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While transient transfection of Babesia spp. has been reported previously, stable integration of exogenous genes in Babesia has proven difficult.
This shows that the developed system is very useful for the markerless integration of exogenous genes into the chromosome of gram-negative eubacteria.
In this study, we establish a novel strategy for locus-specific integration of exogenous DNA via carboxin-resistance reconstitution by a point-mutation (H245L) on succinate dehydrogenase subunit Mosdi1.
As sdi1 R locus has been recommended for locus-specific integration of exogenous DNA in study of protein localization and gene complementation in filamentous phytopathogens (Kilaru et al. 2015, 2009; Shima et al. 2009), thus, a locus specific integration vector pMoC-eGFP based on mutated Mosdi1 sequence was constructed.
Repair of DSBs by HR allows integration of exogenous sequences at target site.
Viral vectors have been effectively used to promote the integration of exogenous DNA into T cells.
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The reason for the non-linear dependence of expression on copy number is unclear but one possibility is that it involves gene silencing caused by integration sites of exogenous genes.
In spite of the genomic integration of an exogenous gene, BDNF MSCs did not present any structural aberration in the chromosomes.
These transgenic or viral based strategies, however, involve integration of the exogenous DNA into random genomic loci that can modulate the promoter activity, depending on the epigenetic state around the integration site.
For example, Balbontin have recently discovered that small RNAs could mediate prophage insertion and consequently integration of carried exogenous DNA [83].
To confirm the cellular integration of the exogenous gp91phox 221-C proteintonthethe plasma membrane a set of experiments has been performed using freshly produced gp91phox 221-C proteoliposomes.
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