检测mRNA开盖活力

摘要 mRNA decapping is a common step shared between two important mRNA decay pathways in yeast, Saccharomyces cerevisiae. To investigate how mRNAs are decapped, we have developed an assay that can be easily used to measure the decapping activity. This assay has been used to isolate yeast strains with altered decapping activities. The results demonstrated that decreased decapping activity in vitro corresponds well with the decapping-deÞcient phenotype in vivo. This assay has been applied to the puriÞed yeast decapping enzyme Dcp1 protein as well as crude yeast extracts and Xenopus oocyte extracts. 查看全文 Monitoring mRNA Decapping Activity   Shuang Zhang, Carol J. Williams,Michael Wormington,Audrey Stevens, and Stuart W. Peltz *Department of Molecular Genetics and Microbiology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey (UMDNJ), Piscataway, New Jersey 08854; †Graduate Program in Molecular Biosciences, Robert Wood Johnson Medical School, UMDNJ/Rutgers University, Piscataway,New Jersey 08854; \Cancer Institute of New Jersey, 675 Hoes Lane, Piscataway, New Jersey 08854;Department of Biology, University of Virginia, Charlottesville, Virginia 22903–2477; and Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831