Ortega Claudia, Abreu Cecilia, Oppezzo Pablo, Correa Agustín
Recombinant Protein Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay.
Research Laboratory on Chronic Lymphocytic Leukemia, Institut Pasteur de Montevideo, Montevideo, Uruguay.
Methods Mol Biol. 2019;2025:3-32. doi: 10.1007/978-1-4939-9624-7_1.
The advent of new DNA sequencing technologies leads to a dramatic increase in the number of available genome sequences and therefore of target genes with potential for functional analysis. The insertion of these sequences into proper expression vectors requires a simple an efficient cloning method. In addition, when expressing a target protein, quite often it is necessary to evaluate different DNA constructs to achieve a soluble and homogeneous expression of the target with satisfactory yields. The development of new molecular methods made possible the cloning of a huge number of DNA sequences in a high-throughput manner, necessary for meeting the increasing demands for soluble protein expression and characterization. In this chapter several molecular methods suitable for high-throughput cloning are reviewed.
新DNA测序技术的出现使得可用基因组序列的数量大幅增加,从而使具有功能分析潜力的靶基因数量也大幅增加。将这些序列插入合适的表达载体需要一种简单而高效的克隆方法。此外,在表达靶蛋白时,通常有必要评估不同的DNA构建体,以实现靶标的可溶性和均一性表达,并获得令人满意的产量。新分子方法的发展使以高通量方式克隆大量DNA序列成为可能,这对于满足对可溶性蛋白表达和表征日益增长的需求是必要的。本章将综述几种适用于高通量克隆的分子方法。