重组DNA
克隆(编程)
结扎
计算生物学
生物
遗传学
分子生物学
计算机科学
基因
程序设计语言
作者
Claire Strain-Damerell,P. Mahajan,Alejandra Fernández-Cid,O. Gileadi,N. Burgess-Brown
出处
期刊:Methods in molecular biology
日期:2020-10-31
卷期号:: 23-43
被引量:4
标识
DOI:10.1007/978-1-0716-0892-0_3
摘要
Structural genomics groups have identified the need to generate multiple truncated versions of each target to improve their success in producing a well-expressed, soluble, and stable protein and one that crystallizes and diffracts to a sufficient resolution for structural determination. At the Structural Genomics Consortium, we opted for the ligation-independent cloning (LIC) method which provides the throughput we desire to produce and screen many proteins in a parallel process. Here, we describe our LIC protocol for generating constructs in 96-well format and provide a choice of vectors suitable for expressing proteins in both E. coli and the baculovirus expression vector system (BEVS).
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