Abstract
Riemerella anatipestifer is a Gram-negative, pathogenic bacterium, which is harmful to poultry. However, the genomic islands (GIs) in R. anatipestifer are not well-studied. In this study, a 10K genomic island was predicted by the bioinformatics analysis of R. anatipestifer ATCC 11845, which is widely found in other R. anatipestifer genomes. We had first reported the genomic island integration and excision function in R. anatipestifer. We successfully constructed the integration plasmid by using the integrase and 53 bp attP elements. The 10K GI was found integrated at the 53 bp attB located in the Arg-tRNA of the R. anatipestifer RA-YM chromosome. We identified an integrase that helped in the precise integration and excision in R. anatipestifer and elucidated the molecular mechanism of the 10K genomic island integration and excision. Furthermore, we provided a new method for the gene expression and construction of complementary strain.
Highlights
Genomic islands (GIs) are the segments of genomic DNA, which were originally known as pathogenicity islands (PAIs) in the 1980s
Our results suggested that the 10K GI integrase has both integration and excision functions
The GIs share several features: (1) large DNA segments, usually between 10 and 200 kb (Song et al, 2012), (2) the GC content usually differ from the rest of the chromosome, FIGURE 3 | Integrase mediated integration and excision. (A) Primer designed for integrase cloned. (B) The vector construction and integration mechanism
Summary
Genomic islands (GIs) are the segments of genomic DNA, which were originally known as pathogenicity islands (PAIs) in the 1980s. The GIs were first reported by Hacker while studying the virulence mechanisms in genetics and evolution of Escherichia coli (Hacker et al, 1997). They play a key role in prokaryotic genome plasticity. GIs are integrated into chromosomal loci, such as transfer RNA genes and protein-coding genes while retaining various cargo genes that potentially bestow novel functions on the host organism. The GIs have integrase/recombinase genes that facilitate chromosome integration and excision (Hsiao et al, 2005). The excisionase will cut off the direct repeat sequence at both the ends of the GIs and form attB and attP (Williams, 2002)
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