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Transposon Mutagenesis of Rhodobacter sphaeroides

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Author:
,
Subject:
Science and Technology
Institution Name:
University of Wisconsin-Madison
Collection:
Association for Biology Laboratory Education (ABLE)
Grade Level:
Post-secondary
Abstract:

Creation and characterization of mutants is the basis for any genetic analysis. This exercise demonstrates a simple, safe procedure for transposon mutagenesis of Rhodobacter sphaeroides, a purple non-sulfur photosynthetic bacteria. Students perform the mutagenesis by mating a transposon-carrying plasmid from Escherichia coli to R. sphaeroides and then selecting for the drug resistance carried on the transposon. Only R. sphaeroides carrying the transposon in the chromosome survive the selection. Transposon carrying mutants are then scored for various phenotypes. The metabolic diversity of R. sphaeroides allows the isolation of nutritional, photopigment, and photosynthetic mutants. Further analysis of mutants is possible.

Course Type:
Learning Module
Languages:
English
Material Type:
Activities and Labs, Lesson Plans
Media Format:
Downloadable docs
Conditions of Use:
Custom Permissions
All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without the prior written permission of the copyright owner. Use solely at one's own institution with no intent for profit is excluded from the preceding copyright restriction, unless otherwise noted on the copyright notice of the individual chapter in this volume. Proper credit to this publication must be included in your laboratory outline for each use; a sample citation is given [on the copyright page of each volume]. Upon obtaining permission or with the "sole use at one's own institution" exclusion, ABLE strongly encourages individuals to use the exercises in this proceedings volume in their teaching program.

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