The successful production of recombinant protein for biochemical, biophysical, and structural biological studies critically depends on the correct expression organism. Currently, the most commonly used expression organisms for structural studies are Escherichia coli (~70% of all PDB structures) and the baculovirus/ insect cell expression system (~5% of all PDB structures). While insect cell expression is frequently successful for large eukaryotic proteins, it is relatively expensive and time‐consuming compared to E. coli expression. Frequently the decision to carry out a baculovirus project means restarting cloning from scratch. Here we describe an integrated system that allows simultaneous cloning into E. coli and baculovirus expression vectors using the same PCR products. The system offers a flexible array of N‐ and C‐terminal affinity, solubilization and utility tags, and the speed allows expression screening to be completed in E. coli, before carrying out time and cost‐intensive experiments in baculovirus. Importantly, we describe a means of rapidly generating polycistronic bacterial constructs based on the hugely successful biGBac system, making InteBac of particular interest for researchers working on recombinant protein complexes.

中文翻译：

---

InteBac-整合的细菌和杆状病毒表达载体套件。

用于生化，生物物理和结构生物学研究的重组蛋白的成功生产关键取决于正确的表达生物。当前，用于结构研究的最常用表达生物是大肠杆菌（约占所有PDB结构的70％）和杆状病毒/昆虫细胞表达系统（约占所有PDB结构的5％）。尽管昆虫细胞表达对于大型真核蛋白通常是成功的，但与大肠杆菌表达相比，它相对昂贵且费时。通常，执行杆状病毒项目的决定意味着从头开始重新克隆。在这里，我们描述了一个允许同时克隆到大肠杆菌中的集成系统和使用相同PCR产物的杆状病毒表达载体。该系统提供了灵活的N端和C端亲和力，增溶性和效用标签阵列，并且该速度允许在进行杆状病毒的时间和成本密集型实验之前，在大肠杆菌中完成表达筛选。重要的是，我们描述了一种基于极为成功的biGBac系统快速生成多顺反子细菌构建体的方法，这使得InteBac对从事重组蛋白复合物研究的研究人员特别感兴趣。