Sola-Landa Alberto, Rodríguez-García Antonio, Barreiro Carlos, Pérez-Redondo Rosario
Instituto de Biotecnología de León (INBIOTEC), Parque Científico de León, Avenida Real 1, 24006, León, Spain.
Área de Microbiología, Departamento de Biología Molecular, Campus de Ponferrada, Universidad de León, León, Spain.
Methods Mol Biol. 2017;1645:45-58. doi: 10.1007/978-1-4939-7183-1_4.
The Next-Generation Sequencing technology has enormously eased the bacterial genome sequencing and several tens of thousands of genomes have been sequenced during the last 10 years. Most of the genome projects are published as draft version, however, for certain applications the complete genome sequence is required.In this chapter, we describe the strategy that allowed the complete genome sequencing of Mycobacterium neoaurum NRRL B-3805, an industrial strain exploited for steroid production, using Ion Torrent sequencing reads and the genome of a close strain as the reference. This protocol can be applied to analyze the genetic variations between closely related strains; for example, to elucidate the point mutations between a parental strain and a random mutagenesis-derived mutant.
新一代测序技术极大地简化了细菌基因组测序，在过去十年中已有数以万计的基因组被测序。然而，大多数基因组计划都是以草图版本发布的，不过对于某些应用而言，完整的基因组序列是必需的。在本章中，我们描述了一种策略，该策略利用离子激流测序读数，并以一个近缘菌株的基因组作为参考，实现了新金色分枝杆菌NRRL B - 3805（一种用于甾体生产的工业菌株）的全基因组测序。该方案可用于分析亲缘关系密切的菌株之间的遗传变异；例如，阐明亲本菌株与随机诱变衍生突变体之间的点突变。
