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This specific integration resulted in complete abolishment of heparanase mRNA expression and enzymatic activity, as revealed by PCR analysis applying 3 different pairs of primers and lack of heparanase enzymatic activity in blood samples and tissue extracts of the Hpse-KO mice.
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We analysed this parasite population by Southern blot, using a probe diagnostic for the specific integration event.
The minimum time indicated was the specific integration time that fulfilled the defects visibilities compared with film.
Genomic PCR amplification was used to validate the specific integration vector pMoC-eGFP into the Mosdi1 locus of M. oryzae by a set of primers GL728/GL565.
Analyses of gene-edited FA-iPSCs confirmed the specific integration of FANCA in the AAVS1 locus in all tested clones.
All HEK293 G418-resistant derived clones (daughter clones) analysed showed the specific integration of the targeting construct and lack unspecific random integration.
These PCR assays target the specific integration of IS 711 elements within the genome of the respective Brucella species or biovar.
All of these effects can vary according to the specific integration event and would, therefore, be unique to each independent transgenic line.
This suggests that the strength/potential of a specific integration site is linked to a certain promoter - and is not necessarily supporting any integrated expression cassette.
The SXT element from Vibrio cholerae was also able to integrate in a tandem fashion in E. coli into its specific integration site [18].
Reference time steps were identified for specific integration scheme.
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