GROQ-seq Function Measurements for TEV Protease SSVL and epPCR Libraries.

This dataset contains GROQ-seq functional measurements of TEV protease variant libraries — a site-saturation variant library (SSVL) and an error-prone PCR (epPCR) library — covering over 18,000 protease variants across 24 experimental conditions. Fitness is measured via a split-DHFR system in which the TEV protease substrate is embedded as the linker of a split dihydrofolate reductase: TEV cleavage destroys DHFR and reduces trimethoprim resistance, translating protease activity into differences in cellular growth. Conditions vary independently tuned levels of TEV expression, split-DHFR expression, and trimethoprim selection. This run of the assay was performed at the Living Measurement Systems Foundry (LMSF) at the National Institute of Standards and Technology. Part of a broader effort to generate sequence → function data for proteases, enabling predictive models of protease activity and specificity with applications in protein engineering and therapeutic development.本数据集包含烟草蚀刻病毒（TEV）蛋白酶变体库的GROQ-seq功能检测数据，所涉变体库包括定点饱和突变变体库（site-saturation variant library，SSVL）与易错PCR（error-prone PCR，epPCR）文库，覆盖24种实验条件下的18000余种蛋白酶变体。变体的功能适配度通过分裂二氢叶酸还原酶（split-DHFR）系统测定：该系统将TEV蛋白酶的识别底物作为连接肽段嵌入分裂二氢叶酸还原酶中，TEV蛋白酶的切割会破坏分裂二氢叶酸还原酶的结构完整性，降低菌体对甲氧苄啶（trimethoprim）的抗性，进而将蛋白酶活性转化为细胞生长水平的差异。各实验条件分别独立调控TEV蛋白酶表达量、分裂二氢叶酸还原酶表达量以及甲氧苄啶筛选压力。 本次检测实验于美国国家标准与技术研究院（National Institute of Standards and Technology，NIST）的活体测量系统制备工坊（Living Measurement Systems Foundry，LMSF）完成。本数据集属于构建蛋白酶序列-功能数据体系的整体研究的一部分，旨在构建蛋白酶活性与特异性的预测模型，相关成果可应用于蛋白质工程及治疗药物开发领域。