Gene Inactivation in the Cyanobacterium Synechococcus sp. PCC 7002 and the Green Sulfur Bacterium Chlorobium tepidum Using In Vitro-Made DNA Constructs and Natural Transformation

Niels-Ulrik Frigaard; Yumiko Sakuragi; Donald A Bryant

doi:10.1385/1-59259-799-8:325

Gene Inactivation in the Cyanobacterium Synechococcus sp. PCC 7002 and the Green Sulfur Bacterium Chlorobium tepidum Using In Vitro-Made DNA Constructs and Natural Transformation

Niels-Ulrik Frigaard, Yumiko Sakuragi, Donald A Bryant

Computational and RNA Biology

78 Citations (Scopus)

Abstract

Inactivation of a chromosomal gene is a useful approach to study the function of the gene in question and can be used to produce a desired phenotype in the organism. This chapter describes how to generate such mutants of the cyanobacterium Synechococcus sp. PCC 7002 and the green sulfur bacterium Chlorobium tepidum by natural transformation with synthetic DNA constructs. Two alternative methods to generate the DNA constructs, both performed entirely in vitro and based on the polymerase chain reaction (PCR), are also presented. These methods are ligation of DNA fragments with T4 DNA ligase, and megaprimer PCR.

Original language	English
Journal	Methods in Molecular Biology
Volume	274
Pages (from-to)	325-340
Number of pages	15
ISSN	1064-3745
DOIs	https://doi.org/10.1385/1-59259-799-8:325
Publication status	Published - 2004

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Gene Inactivation in the Cyanobacterium Synechococcus sp. PCC 7002 and the Green Sulfur Bacterium Chlorobium tepidum Using In Vitro-Made DNA Constructs and Natural Transformation. / Frigaard, Niels-Ulrik; Sakuragi, Yumiko; Bryant, Donald A.
In: Methods in Molecular Biology, Vol. 274, 2004, p. 325-340.

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title = "Gene Inactivation in the Cyanobacterium Synechococcus sp. PCC 7002 and the Green Sulfur Bacterium Chlorobium tepidum Using In Vitro-Made DNA Constructs and Natural Transformation",

abstract = "Inactivation of a chromosomal gene is a useful approach to study the function of the gene in question and can be used to produce a desired phenotype in the organism. This chapter describes how to generate such mutants of the cyanobacterium Synechococcus sp. PCC 7002 and the green sulfur bacterium Chlorobium tepidum by natural transformation with synthetic DNA constructs. Two alternative methods to generate the DNA constructs, both performed entirely in vitro and based on the polymerase chain reaction (PCR), are also presented. These methods are ligation of DNA fragments with T4 DNA ligase, and megaprimer PCR.",

author = "Niels-Ulrik Frigaard and Yumiko Sakuragi and Bryant, {Donald A}",

note = "Keywords: Chlorobium; Cyanobacteria; DNA, Bacterial; Gene Silencing; Mutagenesis; Transformation, Bacterial",

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doi = "10.1385/1-59259-799-8:325",

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AU - Frigaard, Niels-Ulrik

AU - Sakuragi, Yumiko

AU - Bryant, Donald A

N1 - Keywords: Chlorobium; Cyanobacteria; DNA, Bacterial; Gene Silencing; Mutagenesis; Transformation, Bacterial

PY - 2004

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N2 - Inactivation of a chromosomal gene is a useful approach to study the function of the gene in question and can be used to produce a desired phenotype in the organism. This chapter describes how to generate such mutants of the cyanobacterium Synechococcus sp. PCC 7002 and the green sulfur bacterium Chlorobium tepidum by natural transformation with synthetic DNA constructs. Two alternative methods to generate the DNA constructs, both performed entirely in vitro and based on the polymerase chain reaction (PCR), are also presented. These methods are ligation of DNA fragments with T4 DNA ligase, and megaprimer PCR.

AB - Inactivation of a chromosomal gene is a useful approach to study the function of the gene in question and can be used to produce a desired phenotype in the organism. This chapter describes how to generate such mutants of the cyanobacterium Synechococcus sp. PCC 7002 and the green sulfur bacterium Chlorobium tepidum by natural transformation with synthetic DNA constructs. Two alternative methods to generate the DNA constructs, both performed entirely in vitro and based on the polymerase chain reaction (PCR), are also presented. These methods are ligation of DNA fragments with T4 DNA ligase, and megaprimer PCR.

U2 - 10.1385/1-59259-799-8:325

DO - 10.1385/1-59259-799-8:325

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JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

Gene Inactivation in the Cyanobacterium Synechococcus sp. PCC 7002 and the Green Sulfur Bacterium Chlorobium tepidum Using In Vitro-Made DNA Constructs and Natural Transformation

Abstract

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