抽提物的转录活性.结果表明该抽提物适合于体外转录系统研究.以具有不同长度的5’-侧翼序列的α_1-AGP基因片段为模板,即EB_2-5.02 kb至+380bp(+380 bp
Alpha 1-acid glycoprotein (α 1 -AGP) is a glycoprotein synthesized by the liver. Steroid hormones and acute phase inflammation have a positive regulatory effect on the gene expression, and this positive regulatory effect plays a role in the transcriptional level. To study the regulation mechanism of α_1 -AGP gene expression, a 15.5 kb DNA fragment was isolated from rat liver gene library by using α_1 -AGP cDNA as a probe. Both endonuclease map and partial sequence analysis were performed by Reinke et al. , Containing 6 exons and flanking region on both sides.According to Gorski and other methods from adult (65d) rat liver preparation of soluble nuclear extract, adenovirus gene as a template, with 8mol / L-5% polyacrylamide Gel electrophoresis was used to monitor the transcriptional activity of the prepared nuclear extracts.The results showed that the extract was suitable for in vitro transcription system study.The α_1-AGP gene fragments with different length 5’-flanking sequences were used as templates, that is, EB_2- 5.02 kb to + 380 bp (+380 bp