Springer Protocols: Abstract: Localization of Proteins and Organelles Using Fluorescence Microscopy

Contents of this article

Browse by Subject

Upload a Protocol

Upload your own protocols for personal use.

Protocol Alert

Receive e-mail notifications about new content on Springer Protocols.

Video Protocols

View our video protocols.

Call for video protocols.

Comments

Read comments by other users and add your own.

Favorites

Save your favorite protocols to your My Protocols area.

RSS

RSS Feeds

Inside Springer Protocols

Useful Tools

Localization of Proteins and Organelles Using Fluorescence Microscopy

By: Jean-Claude Farre², Kanae Shirahama-Noda², Lan Zhang², Keith Booher², Suresh Subramani²

Abstract

Full Text

Download PDF (301K)

This chapter describes the different methods used for localization of proteins and organelles in Pichia pastoris. A series of plasmids and a modified immunofluorescence protocol for localization and co-localization of proteins and organelles are described. Also included are protocols for the labeling of different subcellular organelles with vital stains.

Affiliation(s): (2) Section of Molecular Biology, Division of Biological Sciences, University of California San Diego, La Jolla, CA

Book Title: Pichia Protocols

Series: Methods in Molecular Biology | Volume: 389 | Pub. Date: Aug-08-2007 | Page Range: 239-249 | DOI: 10.1007/978-1-59745-456-8_17

Subject: Biotechnology

Key Words: Protein localization - protein colocalization - fluorescence microscopy - organelle markers - green fluorescent protein variants - peroxisomes

References

Download References

1.	Miyawaki, A., Llopis, J., Heim, R., et al. (1997) Fluorescent indicators for Ca²⁺ based on green fluorescent proteins and calmodulin. Nature 388, 882–887.

2.	Rossanese, O. W., Soderholm, J., Bevis, B. J., et al. (1999) Golgi structure correlates with transitional endoplasmic reticulum organization in Pichia pastoris and Saccharomyces cerevisiae. J. Cell Biol. 145, 69–81.

3.	Vida, T. A. and Emr, S. D. (1995) A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J. Cell Biol. 128, 779–792.

4.	Dann, O., Bergen, G., Demant, E., and Voltz, G. (1971) Trypanocide Diamidine des 2-Phenyl-benofurans, 2-Phenyl-indens und 2-Phenyl-indols. Liebigs Ann. Chemi. 749, 68–89.

5.	Gould, S. J., Keller, G. A., and Subramani, S. (1987) Identification of a peroxisomal targeting signal at the carboxy terminus of firefly luciferase. J. Cell Biol. 105, 2923–2931.

6.	Campbell, R. E., Tour, O., Palmer, A. E., et al. (2002) A monomeric red fluorescent protein. Proc. Natl. Acad. Sci. USA 99, 7877–7882.

7.	Wiemer, E. A., Luers, G. H., Faber, K. N., Wenzel, T., Veenhuis, M., and Subramani, S. (1996) Isolation and characterization of Pas2p, a peroxisomal membrane protein essential for peroxisome biogenesis in the methylotrophic yeast Pichia pastoris. J. Biol. Chem. 271, 18,973–18,980.

8.	Sears, I. B., O’Connor, J., Rossanese, O. W., and Glick, B. S. (1998) A versatile set of vectors for constitutive and regulated gene expression in Pichia pastoris. Yeast 14, 783–790.

Comments (Loading...)

Post comment/View all comments