Tandem Affinity Purification in Drosophila Heads and Ovaries

Tandem affinity purification in Drosophila: the advantages of the GS-TAP system.

Tandem affinity purification (TAP) has been widely used for the analysis of protein complexes. We investigated the parameters of the recently developed TAP method (GS-TAP) and its application in Drosophila. This new tag combination includes two Protein G modules and a streptavidin binding peptide (SBP), separated by one or two TEV protease cleavage sites. We made pMK33-based GS-TAP vectors to a...

Purification of Transcripts and Metabolites from Drosophila Heads

For the last decade, we have tried to understand the molecular and cellular mechanisms of neuronal degeneration using Drosophila as a model organism. Although fruit flies provide obvious experimental advantages, research on neurodegenerative diseases has mostly relied on traditional techniques, including genetic interaction, histology, immunofluorescence, and protein biochemistry. These techniq...

Application and Prospects of Tandem Affinity Purification Method

Tandem affinity purification of miRNA target mRNAs (TAP-Tar)

MicroRNAs (miRNAs) bind to Argonaute proteins, and together they form the RISC complex and regulate target mRNA translation and/or stability. Identification of mRNA targets is key to deciphering the physiological functions and mode of action of miRNAs. In mammals, miRNAs are generally poorly homologous to their target sequence, and target identification cannot be based solely on bioinformatics....

Tandem Affinity Purification to Enhance Interacting Protein Identification

With the number of complete genome sequences increasing at an ever-faster pace, we are getting sequence data for the full complement of proteins or “proteome” (Wasinger et al. 1995; Wilkins et al. 1996), encoded by various organisms. To understand how the proteome confers specific properties to a host cell, one needs to obtain information regarding several parameters, including the level of exp...