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The ORF encoding the protease was amplified and cloned in pTZ57R/T vector and the resultant construct was designated as pTSP1.
The pETfliC plasmid construct was designated pGRP-1 (Figure 1A). Figure 1 Schematic presentation of the pET28c+ vector construct (6887 bp) containing the fli C gene responsible for the expression of rFliC designated as pGRP-1.
The Neospora caninum antigen (NcSRS2) expression cassette, consisting of the cytomegalovirus immediate-early promoter and NcSRS2 from N. caninum, was inserted into BmNPVΔbgp/AcGP64; this construct was designated BmNPVΔbgp/AcGP64/SRS2.
The construct was designated as pTIID.
The construct was designated fluorescent-conjoined sGC.
This construct was designated as pTIIDT.
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Final construct were designated as pWPTS-FL Ncl) and pWPTS-FL Nclcl) andordingly.
Cells incorporating the RNAi construct were designated K5KOAs.40 (KO cells); cells incorporating the inverse construct were Inv2 cells.
This expression constructed was designated as Luc-HNF4G.
The final constructs were designated pBD-TuMV-YC5 and pBD-TuMV-GFP, contained a complete cDNA copy of wild type TuMV and TuMV carrying GFP, respectively.
Sequence analysis of the transformants from the gene-replacement experiment confirmed that the INU1 ORF had been replaced by sequences encoding scFv; two of the resulting constructs were designated strains No. 6 (Km02-064) and No.7 (Km02-065) (Figs. 1b and 3b).
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