McKeown, Lynn, Jones, Vicky Claire and Jones, Owen T. (2009) PIN-G reporter for imaging and defining trafficking signals in membrane proteins. In: Bioluminescence. Methods in Molecular Biology (574). Humana Press, New York, NY, USA, pp. 235-248. ISBN 978-1-60327-320-6
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Official URL: http://dx.doi.org/10.1007/978-1-60327-321-3_19
The identification of motifs that control the intracellular trafficking of proteins is a fundamental objective of cell biology. Once identified, such regions should, in principle, be both necessary and sufficient to direct any randomly distributed protein, acting as a reporter, to the subcellular compartment in question. However, most reporter proteins have limited versatility owing to their endogenous expression and limited modes of detection--especially in live cells. To surmount such limitations, we engineered a plasmid--pIN-G--encoding an entirely artificial, type I transmembrane reporter protein (PIN-G), containing HA, cMyc and GFP epitope, and fluorescence tags. Although originally designed for trafficking studies, pIN technology is a powerful tool applicable to almost every area of biology. Here we describe the methodologies used routinely in analyzing pIN constructs and some of their derivatives.
|Item Type:||Book Section|
|Subjects:||C - Biological sciences > C100 - Biology|
|Schools:||Faculty of Clinical & Biomedical Sciences > School of Pharmacy and Biomedical Sciences|
|Deposited By:||Vicky Claire Jones|
|Deposited On:||06 Jun 2013 13:37|
|Last Modified:||17 May 2016 12:42|
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