Refactoring the nitrogen fixation gene cluster from Klebsiella oxytoca.

ABSTRACT: Bacterial genes associated with a single trait are often grouped in a contiguous unit of the genome known as a gene cluster. It is difficult to genetically manipulate many gene clusters because of complex, redundant, and integrated host regulation. We have developed a systematic approach to completely specify the genetics of a gene cluster by rebuilding it from the bottom up using only synthetic, well-characterized parts. This process removes all native regulation, including that which is undiscovered. First, all noncoding DNA, regulatory proteins, and nonessential genes are removed. The codons of essential genes are changed to create a DNA sequence as divergent as possible from the wild-type (WT) gene. Recoded genes are computationally scanned to eliminate internal regulation. They are or

SUBMITTER: Temme K 

PROVIDER: S-EPMC3345007 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature