a new system is presented for the generation ofrecombinant bacillus subtilis strains without antibioticmarkers. this system is based on two plasmids constructed in escherichia coli. the first plasmid phm30contains an incomplete hisi gene, the last gene in thehistidine biosynthesis operon of b. subtilis and part ofthe genes yvca and yvcb of unkown function flankinghisi at the 3´-end. the spectin...

Recombinant adeno-associated virus (rAAV) vector holds promise for gene therapy. Despite a low frequency of chromosomal integration of vector genomes, recent studies have raised concerns about the risk of rAAV integration because integration occurs preferentially in genes and accompanies chromosomal deletions, which may lead to loss-of-function insertional mutagenesis. Here, by analyzing 347 rA...

Replication of retroviruses and retrotransposons depends on selecting a favorable chromosomal site for integration of their genomic DNA. Different retroelements meet this challenge by targeting distinctive chromosomal regions. Despite these differences, recent data hints at a common targeting mechanism-tethering of integration complexes to proteins bound at favorable sites.

Escherichia coli is used as a chassis for a number of Synthetic Biology applications. The lack of suitable chromosomal integration and expression loci is among the main hurdles of the E. coli engineering efforts. We identified and validated chromosomal integration and expression target sites within E. coli K12 MG1655 flagellar region 1. We analyzed five open reading frames of the flagellar regi...