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Genomic DNA of blood stages of WT or mutant parasites was isolated as described[36] and analyzed by LR-PCR using primer pair (p1, p2) 3258 (5'-TAAACCTATTTGAAGCTTTATAC) and 3259 (5'-CTTGTGGGAAATTACAATGAC) for correct integration of construct p5236FRT in the pf52/36 locus by double cross over integration.
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The system is an international lighthouse project showing the potential for aesthetical integration of constructed wetland systems in the built environment.
Clones that expressed the reporter shortly after integration were excluded from the analysis, because in these clones the expression could have resulted from rearrangement of the construct during integration or from integration of the construct in tandem arrays.
The integration of constructs post-implantation relies on the rapid formation of functional vasculature.
A major challenge for cartilage tissue engineering remains the proper integration of constructs with surrounding tissues in the joint.
Integration of constructs conveys carboxin resistance.
We show that the sdi1 locus is a useful site for targeted integration of constructs.
We establish the sdi1 of Z. tritici locus for targeted integration of constructs as single copies.
We also demonstrate that integration of constructs into the sdi1 locus by using carboxin has no detectable effect on virulence of Z. tritici on wheat.
Correct integration of constructs P1 and P2 were confirmed by Southern analyses of separated chromosomes and diagnostic PCR analyses (Fig. 3B).
Targeted insertions and cassette exchanges make possible the repeated integration of constructs into an identical DNA environment, thereby eliminating variations caused by position effects.
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