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Both constructs were constructed in advance to reduce surgical procedure time and the frame diameters checked to avoid impingement of soft tissue.
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Briefly, targeting constructs were constructed using VelociGene technology as described previously (Valenzuela et al., 2003).
IRES-actinEGFP constructs were constructed by subcloning PICK1 cDNA, IRES cassette and actinEGFP cDNA into pcDNA3.1.
All expression constructs were constructed using Multisite Gateway recombination technology from Invitrogen (Carlsbad, CA).
The gfp and FLAG tagged constructs were constructed using Gibson Cloning on pCS2 vector backbones (Gibson et al., 2009).
The Dual promoter-GS-pOPINEE12G constructs were constructed by cloning the FcERα gene into the KpnI/PmeI sites of pOPINEE12G.
It was probably because all of the three shRNA constructs were constructed using the same expression vector, so their mixture was homogeneous, possibly suppressing CYP3A4 gene expression in an additive way.
Reporter constructs were constructed by insertion of two oligonucleotides: 5′-xxxTTCG-3′ and 5′-yyyTTGC-3′, where xxx and yyy represent the sense and antisense sequences of a selected 19-nt target respectively (Supplemental Figure 7D).
The respondents' views on these constructs are constructed by the subjective reality of their everyday life, which is influenced by the social realm that they are a part of.
The LRR1 overexpression construct was constructed by recombining the LRR1/pENTR/D-TOPO construct into the gateway-compatible Ubi-CAMBIA-1300 (Chern et al. 2005).
The pET22b-mMMP12-CPD pET22b-mMMP12-CPD pET22b-mMMP12-CPDplifying the construct domain of mouse MMP12 (amino acids 29–267) using pET41a-mMMP12 as a template using primers #15 and #16.
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