Prev. Article Next Article Table of Contents

Article

Identification of Yeast V-ATPase Mutants by Western Blots Analysis of Whole Cell Lysates

Your current credentials do not allow retrieval of the full text.

Purchase the full-text

PDF/HTML,
figures/images,
references and tables,
(where available)

Karlett Parra-Belky

Department of Chemistry, Ball State University, Muncie, IN 47306

J. Chem. Educ., 2002, 79 (11), p 1348

DOI: 10.1021/ed079p1348

Publication Date (Web): November 1, 2002

Section:

History, Education, and Documentation

Abstract

A biochemistry laboratory was designed for an undergraduate course to help students better understand the link between molecular engineering and biochemistry. Students identified unknown yeast strains with high specificity using SDS-PAGE and Western blot analysis of whole cell lysates. This problem-solving exercise is a common application of biochemistry in biotechnology research. Three different strains were used: a wild-type and two mutants for the proton pump vacuolar ATPase (V-ATPase). V-ATPases are multisubunit enzymes and the mutants used were deletion mutants; each lacked one structural gene of the complex. After three, three-hour labs, mutant strains were easily identified by the students and distinguished from wild-type cells analyzing the pattern of SDS-PAGE distribution of proteins. Identifying different subunits of one multimeric protein allowed for discussion of the structure and function of this metabolic enzyme, which captured the interest of the students. The experiment can be adapted to other multimeric protein complexes and shows improvement of the described methodology over previous reports, perhaps because the problem and its solution are representative of the type of techniques currently used in research labs.

Keywords (Audience):

Upper-Division Undergraduate

Keywords (Domain):

Biochemistry

Keywords (Pedagogy):

Hands-On Learning / Manipulatives

Keywords (Subject):

Biotechnology

View: Hi-Res PDF | PDF w/ Links

Citing Articles

Citation data is made available by participants in CrossRef's Cited-by Linking service. For a more comprehensive list of citations to this article, users are encouraged to perform a search in SciFinder.

This article has been cited by 3 ACS Journal articles (3 most recent appear below).

Determination of the Subunit Molecular Mass and Composition of Alcohol Dehydrogenase by SDS-PAGE
Barbara T. Nash
Journal of Chemical Education2007 84 (9), 1508
- Determination of the Subunit Molecular Mass and Composition of Alcohol Dehydrogenase by SDS-PAGE
  Barbara T. Nash
  Journal of Chemical Education2007 84 (9), 1508
  SDS-PAGE is a simple, rapid technique that has many uses in biochemistry and is readily adaptable to the undergraduate laboratory. It is, however, a technique prone to several types of procedural pitfalls. This article describes the use of SDS-PAGE to ...
  Abstract | Hi-Res PDF | PDF w/ Links
Preparation, Purification, and Secondary Structure Determination of Bacillus circulans Xylanase. A Modular Laboratory Incorporating Aspects of Molecular Biology, Biochemistry, and Biophysical Chemistry
Sal Russo and Lisa Gentile
Journal of Chemical Education2006 83 (12), 1850
- Preparation, Purification, and Secondary Structure Determination of Bacillus circulans Xylanase. A Modular Laboratory Incorporating Aspects of Molecular Biology, Biochemistry, and Biophysical Chemistry
  Sal Russo and Lisa Gentile
  Journal of Chemical Education2006 83 (12), 1850
  An integrated six-week module that allows students to develop biochemistry laboratory techniques in the context of a discovery-oriented project has been developed. This module, suitable for undergraduate third- or fourth-year biochemistry lab, focuses on ...
  Abstract | Hi-Res PDF | PDF w/ Links
An SDS-PAGE Examination of Protein Quaternary Structure and Disulfide Bonding for a Biochemistry Laboratory
Jennifer L. Powers , Carla S. Andrews , Caroline C. St. Antoine , Swapan S. Jain and Vicky L. H. Bevilacqua
Journal of Chemical Education2005 82 (1), 93
- An SDS-PAGE Examination of Protein Quaternary Structure and Disulfide Bonding for a Biochemistry Laboratory
  Jennifer L. Powers , Carla S. Andrews , Caroline C. St. Antoine , Swapan S. Jain and Vicky L. H. Bevilacqua
  Journal of Chemical Education2005 82 (1), 93
  Electrophoresis is a valuable tool for biochemists, yet this technique is often not included in biochemistry laboratory curricula owing to time constraints or lack of equipment. Protein structure is also a topic of interest in many disciplines, yet most ...
  Abstract | Hi-Res PDF | PDF w/ Links