The Resource Membrane proteins-- production and functional characterization, edited by Arun K. Shukla

Membrane proteins-- production and functional characterization, edited by Arun K. Shukla

Label
Membrane proteins-- production and functional characterization
Title
Membrane proteins-- production and functional characterization
Statement of responsibility
edited by Arun K. Shukla
Contributor
Editor
Subject
Genre
Language
  • eng
  • eng
Member of
Cataloging source
MiAaPQ
Dewey number
572.696
Illustrations
  • illustrations
  • photographs
  • plates
Index
index present
Language note
English
LC call number
QP552.M44
LC item number
.M463 2015
Literary form
non fiction
Nature of contents
  • dictionaries
  • bibliography
http://library.link/vocab/relatedWorkOrContributorName
Shukla, Arun K.
Series statement
Methods in Enzymology
Series volume
Volume 556
http://library.link/vocab/subjectName
Membrane proteins
Label
Membrane proteins-- production and functional characterization, edited by Arun K. Shukla
Instantiates
Publication
Copyright
Note
Description based upon print version of record
Bibliography note
Includes bibliographical references at the end of each chapters and indexes
Carrier category
online resource
Carrier category code
cr
Content category
text
Content type code
txt
Contents
  • Front Cover; Membrane Proteins-Production and Functional Characterization; Copyright; Contents; Contributors; Preface; Section I: Recombinant Expression of Membrane Proteins; Chapter 1: Engineering Escherichia coli for Functional Expression of Membrane Proteins; 1. Introduction; 1.1. Green fluorescent protein as a folding reporter; 1.2. Antibiotic resistance marker protein for selection; 2. Preparation of Erythromycin-Sensitive E. coli Strain; 3. Preparation of Expression Plasmid; 3.1. Construction of expression vector for target protein-GFP-ErmC fusion; 3.2. Expression test
  • 3.2.1. Bacterial culture for protein expression and determination of whole-cell fluorescence3.2.2. Gel-based analysis of the expressed fusion constructs; 4. Selecting Cells for Better Expression; 5. Characterizing Evolved Strains; 5.1. Basic characterizations; 5.1.1. Plasmid copy number, DNA sequencing, and transcript levels; 5.2. Plasmid curing; 5.3. Functional assays; 6. Summary; Acknowledgments; References; Chapter 2: ACEMBLing a Multiprotein Transmembrane Complex: The Functional SecYEG-SecDF-YajC-YidC Holotranslocon Protein S...; 1. Introduction; 2. ACEMBLing the HTL Multiprotein Complex
  • 3. Purifying the HTL4. HTL Integrity and Activity; 4.1. Incorporation of translocation complexes in proteoliposomes; 4.2. Orientation of the reconstituted complexes; 4.3. Subunit interactions and activity of reconstituted translocation complexes; 4.3.1. ATP-stimulated protein secretion by SecA ATPase; 4.3.2. PMF-stimulated protein secretion activity; 4.3.3. Membrane protein insertion activity of the HTL; 5. Discussion and Conclusions; Acknowledgments; References; Chapter 3: Expression and Purification of OsVDAC4; 1. Introduction; 2. Equipment; 3. Materials; 3.1. Solutions and buffers
  • 4. Protocols4.1. Cloning; 4.2. Transformation protocol for protein expression; Duration: 2 h + overnight incubation; 4.3. Screening for colonies with high protein expression; Tips; 4.4. Expression and purification of OsVDAC4; 4.4.1. Soluble OsVDAC4 protein purification; Duration: 2-3 days; Tips; 4.4.2. OsVDAC4 purification from inclusion bodies; Duration: 2 days; Tips; 4.5. Tryptic digestion of OsVDAC4 for mass spectrometry; Duration: 2-3 days; 4.6. Functional characterization of OsVDAC4; 4.6.1. Liposome swelling assay; Duration: 2 days; Tips; Note; 4.6.2. Planar bilayer lipid membrane
  • Duration: 5-6hTips; Note; Acknowledgments; References; Chapter 4: Membrane Protein Expression in Lactococcus lactis; 1. Introduction; 2. Equipment and Materials; 3. Buffers and Media; 4. Protocol; 4.1. Preparation; 4.2. Duration; 4.3. Tip; 5. Step 1: Cloning the Target Gene into pNZ8048 and Transformation into L. lactis; 5.1. Overview; Amplification of the Target Gene by PCR; Restriction digestion; Preparation of pNZ vector; Restriction digestion; Ligation and pellet paint coprecipitation; Preparation of electrocompetent L. lactis; Transformation into electrocompetent L. lactis cells
  • 5.2. Tips
Dimensions
unknown
Edition
First edition.
Extent
1 online resource (693 p.)
Form of item
online
Isbn
9780128015216
Media category
computer
Media type code
c
Specific material designation
remote
System control number
  • (CKB)3710000000385534
  • (EBL)2009776
  • (SSID)ssj0001534340
  • (PQKBManifestationID)12551762
  • (PQKBTitleCode)TC0001534340
  • (PQKBWorkID)11494957
  • (PQKB)11088913
  • (MiAaPQ)EBC2009776
  • (EXLCZ)993710000000385534
Label
Membrane proteins-- production and functional characterization, edited by Arun K. Shukla
Publication
Copyright
Note
Description based upon print version of record
Bibliography note
Includes bibliographical references at the end of each chapters and indexes
Carrier category
online resource
Carrier category code
cr
Content category
text
Content type code
txt
Contents
  • Front Cover; Membrane Proteins-Production and Functional Characterization; Copyright; Contents; Contributors; Preface; Section I: Recombinant Expression of Membrane Proteins; Chapter 1: Engineering Escherichia coli for Functional Expression of Membrane Proteins; 1. Introduction; 1.1. Green fluorescent protein as a folding reporter; 1.2. Antibiotic resistance marker protein for selection; 2. Preparation of Erythromycin-Sensitive E. coli Strain; 3. Preparation of Expression Plasmid; 3.1. Construction of expression vector for target protein-GFP-ErmC fusion; 3.2. Expression test
  • 3.2.1. Bacterial culture for protein expression and determination of whole-cell fluorescence3.2.2. Gel-based analysis of the expressed fusion constructs; 4. Selecting Cells for Better Expression; 5. Characterizing Evolved Strains; 5.1. Basic characterizations; 5.1.1. Plasmid copy number, DNA sequencing, and transcript levels; 5.2. Plasmid curing; 5.3. Functional assays; 6. Summary; Acknowledgments; References; Chapter 2: ACEMBLing a Multiprotein Transmembrane Complex: The Functional SecYEG-SecDF-YajC-YidC Holotranslocon Protein S...; 1. Introduction; 2. ACEMBLing the HTL Multiprotein Complex
  • 3. Purifying the HTL4. HTL Integrity and Activity; 4.1. Incorporation of translocation complexes in proteoliposomes; 4.2. Orientation of the reconstituted complexes; 4.3. Subunit interactions and activity of reconstituted translocation complexes; 4.3.1. ATP-stimulated protein secretion by SecA ATPase; 4.3.2. PMF-stimulated protein secretion activity; 4.3.3. Membrane protein insertion activity of the HTL; 5. Discussion and Conclusions; Acknowledgments; References; Chapter 3: Expression and Purification of OsVDAC4; 1. Introduction; 2. Equipment; 3. Materials; 3.1. Solutions and buffers
  • 4. Protocols4.1. Cloning; 4.2. Transformation protocol for protein expression; Duration: 2 h + overnight incubation; 4.3. Screening for colonies with high protein expression; Tips; 4.4. Expression and purification of OsVDAC4; 4.4.1. Soluble OsVDAC4 protein purification; Duration: 2-3 days; Tips; 4.4.2. OsVDAC4 purification from inclusion bodies; Duration: 2 days; Tips; 4.5. Tryptic digestion of OsVDAC4 for mass spectrometry; Duration: 2-3 days; 4.6. Functional characterization of OsVDAC4; 4.6.1. Liposome swelling assay; Duration: 2 days; Tips; Note; 4.6.2. Planar bilayer lipid membrane
  • Duration: 5-6hTips; Note; Acknowledgments; References; Chapter 4: Membrane Protein Expression in Lactococcus lactis; 1. Introduction; 2. Equipment and Materials; 3. Buffers and Media; 4. Protocol; 4.1. Preparation; 4.2. Duration; 4.3. Tip; 5. Step 1: Cloning the Target Gene into pNZ8048 and Transformation into L. lactis; 5.1. Overview; Amplification of the Target Gene by PCR; Restriction digestion; Preparation of pNZ vector; Restriction digestion; Ligation and pellet paint coprecipitation; Preparation of electrocompetent L. lactis; Transformation into electrocompetent L. lactis cells
  • 5.2. Tips
Dimensions
unknown
Edition
First edition.
Extent
1 online resource (693 p.)
Form of item
online
Isbn
9780128015216
Media category
computer
Media type code
c
Specific material designation
remote
System control number
  • (CKB)3710000000385534
  • (EBL)2009776
  • (SSID)ssj0001534340
  • (PQKBManifestationID)12551762
  • (PQKBTitleCode)TC0001534340
  • (PQKBWorkID)11494957
  • (PQKB)11088913
  • (MiAaPQ)EBC2009776
  • (EXLCZ)993710000000385534

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