Identifying the insertion event has previously met with varying degrees of success, from a cleanly disrupted gene with minimal deletion of nucleotides at the insertion point to multiple-copy insertion events and large deletions of chromosomal regions.
The resulting assemblies were used to confirm the orientation, bp deletion, and plasmid insertion copy number where applicable (not successful for multiple copy plasmid insertions) by alignment to the reference sequence Fusarium graminearum PH-1 genome and the plasmid sequence within Geneious version 7.1.6 created by Biomatters (available from http://www.geneious.com/) using LASTZ [ 39].
The MmmSC genome is larger than that of Mmc, mostly due to gene duplications, and the insertion of multiple copies of Insertion Sequences.
In fact, this phenomenon was demonstrated for the GR-541 plasmid that contains multiple copies of insertion sequences and other mobile genetic elements within its scaffold (14 ).
Compared to those of other strictly intracellular bacteria, the C. burnetii genome harbors multiple copies of insertion sequence (IS) elements that are probably involved in genomic plasticity [ 13], but it possesses fewer pseudogenes, suggesting a recent genome reduction event [ 10].
As is common with the mouse microinjection method, we observed a high germ-line transmission rate, multiple site insertion, rearrangement, and multiple DNA-copy insertion using LB-SMGT in pigs.
This stable KD approach was used because of the multiple copy of E6/E7 insertion in the genome of HeLa cells making traditional knock-out experiments non-viable.
In "stable" transfections, the transfected DNA typically inserts as an array of multiple copies; the insertions occur at undefined and irreproducible chromosomal sites, the copy number varies idiosyncratically, and the multiple copies are in both orientations.
This is in contrast with the traditional production of Tg mice which is based on random insertion of multiple copy numbers of transgenes [ 2].
Moreover, nosocomial S. epidermidis strains typically harbour multiple copies of the insertion sequence element IS256 in their genomes, which contribute to genetic adaptation during infection [ 6].
