| RRC ID |
34504
|
| 著者 |
Koma D, Yamanaka H, Moriyoshi K, Ohmoto T, Sakai K.
|
| タイトル |
A convenient method for multiple insertions of desired genes into target loci on the Escherichia coli chromosome.
|
| ジャーナル |
Appl Microbiol Biotechnol
|
| Abstract |
We developed a method to insert multiple desired genes into target loci on the Escherichia coli chromosome. The method was based on Red-mediated recombination, flippase and the flippase recognition target recombination, and P1 transduction. Using this method, six copies of the lacZ gene could be simultaneously inserted into different loci on the E. coli chromosome. The inserted lacZ genes were functionally expressed, and β-galactosidase activity increased in proportion to the number of inserted lacZ genes. This method was also used for metabolic engineering to generate overproducers of aromatic compounds. Important genes of the shikimate pathway (aroF (fbr) and tyrA (fbr) or aroF (fbr) and pheA (fbr)) were introduced into the chromosome to generate a tyrosine or a phenylalanine overproducer. Moreover, a heterologous decarboxylase gene was introduced into the chromosome of the tyrosine or phenylalanine overproducer to generate a tyramine or a phenethylamine overproducer, respectively. The resultant strains selectively overproduced the target aromatic compounds. Thus, the developed method is a convenient tool for the metabolic engineering of E. coli for the production of valuable compounds.
|
| 巻・号 |
93(2)
|
| ページ |
815-29
|
| 公開日 |
2012-1-1
|
| DOI |
10.1007/s00253-011-3735-z
|
| PMID |
22127754
|
| MeSH |
Bacteriophage P1 / genetics
Chromosomes, Bacterial*
Escherichia coli / genetics*
Genes, Reporter
Genetic Engineering / methods*
Metabolic Engineering / methods
Mutagenesis, Insertional / methods*
Recombination, Genetic
Transduction, Genetic
beta-Galactosidase / genetics
beta-Galactosidase / metabolism
|
| IF |
3.53
|
| 引用数 |
19
|
|
WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 1170 |
