Abstract
In the post genome sequence era, metabolic engineering conducted by a number of genes is attracting great attention for not only basic research but also for application in production of beneficial materials for our lives. To carry this out, methods to handle a number of genes at a time are a prerequisite. However, the poor ability of Escherichia coli-based genetic methods to manipulate the reconstruction makes the steps timeconsuming and laborious. Bacillus subtilis, a gram-positive soil bacterium, develops natural competence in certain physiological conditions. In the competence, this bacterium can take up exogenous DNAs, and can recombine these DNAs with the genome if there is a homologous sequence. By using this trait, a novel method to carry out gene assembly quickly and with greatest fidelity using B. subtilis was developed. The method, named Ordered Gene Assembly in B. subtilis (OGAB), facilitates the connection of more than 10 DNA fragments in plasmid in one-step using an intrinsic B. subtilis plasmid transformation system. In this chapter, the principle of and actual protocol for the OGAB method are written. Moreover, two types of gene assembly in terms of metabolic engineering are also described.
| Original language | English |
|---|---|
| Title of host publication | Bioengineering: Principles, Methodologies and Applications |
| Publisher | Nova Science Publishers, Inc. |
| Pages | 153-168 |
| Number of pages | 16 |
| ISBN (Print) | 9781607417620 |
| Publication status | Published - 2010 |
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ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
Gene cluster or operon design by ordered gene assembly in Bacillus Subtilis (OGAB) method. / Tsuge, Kenji; Itaya, Mitsuhiro.
Bioengineering: Principles, Methodologies and Applications. Nova Science Publishers, Inc., 2010. p. 153-168.Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Gene cluster or operon design by ordered gene assembly in Bacillus Subtilis (OGAB) method
AU - Tsuge, Kenji
AU - Itaya, Mitsuhiro
PY - 2010
Y1 - 2010
N2 - In the post genome sequence era, metabolic engineering conducted by a number of genes is attracting great attention for not only basic research but also for application in production of beneficial materials for our lives. To carry this out, methods to handle a number of genes at a time are a prerequisite. However, the poor ability of Escherichia coli-based genetic methods to manipulate the reconstruction makes the steps timeconsuming and laborious. Bacillus subtilis, a gram-positive soil bacterium, develops natural competence in certain physiological conditions. In the competence, this bacterium can take up exogenous DNAs, and can recombine these DNAs with the genome if there is a homologous sequence. By using this trait, a novel method to carry out gene assembly quickly and with greatest fidelity using B. subtilis was developed. The method, named Ordered Gene Assembly in B. subtilis (OGAB), facilitates the connection of more than 10 DNA fragments in plasmid in one-step using an intrinsic B. subtilis plasmid transformation system. In this chapter, the principle of and actual protocol for the OGAB method are written. Moreover, two types of gene assembly in terms of metabolic engineering are also described.
AB - In the post genome sequence era, metabolic engineering conducted by a number of genes is attracting great attention for not only basic research but also for application in production of beneficial materials for our lives. To carry this out, methods to handle a number of genes at a time are a prerequisite. However, the poor ability of Escherichia coli-based genetic methods to manipulate the reconstruction makes the steps timeconsuming and laborious. Bacillus subtilis, a gram-positive soil bacterium, develops natural competence in certain physiological conditions. In the competence, this bacterium can take up exogenous DNAs, and can recombine these DNAs with the genome if there is a homologous sequence. By using this trait, a novel method to carry out gene assembly quickly and with greatest fidelity using B. subtilis was developed. The method, named Ordered Gene Assembly in B. subtilis (OGAB), facilitates the connection of more than 10 DNA fragments in plasmid in one-step using an intrinsic B. subtilis plasmid transformation system. In this chapter, the principle of and actual protocol for the OGAB method are written. Moreover, two types of gene assembly in terms of metabolic engineering are also described.
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UR - http://www.scopus.com/inward/citedby.url?scp=84896413456&partnerID=8YFLogxK
M3 - Chapter
AN - SCOPUS:84896413456
SN - 9781607417620
SP - 153
EP - 168
BT - Bioengineering: Principles, Methodologies and Applications
PB - Nova Science Publishers, Inc.
ER -