Download BacLoxP Baculovirus Expression System

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TABLE OF CONTENTS
A. flEXi Coli kit contents
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Reagents to be supplied by user (see also section D. Protocols)
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B. Escherichia coli – a premier laboratory workhorse
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B.1. Introduction
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C. A Novel Approach to Multigene Applications
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C.1. The Toggle Concept – a Primer
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C.2. Transfer vectors: Acceptor-Donor recombineering system.
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C.3. Generating multi-gene expression cassettes
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C.3.1. Creating individual gene expression cassettes
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C3.1.1. Standard Approach
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C3.1.2 Inserts with Other Unique Restriction Sites
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C.3.2. Multi-gene construction via Toggle multiplication
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C.3.3. Multi-gene construction using Cre-Lox recombination
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C.4. Introducing additional control elements
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D. Protocols
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D.0 Introductory remarks
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D.1 Cloning into TogColi transfer vectors
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D.2 Conversion of donors into conditional donors
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D 2.1 Protocol 1. Conversion into conditional oris
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D.3 Concatenation of genes (gene cassettes) by Toggling
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D 3.1 Protocol 2. Adding gene cassettes through toggling
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D.4 Deconstruction of gene cassette assemblies
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D 4.1 Protocol 3. Disassembling multi-gene construct to regenerate individual expression boxes
D.5 Cre-LoxP reaction of Acceptors and Donors
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D 5.1. Protocol 4: Cre-LoxP fusion of Acceptors and Donors
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D 5.2. Protocol 5. Deconstruction of fusion vectors by Cre
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D.6. Expression of multi-gene constructs
E. Appendix
E.1. Preparing chemically competent cells
flEXiColi System
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ATG:biosynthetics GmbH