Structural and biophysical characterization of molecular mechanisms of disease‐causing pathogens, such as Mycobacterium tuberculosis, often requires recombinant expression of large amounts highly pure protein. For the production of mycobacterial proteins, overexpression in the fast‐growing and non‐pathogenic species Mycobacterium smegmatis has several benefits over the standard Escherichia coli expression strains. However, unlike for E. coli, the range of expression vectors currently available is limited. Here we describe the development of the pMy vector series, a set of expression plasmids for recombinant production of single proteins and protein complexes in M. smegmatis. By incorporating an alternative selection marker, we show that these plasmids can also be used for co‐expression studies. All vectors in the pMy vector series are available in the Addgene repository (www.addgene.com).

中文翻译：

---

pMy载体系列：在耻垢分枝杆菌中重组生产分枝杆菌蛋白的多功能克隆平台

致病性病原体（例如结核分枝杆菌）的分子机制的结构和生物物理表征通常需要重组表达大量高纯度蛋白质。对于分枝杆菌蛋白质的生产，在快速增长且非致病性物种中，耻垢分枝杆菌比标准大肠杆菌表达菌株具有多个优势。但是，与大肠杆菌不同，目前可用的表达载体的范围是有限的。在这里，我们描述了pMy载体系列的开发，pMy载体系列是一套用于重组生产耻垢分枝杆菌中的单个蛋白质和蛋白质复合物的表达质粒。通过掺入替代选择标记，我们表明这些质粒也可用于共表达研究。pMy载体系列中的所有载体均可在Addgene存储库（www.addgene.com）中找到。