家蚕杆状病毒表达系统优化表达人表皮生长因子(hEGF)的研究  

Optimized Expression of Human Epidermal Growth Factor (hEGF) by Using Silkworm-Baculovirus Expression System

作　　者：曾小群 李跃东 吴小锋[1,2] Zeng Xiaoqun;Li Yuedong;Wu Xiaofeng(College of Animal Sciences,Zhejiang University,Hangzhou 310058,China;Key Laboratory of Sikworm and Bee Resource Utlization and Innovation of Zhejiang Province,Hangzhou 310058,China)

机构地区：[1]浙江大学动物科学学院,杭州310058 [2]浙江省蚕蜂资源利用与创新研究重点实验室,杭州310058

出　　处：《蚕业科学》2022年第4期317-323,共7页ACTA SERICOLOGICA SINICA

基　　金：浙江省自然科学基金项目(LZ20C170001);国家自然科学基金项目(31972619,32172793)。

摘　　要：人表皮生长因子(hEGF)因具有促进表皮细胞增殖活性而被广泛用于烧伤、烫伤等医学领域。根据杆状病毒基因密码子偏好性优化hEGF序列,通过串联表达、融合表达以及构建携带双启动子和增强序列的载体等系列优化技术,提高重组hEGF在家蚕细胞中的表达水平。结果表明,通过上述一系列的优化方式可显著提高hEGF基因在BmN细胞中的转录水平,其中优化后hEGF的转录水平提高了24倍;Western blot分析表明,串联3个hEGF基因序列的蛋白质表达量最高。Human epidermal growth factor(hEGF) has the activity of promoting epidermal cells proliferation and is widely applied in medical fields such as burns and scalds therapy. After optimizing the hEGF sequence based on the baculovirus gene codon preference, the expression level of recombinant hEGF in silkworm cells was improved by a series of optimization techniques such as tandem expression, fusion expression, and construction of vectors with dual promoters and enhancer sequences. The results indicated that this series of optimization methods can significantly increase the transcription level of hEGF in BmN cells, among which the transcription level of hEGF after optimization was increased by 24 times. Western blot analysis showed that the protein expression level of three optimized hEGF sequences in series was the highest.

关 键 词：家蚕杆状病毒表达系统 人表皮生长因子 优化表达 串联表达 双启动子 

分 类 号：Q786[生物学—分子生物学]