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Genetic analysis of the RNA polymerase II CTD in Drosophila.

Methods. 2019; 
LuFeiyue,GilmourDav
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PCR Cloning and Subcloning … Fig. 4. Testing the functionality of UAS-Rpb1 derivatives using the Rpb1 G0040 allele … from genomic DNA or subclones of the amplified DNA, although we occasionally order 200 bp to 1 kb synthetic DNA fragments (Integrated DNA Technologies or Genscript) for sequences that … Get A Quote

摘要

The Carboxy-terminal Domain (CTD) of RNA polymerase II (Pol II) plays essential roles in regulating gene expression in eukaryotes. Here, we describe multiple genetic approaches for studying the CTD in Drosophila that complement pre-existing molecular analyses of the Pol II CTD in other experimental models. These approaches will allow one to assess the effects of any CTD mutations in a developmentally complex organism. The approaches discussed in this work can in principle, be applied to analyze other transcription components in eukaryotes.

关键词

CRISPR/Cas9,Drosophila,GAL4/UAS,Genetics,Pol II