Laboratory of Genetics and Molecular Biology, Northeast Forestry University, Harbin 150040, China.
Anal Biochem. 2010 Oct 15;405(2):272-4. doi: 10.1016/j.ab.2010.06.031. Epub 2010 Jun 20.
As a novel selection marker for DNA cloning, the genetically encoded photosensitizer KillerRed was used to achieve red/white cloning screening within the pZK18T T-vector system. KillerRed functioned without cofactors, inducers, or substrates. KillerRed-based red/white cloning screening was reliable in that bacteria containing DNA inserts that disrupt functional KillerRed expression form white colonies or red colonies as backgrounds. KillerRed simplifies assembly of customized vector and recombinant screening procedures. No special host or culture medium is required to amplify and prepare the vector in larger quantities. It makes high-throughput general-purpose cloning screening simpler, less expensive, and more effective.
作为一种新型的 DNA 克隆选择标记,遗传编码的光敏剂 KillerRed 被用于在 pZK18T T-载体系统中实现红色/白色克隆筛选。KillerRed 无需辅因子、诱导剂或底物即可发挥作用。基于 KillerRed 的红色/白色克隆筛选是可靠的,因为含有破坏功能性 KillerRed 表达的 DNA 插入物的细菌形成白色或红色菌落作为背景。KillerRed 简化了定制载体的组装和重组筛选程序。不需要特殊的宿主或培养基来大量扩增和准备载体。它使高通量通用克隆筛选更简单、更经济、更有效。