Modular system for the construction of zinc-finger libraries and proteins
NATURE PROTOCOLS（2010）
摘要
Engineered zinc-finger transcription factors (ZF-TF) are powerful tools to modulate the expression of specific genes. Complex libraries of ZF-TF can be delivered into cells to scan the genome for genes responsible for a particular phenotype or to select the most effective ZF-TF to regulate an individual gene. In both cases, the construction of highly representative and unbiased libraries is critical. In this protocol, we describe a user-friendly ZF technology suitable for the creation of complex libraries and the construction of customized ZF-TFs. The new technology described here simplifies the building of ZF libraries, avoids PCR-introduced bias and ensures equal representation of every module. We also describe the construction of a customized ZF-TF that can be transferred to a number of expression vectors. This protocol can be completed in 9–11 d.
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关键词
cell biology,transfection,analytical chemistry,developmental biology,biophysics,microbiology,genetics,biochemistry,toxicology,rational drug design,electrophysiology,ecology,neuroscience,marine biology,escherichia coli,biomedical,zinc fingers,dna sequencing,proteomics,rna isolation,transduction,nature,biological chemistry,proteins,computational chemistry,endocrinology,antibodies,pharmacology,zinc finger,methodology,mutagenesis,physiology,transcription factors,nanotechnology,zoology,protein engineering,laboratory,bioinformatics,transcription factor,recombinant dna,recombinant proteins,forensics,parasitology,spectroscopy,computational biology,conservation biology,microarrays,plasmids,sequencing,plant biology,molecular biology,protocols,cell signalling,immunology,gene expression,glycomics,systems biology,genomics,biotechnology,microscopy,science and technology,drug discovery,protein synthesis,structural biology,chemistry,evolutionary biology,histology
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