The RNAi vector of OsDR10 was constructed by inserting a fragment of cDNA digested from cDNA clone BI71N2 using restriction enzymes KpnI and BamHI into the pDS1301 vector (Figure S2B) [37].
A luciferase reporter construct, psi-FGFR1, was constructed by inserting a fragment of FGFR1 3′-UTR that included miR-133a/b and miR-214 binding sites into the luciferase reporter vector psi-CHECK-2.
The strain MK-ER-Ndt80 (inducible Ndt80 by estradiol, see Carlile et al [ 29]), was made by inserting a fragment of the plasmid PKB80 into the ura3 locus (selecting for Ura+ transformants).
To investigate whether the contribution of miR-204 to the enhanced killing of GTL-16 upon treatment with 5-fluorouracil or oxaliplatin occurs, at least in part, through the targeting of Bcl-2 expression, we inserted a fragment of 541 base pair of the Bcl-2 3′-UTR conthreeng three miR-204 binding sequences downstream of the luciferase coding regions (Supplementary Figure 3e f).
To test in vitro whether the methylation of the P2 promoter affects IGF1 gene expression, we used a reporter gene construct made by inserting a fragment of IGF1 promotertextendinging from −1,014 to +64 bp of the TSS - thus containing 7 of the studied CGs - into a promoter-less CpG-free luciferase expression plasmid.
The full-length human RIP3 cDNA was cloned into pFastBac HT B plasmids (Invitrogen, Grand Island, NY, USA), which was modified by inserting a fragment of DNA sequence encoding GST into the upstream of the multiple cloning sites of the primary plasmid.
Although whole clones are employed in some hybridisation studies [ 3], usually only the insert or a fragment of it is used as a probe.
The reporter plasmid, a firefly luciferase reporter construct psiCHECK-2 (Promega, WI) inserted with a fragment of the 3′-UTR of rat Drd1 mRNA containing the putative miR-382 binding sequence.
These observations do not appear to be directly related to the IRES element itself, as deletion variants appear to arise at an equivalent rate following passage of virus with a similar total insert size, but containing a fragment of LUC in place of an RhPV sequence (data not shown).
The goals are to insert a DNA fragment of interest into a receiving vector plasmid, transform the plasmid into E. coli, recover the plasmid DNA, and check for correct insertion events.
And in "Alligator Boogaloo," a blues phrase or a fragment of one was inserted in an abstract improvisation like a streak of lush color in a dot-matrix design.
