Literature DB >> 34079146

Implementing a Hybrid Expression Method That Allows Upper-Division Biochemistry Lab Students To Engage in a Full Protein Production Experience While Allowing Ample Time for Characterization Experiments.

Josiah W Johnson1, Christian D Mitchell1, Anna M Deloach1, Hannah E Simpson1, Tori B Dunlap1.   

Abstract

Protein structure, function, and signaling are a large portion of biochemistry. Because of this, proteins are often used as model systems in biochemistry laboratory courses, where a course-long project might comprise protein expression, purification, and characterization. Two common protein expression methods are isopropyl β-d-1-thiogalactopyranoside (IPTG) induction, which utilizes easy-to-make media but requires extensive cell-growth monitoring that is time-intensive, and autoinduction, which employs multicomponent media that are time-consuming to make but require no cell-growth monitoring. A protein expression method that is a hybrid of IPTG induction and autoinduction is presented. The hybrid method utilizes the medium of IPTG induction and the no-cell-growth-monitoring induction process of autoinduction, saving hands-on time in the protein expression phase to allow more time for protein characterization while still having students execute each step.

Entities:  

Keywords:  Biochemistry; Enzymes; Graduate Education/Research; Hands-On Learning/Manipulatives; Laboratory Instruction; Proteins/Peptides; Upper-Division Undergraduate

Year:  2019        PMID: 34079146      PMCID: PMC8168722          DOI: 10.1021/acs.jchemed.8b00674

Source DB:  PubMed          Journal:  J Chem Educ        ISSN: 0021-9584            Impact factor:   2.979


  20 in total

1.  A new role for IQ motif proteins in regulating calmodulin function.

Authors:  John A Putkey; Quinn Kleerekoper; Tara R Gaertner; M Neal Waxham
Journal:  J Biol Chem       Date:  2003-10-09       Impact factor: 5.157

2.  Evidencing the role of lactose permease in IPTG uptake by Escherichia coli in fed-batch high cell density cultures.

Authors:  Alfred Fernández-Castané; Claire E Vine; Glòria Caminal; Josep López-Santín
Journal:  J Biotechnol       Date:  2011-12-21       Impact factor: 3.307

Review 3.  Protein production and purification.

Authors:  Susanne Gräslund; Pär Nordlund; Johan Weigelt; B Martin Hallberg; James Bray; Opher Gileadi; Stefan Knapp; Udo Oppermann; Cheryl Arrowsmith; Raymond Hui; Jinrong Ming; Sirano dhe-Paganon; Hee-won Park; Alexei Savchenko; Adelinda Yee; Aled Edwards; Renaud Vincentelli; Christian Cambillau; Rosalind Kim; Sung-Hou Kim; Zihe Rao; Yunyu Shi; Thomas C Terwilliger; Chang-Yub Kim; Li-Wei Hung; Geoffrey S Waldo; Yoav Peleg; Shira Albeck; Tamar Unger; Orly Dym; Jaime Prilusky; Joel L Sussman; Ray C Stevens; Scott A Lesley; Ian A Wilson; Andrzej Joachimiak; Frank Collart; Irina Dementieva; Mark I Donnelly; William H Eschenfeldt; Youngchang Kim; Lucy Stols; Ruying Wu; Min Zhou; Stephen K Burley; J Spencer Emtage; J Michael Sauder; Devon Thompson; Kevin Bain; John Luz; Tarun Gheyi; Fred Zhang; Shane Atwell; Steven C Almo; Jeffrey B Bonanno; Andras Fiser; Sivasubramanian Swaminathan; F William Studier; Mark R Chance; Andrej Sali; Thomas B Acton; Rong Xiao; Li Zhao; Li Chung Ma; John F Hunt; Liang Tong; Kellie Cunningham; Masayori Inouye; Stephen Anderson; Heleema Janjua; Ritu Shastry; Chi Kent Ho; Dongyan Wang; Huang Wang; Mei Jiang; Gaetano T Montelione; David I Stuart; Raymond J Owens; Susan Daenke; Anja Schütz; Udo Heinemann; Shigeyuki Yokoyama; Konrad Büssow; Kristin C Gunsalus
Journal:  Nat Methods       Date:  2008-02       Impact factor: 28.547

4.  Purification of cyclic 3',5'-nucleotide phosphodiesterase inhibitory protein by affinity chromatography on activator protein coupled to Sepharose.

Authors:  C B Klee; M H Krinks
Journal:  Biochemistry       Date:  1978-01-10       Impact factor: 3.162

5.  Affinity chromatography purification of a cyclic nucleotide phosphodiesterase using immobilized modulator protein, a troponin C-like protein from brain.

Authors:  D M Watterson; T C Vanaman
Journal:  Biochem Biophys Res Commun       Date:  1976-11-08       Impact factor: 3.575

Review 6.  Regulation of the calmodulin-stimulated protein phosphatase, calcineurin.

Authors:  C B Klee; H Ren; X Wang
Journal:  J Biol Chem       Date:  1998-05-29       Impact factor: 5.157

7.  Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems.

Authors:  Zhaopeng Li; Wolfgang Kessler; Joop van den Heuvel; Ursula Rinas
Journal:  Appl Microbiol Biotechnol       Date:  2011-06-23       Impact factor: 4.813

8.  Calmodulin target database.

Authors:  K L Yap; J Kim; K Truong; M Sherman; T Yuan; M Ikura
Journal:  J Struct Funct Genomics       Date:  2000

9.  Purification and characterization of enzymes from yeast: an extended undergraduate laboratory sequence for large classes.

Authors:  Kelly E Johanson; Terry J Watt; Neil R McIntyre; Marleesa Thompson
Journal:  Biochem Mol Biol Educ       Date:  2013 Jul-Aug       Impact factor: 1.160

Review 10.  High-throughput recombinant protein expression in Escherichia coli: current status and future perspectives.

Authors:  Baolei Jia; Che Ok Jeon
Journal:  Open Biol       Date:  2016-08       Impact factor: 6.411

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