颗粒裂解肽抗菌结构域在大肠杆菌中表达水平的分析.pdfVIP

27 3 Vol27 No3 2010 6 JOURNAL OF BIOLOGY Jun, 2010 doi10. 3969 /jissn1008- 9632. 2010030 8 , ( 安徽大学生命科学学院, 合肥 230039) : 为实现颗粒裂解肽抗菌结构域的高效表达并避免 对宿主菌的毒性, 研究设计了共表达阴离子配体重组生 产阳离子抗菌肽的方法结果证明该方法可有效提高阳离子抗菌肽的表达产量同时研究分析了颗粒裂解肽不同 抗菌结构域在同一原核表达载体 pACYCDuet1中表达水平的实验数据通过计算机程序对mRNA 二级结构的模 拟分析, 计算并比较出 二级结构生成自由能之间的差异, 提出了预测颗粒裂解肽抗菌结构域在 E. coli 中表达水平 的几点依据, 参考这些依据可以对不同抗菌肽基因进行改造以获得高效表达 : 阳离子抗菌肽; 阴离子配体; 共表达; 二级结构; 翻译起始区 : Q786 : A : 1008- 9632( 2010) 03- 00 8- 05 Analysis of the level of expression of antim icrobial domain s of granulysin in E. coli ZHOU Peng, ZHA X iangdong ( School of Life ScienceAnhu iUniversity, H efei230039, China) A bstrac t: In order to exp lore a new approach for high expression of the d ifferent antmi icrobial dom ains of granulysin and to avoid the tox ic effects onE scher ich ia co li host cells during its expression, a novel strategy for the production of recom binant cationic antmi icrobial pep tidesw as designed by coexpressing an anion ic ligand. The results dem onstrated that a h igher expression yield w as achieved by using the coexpress ion m ethod. Som e expermi ental data abou t the expression levels of the dom ains in vector pACYCDuet1 w ere also analyzed. The mRNA secondary structuresw ere pred icted s mi u ltaneously by compu teraided design. Resu lts of the predictive analysis show ed the difference between the energy of mRNA secondary structures. A ccording to som e references, w e suggest a set of judge m ents. It w ill be helpfu l for us to m odify these antmi icrob ial dom ains before exper