If the donor molecule carries edits flanked by sequences homologous to the targeted locus ("homology arms"), the edits will be integrated as part of the repair process.
The pBALU-ext vector contains a cassette that confers kanamycin (Kan) resistance flanked by sequences homologous to the fosmid backbone pCC1FOS.
Firmicutes, which include many gram positive, spore forming species, and insect gut symbionts were represented in Atriplex by sequences homologous to Geobacillus (a genus associated with thermophiles), Clostridium and Sporobacter (two genera known for sulfur reducing activity) and Staphylococcus, which was particularly abundant in A. canescens.
The knockout cassette was made of the phleomycin resistance marker flanked by sequences homologous to regions within eqxS and eqxD.
Mutation cassettes contain a selection marker preceded by two transcriptional terminators and flanked by sequences homologous to the genomic target (see Figure 1).
As a result, the integrative DNA fragment containing P lac UV5 - lacI element linked to the sub-optimal SD sequence (GGGGGG), marked by cat gene and flanked by sequences homologous to the target sites of RSF1010 has been generated.
The stx locus in S. dysenteriae 1 is surrounded by DNA sequence homologous to lambdoid bacteriophage sequence; however, the toxin genes are not associated with a complete prophage genome (4, 5 ).
The PCR-amplified cassette is comprised of the aac(3 IV gene, conferring apramycin resistance, and an RK2 origin of transfer (oriT), and is flanked by DNA sequence homologous to regions immediately upstream and downstream of the gene to be deleted.
Exogenously provided DNA can be used as a repair template (by, e.g., possessing sequence homologous to the sequence neighboring the cut), promoting the integration of exogenous sequences at the target site via homology-directed repair (for review, see Gaj et al. 2013, 2014).
