甘氨酸和TritonX-100对ZZ-EGFP融合蛋白分泌表达影响的初步研究  

Effect of glycine and Triton X-100 on the secretion of ZZ-EGFP fusion protein

作　　者：唐金宝[1] 杨洪鸣[2] 张义军[2] 李万忠[1] 

机构地区：[1]潍坊医学院药学教研室,261042 [2]潍坊医学院药理学教研室,261042

出　　处：《中国医药生物技术》2007年第4期266-269,共4页Chinese Medicinal Biotechnology

摘　　要：目的考察不同浓度的甘氨酸和TritonX-100对葡萄球菌蛋白质AZZ亲和肽-增强型绿色荧光蛋白(ZZ-EGFP)融合蛋白在大肠杆菌分泌表达的影响。方法研究设计为两因素三水平析因设计。向液体培养基中分别加入终浓度0、1%、2%的甘氨酸和0、1%、2%的TritonX-100(共9种组合方式,终浓度均为0者为对照组),诱导大肠杆菌周质腔内ZZ-EGFP融合蛋白泄漏到液体培养基中,以培养上清液荧光强度为观察指标,通过ZZ-EGFP融合蛋白浓度-荧光强度标准曲线快速检测培养基中ZZ-EGFP融合蛋白表达量。结果培养基中分别加入1%、2%甘氨酸或1%、2%TritonX-100培养后,培养上清液荧光强度分别为283±11、711±19和622±25、733±25,与对照组(74±5)比较,组间差异均有统计学意义(甘氨酸:F=10.881,P=0.024;TritonX-10:F=12.848,P=0.018);而且甘氨酸与TritonX-100两者之间有交互效应(F=5.441,P=0.005),其中培养基中同时含有终浓度2%甘氨酸及1%TritonX-100时培养上清液荧光强度最高(1854±45),ZZ-EGFP融合蛋白分泌表达量达到10.4mg/L,与对照组(0.94mg/L)相比提高了11倍。结论甘氨酸和TritonX-100能提高ZZ-EGFP融合蛋白在液体培养基中的分泌表达量。Objective To study the effect of glycine and Triton X-100 on the secretion of ZZ-EGFP fusion protein from E.coli into the culture medium. Methods A two-factor three-level experimental design was applied. Glycerin and Triton X-100 at final concentrations of 0, 1%, and 2% was added into liquid culture medium (9 groups in total; in the control group, both the concentrations of glycerin and Triton X-100 were 0) to induce the secretion of ZZ-EGFP fusion protein from E.coli. The fluorescent intensity of the supernatant was detected to determine the level of ZZ-EGFP fusion protein expression in the culture medium by using a concentration-fluorescent intensity standard curve. Results The fluorescent intensity was 283 ± 11 in that with 1% glycerin, 711 ± 19 in that with 2% glycerin, 622 ± 25 in the medium added with 1% Triton X-100, and 733 ± 25 in that with 2% Triton X-100. All the results were significantly higher than that in the control group (74 ± 5; F = 10.881, P = 0.024 for glycerin and F = 12.848, P = 0.018 for Triton X-100). There were interaction between glycine and Triton X-100 (F = 5.441, P = 0.005). In the medium contained 2% glycine and 1% Triton X-100, the fluorescent intensity reached the highest level (1854 ± 45), and the level of ZZ-EGFP fusion protein expression was 10.4 mg/L, which was 11 times higher than that in the control group (0.94 mg/L). Conclusion Glycine and Triton X-100 can enhance the secretion of ZZ-EGFP fusion protein into the liquid culture medium.

关 键 词：甘氨酸 辛苯昔醇 重组融合蛋白质类 葡萄球菌蛋白质A 绿色荧光蛋白质类 大肠杆菌 

分 类 号：R341[医药卫生—基础医学]