Transcription of the inserted sequences was driven by the immediate-early promoter of human cytomegalovirus in these vectors.
The ORF of ThbZIP1 was inserted into pROKII driven by CaMV35S promoter, designated as pROKII-ThbZIP1 (see Additional file 8: Table S5 for primers used).
Amplified products were digested and cloned into the yeast expression vector pFL61, in which the constitutive expression of the inserted gene was driven by the phosphoglycinecerate kinase (PGK) gene promoter.
Transcription of each insert was driven by the CMV promoter.
The inserted AtPAD4 gene was driven by the figwort mosaic virus subgenomic transcript (FMV-sgt) promoter [ 51], which exhibits strong, constitutive root expression.
Promoter regions of mtr-miR171h and MtNSP2, 900 bp as well as 1248 bp upstream of the start codon, respectively, were amplified from wild type (Medicago truncatula cv. Jemalong) genomic DNA and cloned into the binary vector pKGWFS7 containing GFP GUS fusion protein driven by the inserted vector.
For inducible expression, the G-Luc coding region was excised by digestion with HincII and BamHI and inserted into an inducible expression cassette driven by the Hsp70A promoter (Shao et al. 2007; Fig. 1b).
The ΔU3-HIV[MLV] vector carried a −418 to −40 U3 deletion and was obtained by inserting an internal ΔLNGFR expression cassette driven by the full MLV LTR into the pRRLsin-40.GFP vector [51].
To distinguish the homozygous noca-1∆ embryos from its wild-type and heterozygous siblings, we used MosSCI (Frøkjær-Jensen et al., 2008) to insert a reporter transgene (cytoplasmic mCherry driven by an epidermal promoter P lbp-1) at a site only 0.06 cM from the noca-1 locus, so that the wild-type noca-1+ is tightly linked to the mCherry reporter.
Nasal catheter inserted into the patient's nostrils, coolant driven by oxygen evaporates within the nasopharynx.
These changes are coherent with reduced loadings during the occlusion of the chewing molars, a movement driven by the masseter muscle inserted on the angular process.
