Blue-Purple evaluation: Chromoproteins facilitate the identification of BioBrick compatibility

IF 3.5 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology and Bioengineering Pub Date : 2024-10-14 DOI:10.1002/bit.28862

Fang Ba, Yufei Zhang, Luyao Wang, Wan-Qiu Liu, Jian Li

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引用次数: 0

Abstract

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.

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蓝紫色评价：色素蛋白有助于鉴定生物特异性

合成生物砖（Synthetic BioBricks）具有操纵遗传信息、指导转录-翻译过程以及对生物体内的细胞行为进行编程的新功能。为保持合成生物砖的稳定性和功能性，组装的 DNA 片段应相互兼容，而不会对宿主细胞造成代谢负担或细胞毒性等负面影响。然而，评估 BioBrick 兼容性的简单、快速、可靠的方法仍有待开发。在这项研究中，我们报告了 BP（蓝/紫，Ban/Pick）评估方法，这是一种利用色素蛋白促进在一锅反应中鉴定 BioBrick 兼容性的方法。通过可视化和量化 LB-agar 平板上蓝色和紫色大肠杆菌（E. coli）菌落的比例，我们可以轻松验证所需 BioBrick 构建的兼容性。为了证明我们的设计，我们对带有抗毒素-毒素对 ccdA-ccdB、裂解蛋白 E 或异源蛋白 sfGFP 的 BioBrick 组装进行了鉴定。其中，我们成功鉴定了几个兼容的组装体。我们预计 BP 评估将加强生物技术评估 BioBrick 在体内的兼容性，并扩大色蛋白在合成生物学中的应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology and Bioengineering 工程技术-生物工程与应用微生物

CiteScore

7.90

自引率

5.30%

发文量

280

审稿时长

2.1 months

期刊介绍： Biotechnology & Bioengineering publishes Perspectives, Articles, Reviews, Mini-Reviews, and Communications to the Editor that embrace all aspects of biotechnology. These include: -Enzyme systems and their applications, including enzyme reactors, purification, and applied aspects of protein engineering -Animal-cell biotechnology, including media development -Applied aspects of cellular physiology, metabolism, and energetics -Biocatalysis and applied enzymology, including enzyme reactors, protein engineering, and nanobiotechnology -Biothermodynamics -Biofuels, including biomass and renewable resource engineering -Biomaterials, including delivery systems and materials for tissue engineering -Bioprocess engineering, including kinetics and modeling of biological systems, transport phenomena in bioreactors, bioreactor design, monitoring, and control -Biosensors and instrumentation -Computational and systems biology, including bioinformatics and genomic/proteomic studies -Environmental biotechnology, including biofilms, algal systems, and bioremediation -Metabolic and cellular engineering -Plant-cell biotechnology -Spectroscopic and other analytical techniques for biotechnological applications -Synthetic biology -Tissue engineering, stem-cell bioengineering, regenerative medicine, gene therapy and delivery systems The editors will consider papers for publication based on novelty, their immediate or future impact on biotechnological processes, and their contribution to the advancement of biochemical engineering science. Submission of papers dealing with routine aspects of bioprocessing, description of established equipment, and routine applications of established methodologies (e.g., control strategies, modeling, experimental methods) is discouraged. Theoretical papers will be judged based on the novelty of the approach and their potential impact, or on their novel capability to predict and elucidate experimental observations.