Synthetic BioBricks introduce novel capabilities to manipulate genetic information, direct transcription-translation processes, and program cellular behaviors in living organisms. To maintain the stability and functionality of synthetic BioBricks, assembled DNA fragments should be mutually compatible without inducing negative effects such as metabolic burden or cellular toxicity in host cells. However, a simple, rapid, and reliable method to evaluate BioBrick compatibility remains to be developed. In this study, we report BP (Blue/Purple, Ban/Pick) evaluation, a method utilizing chromoproteins to facilitate the identification of BioBrick compatibility in one-pot reactions. By visualizing and quantifying the ratio of blue to purple Escherichia coli (E. coli) colonies on LB-agar plates, we can easily validate the compatibility of desired BioBrick constructions. To demonstrate our design, we characterized BioBrick assemblies with antitoxin-toxin pair ccdA-ccdB, lysis protein E, or heterologous protein sfGFP. Among these, we successfully identified several compatible assemblies. We anticipate that BP evaluation will enhance biotechnological assessments of BioBrick compatibility in vivo and expand the application of chromoproteins in synthetic biology.

中文翻译：

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蓝紫色评估：色蛋白有助于鉴定 BioBrick 的相容性


合成 BioBricks 引入了操纵遗传信息、直接转录翻译过程和对生物体中的细胞行为进行编程的新功能。为了保持合成 BioBricks 的稳定性和功能，组装的 DNA 片段应相互兼容，而不会在宿主细胞中诱导代谢负荷或细胞毒性等负面影响。然而，一种简单、快速和可靠的方法来评估 BioBrick 兼容性仍有待开发。在本研究中，我们报告了 BP（蓝/紫、Ban/Pick）评估，这是一种利用色蛋白促进鉴定单锅反应中 BioBrick 相容性的方法。通过可视化和定量 LB 琼脂平板上蓝色与紫色大肠杆菌（大肠杆菌）菌落的比例，我们可以轻松验证所需 BioBrick 结构的兼容性。为了证明我们的设计，我们用抗毒素-毒素对 ccdA-ccdB、裂解蛋白 E 或异源蛋白 sfGFP 表征了 BioBrick 组装体。其中，我们成功确定了几个兼容的组件。我们预计 BP 评估将加强 BioBrick 体内相容性的生物技术评估，并扩大色蛋白在合成生物学中的应用。