用于鉴定条件必需基因的定义突变体文库测序（DML-Seq）
Defined Mutant Library Sequencing (DML-Seq) for Identification of Conditional Essential Genes.
作者信息
Shao Shuai, Wei Lifan, Xia Feng, Zhang Yuanxing, Wang And Qiyao
机构信息
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Department of Endodontics and Operative Dentistry, Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
出版信息
Bio Protoc. 2021 Mar 5;11(5):e3943. doi: 10.21769/BioProtoc.3943.
Transposon insertion sequencing (TIS) is an emerging technique which utilizes a massive transposon mutant library to screen specific phenotype and determine the conditional essential genetic requirements for bacterial fitness under distinct conditions combined with high-throughput parallel sequencing technology. Compared with a massive mutant library in traditional TIS, the defined mutant library sequencing (DML-Seq) has advantages as: 1) efficient mutagenesis; 2) low bottleneck effects; 3) avoid hotpots caused by screening; 4) can be directly used in the following experiments. Here, we described an optimized procedure of DML-Seq for fitness screen to supply classical TIS using the marine pathogenic bacterium as an example.
转座子插入测序（TIS）是一种新兴技术，它利用大规模转座子突变体文库来筛选特定表型，并结合高通量平行测序技术确定细菌在不同条件下适应度的条件必需基因要求。与传统TIS中的大规模突变体文库相比，定义突变体文库测序（DML-Seq）具有以下优点：1）高效诱变；2）低瓶颈效应；3）避免筛选导致的热点；4）可直接用于后续实验。在此，我们以海洋致病细菌为例，描述了一种优化的DML-Seq适应性筛选程序，以替代经典TIS。
Zotero 插件