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It is of note that the three variants (104, 106, and 108), isolated by screening the libraries constructed by targeting the 3-HPA-binding site, have mutations in residues 442, 443, or 444 (Table 1).
Hairpin loop constructs that produce specific small-interfering RNAs (siRNAs; siNM23-H1 in text and figures) were designed and constructed by targeting GGAGAUCGGCUUGUGGUUU and GCUUCCGAAGAUCUUCUCA, two corresponding coding regions of human NM23-H1 anonspecificcontrolontrol siRNA GCGCGGGGCACGUUGGUGU (Yao et al, 2005).
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It can be regarded as the intersection of GRNs constructed by target predictions and GRNs constructed by co-expression models.
Therefore, the identification of the hub in the molecular network constructed by target genes of a particular miRNA helps us to understand biological and pathological roles of individual miRNAs.
We describe here two new mouse lines, constructed by gene targeting to the Rosa26 locus to facilitate Cre-mediated cell modification.
CIB1-deficient DT40 was constructed by serial gene targeting [26], [35].
A pSUPER-siAtg5 plasmid was constructed by inserting the targeting sequences 5′-GCAACTCTGGATGGGATTG-3′ of Atg5 into the pSUPER-basic vector by BglII and HindIII restrictions.
A bldD null mutant (SV77) was constructed by Redirect PCR targeting, and the bldD::apr mutant allele was moved into a new WT background by generalized transduction using the S. venezuelae-specific phage SV1.
A dual luciferase reporter plasmid analogous to the psiCH2-16T reporter used in Figure 2 was constructed by inserting target sequence derived from the cyclinE1 mRNA 3'UTR.
The cytokine layer was constructed by connecting target genes to corresponding measured cytokines.
As shown in Figure 2, a miRNA target protein network is constructed by miRNA-target interactions and PPI network data.
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