具有增强蛋白质表达活性序列ER3的筛选及其功能区域的初步鉴定

Screening of ER3 Sequences Enhanced Protein Expression Activity and Its Functional Regions Identification

目的：利用构建的增强子样序列筛选载体,筛选在大肠杆菌中增强外源蛋白质表达的序列,并利用删除突变体初步鉴定其功能区域.方法：以氯霉素乙酰转移酶（CAT）基因序列与人乳头瘤病毒（HPV）主要衣壳蛋白基因 L1 的截短序列L11连接作为报告基因,从采集的样品中筛选增强子样序列,通过蛋白质表达来检测其增强活性,并通过构建删除突变体来初步鉴定其功能区域.结果： 成功筛选到一条增强子样序列,可使检测载体氯霉素抗体提高11倍,融合蛋白表达水平提高2.26倍,其功能区域主要集中在1~265bp.结论：从收集的样品中成功筛选出一个增强子序列,能提高外源基因在大肠杆菌中的表达.

Objective： Using a constructed enhancer sequence detecting vector, screening sequences that could enhanced heterologous protein expression in Escherichia coli, and preliminary identified its functional domain by deletion mutants. Methods： Chloramphenicol acetyltransferase （CAT） gene and truncated human papillomavirus （HPV） major capsid protein gene L1 （named L11） were fusion expressed as a report gene; to screen enhancer-like sequence from the collected samples, and detected the enhanced activity by protein expression. The functional regions were identified by construction of deletion mutants. Results： One enhancer-like sequence was successfully screened, which enhanced 11-folds of chloramphenicol resistance; fusion protein expression level increased 2.26-folds; the functional areas are located in the 1~265bp. Conclusion： An enhancer-like sequence were successfully screened, which can improve the expression of heterologous gene in Escherichia coli.