›› 2004, Vol. 47 ›› Issue (6): 837-843.

• 研究论文 • 上一篇    下一篇

重组昆虫杆状病毒构建和筛选技术进展

曹翠平,吴小锋   

  • 出版日期:2004-12-20 发布日期:2004-12-20

Progress in the techniques for construction and screening of recombinant insect baculovirus

CAO Cui-Ping, WU Xiao-Feng   

  • Online:2004-12-20 Published:2004-12-20

摘要: 昆虫杆状病毒作为高效的真核表达载体,现已广泛应用于各种外源目的基因的表达。但由于重组病毒产生的比例很低(通常只有0.1%~1%),成为制约该系统应用的技术瓶颈。本文概括了近年来发展的重组病毒的构建和筛选方法,主要介绍了杆状病毒的线性化技术和利用大肠杆菌-昆虫细胞穿梭载体构建并筛选重组杆状病毒的技术进展。

关键词: 重组昆虫杆状病毒, 空斑纯化, 线性化技术, Bac_to_Bac

Abstract: Using insect baculovirus as expression vector, various proteins have been produced successfully. But the frequency of recombination was very low (0.1%-1%) and it limited further application of the baculovirus expression vector system (BEVS). Many improved methods for the construction and isolation of recombinant baculo virus have been developed and are introduced here, with emphasis on linearization of baculovirus DNA and a method based on site-specific transposition of an expression cassette into a baculovirus shuttle vector propagated in E.coli.

Key words: Recombinant baculovirus, plaque assay, linearization technique, Bac-to-Bac