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Open AccessJournal ArticleDOI

An improved Escherichia coli donor strain for diparental mating

Sabrina Thoma, +1 more
- 01 May 2009 - 
- Vol. 294, Iss: 2, pp 127-132
TLDR
This improved method uses a new donor strain, E. coli ST18, a hemA deletion mutant mutant defective in tetrapyrrole biosynthesis, which means that counterselection of the Escherichia coli donor strain is not required.
Abstract
We present a new method for diparental mating with the outstanding advantage that counterselection of the Escherichia coli donor strain is not required. This improved method uses a new donor strain, E. coli ST18, a hemA deletion mutant defective in tetrapyrrole biosynthesis. The hemA mutation can be complemented by addition of 5-aminolevulinic acid. Therefore, counterselection is carried out only using standard media and growth conditions optimal for the recipient strain. Consequently, recipient strains are isolated in a significantly shorter period.

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References
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Book

Molecular Cloning: A Laboratory Manual

TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
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One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products

TL;DR: A simple and highly efficient method to disrupt chromosomal genes in Escherichia coli in which PCR primers provide the homology to the targeted gene(s), which should be widely useful, especially in genome analysis of E. coli and other bacteria.
Journal ArticleDOI

A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria

TL;DR: In this paper, a new vector strategy for the insertion of foreign genes into the genomes of gram negative bacteria not closely related to Escherichia coli was developed, which can utilize any gram negative bacterium as a recipient for conjugative DNA transfer.
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TL;DR: The Enteric Bacterial Cell and the Age of Bacteria Variations on a Theme by Escherichia is described.
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A short course in bacterial genetics

TL;DR: This second edition of Molecular Cloning reflects more comprehensive coverage of topics previously included in the first edition as well as the addition of new chapters, such as those on oligonucleotide probes and mutagenesis, in vitro amplification by the polymerase chain reaction, expression of cloned genes in Escherichia coli and mammalian cells, and analysis of proteins ex­ pressed from cloning genes.
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