Titlebook: Parasite Genomics Protocols; Sara E. Melville Book 2004 Humana Press 2004

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Transfection of the Human Malaria Parasite ,,placement, and transgene expression in this organism. These methods are detailed in this chapter, as are approaches used to monitor transfectants during the selection process. The different plasmid vectors that are currently used for gene targeting and transgene expression (including green fluoresce

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A PCR-Based Method for Gene Deletion and Protein Tagging in ,,. Here, we describe a polymerase chain reaction (PCR)-based method for direct gene deletion and the generation of epitope-tagged fusion proteins. The approach is based on methodologies developed for . and involves PCR amplification of a reporter cassette using primers containing flanking sequences s

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In Vitro Shuttle Mutagenesis Using Engineered Mariner Transposons,ch for gene identification and analysis is transposon mutagenesis. This can be performed directly in vivo, but often it is more convenient to generate transpositions in vitro for subsequent analysis in vivo, in a process termed “shuttle mutagenesis.” The . element . is well suited for application by

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Random Mutagenesis Strategies for Construction of Large and Diverse Clone Libraries of Mutated DNA Fragments,s strategy. This chapter describes three different methods for random mutagenesis, the use of XL1-red cells, error-prone polymerase chain reaction (PCR), and degenerate oligonucleotides-. (DOP). These mutagenesis strategies possess different benefits and pitfalls; thus, they are differentially usefu

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Separation, Digestion, and Cloning of Intact Parasite Chromosomes Embedded in Agarose,ntly at metaphase. Therefore, the development of pulsed field gel electrophoresis allowed the resolution of many parasite karyotypes for the first time. The ability to prepare intact chromosomes in agarose plugs and to isolate individual homologs by electrophoresis has led to many new applications i

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