Transmembrane (TM) anchoring moieties were amplified by PCR, using the appropriate primers and templates (sequences available upon request) and were cloned, in-frame, as Xba-I/Pac-I digested PCR fragments, into the pCDNA 3.1-PS11-scFvFc expression vector.
The human Dysbindin-V5 (-V5 is tagged to C-terminal), Dysbindin-FLAG (-FLAG is tagged to N-terminal) and NF-YB moieties were amplified from a human brain cDNA library using PCR and subcloned into pcDNA3.1 expression vector (Invitrogen, Carlsbad, CA).
The Zera-KDEL moiety was amplified from pUC18Zera-ECFP [19] using two overlapping 5' primers designed to amplify the Zera sequence (lacking the signal peptide) preceded by an AscI restriction site, a sequence coding for Lumiotag (CCPGCC) and a sequence coding for five glycine residues (linker).
Bound virion (mRNA-peptide fusion) molecules were eluted and their RNA moiety was amplified and analyzed.
anisopliae was amplified.
mRNA (1.3 kb) was amplified by RT-PCR.
No gene was amplified in four isolates.
Then HBB site was amplified and sequenced.
Although ORFV DNA was amplified by real-time PCR, no other pathogen DNA tested was amplified.
But everything was amplified.
