The current design of the expression vector
contributes 4.2 kDa at the amino terminus, which is common to all peptides, and 1.5 kDa at the carboxy terminus (for those sequences that do not terminate internally), which may aid in satisfying any postulated "minimum size" requirement for protein accumulation.
Then, it was sub-cloned using a ligation-restriction step into the prokaryotic expression vector
pET-26b+(5360 bps) by NcoI/HindIII double digestion (Figure 6D).
To apply the cumate-inducible transgene expression system, the MyoD gene, an important regulator of muscle differentiation, was inserted into the piggyBac cumate-inducible expression vector
, and stably transfected DF1 cells were established following transfection and puromycin selection.
In our previous experiments, the BnPIP-like gene fragment was fused into prokaryotic expression vector
pET30a and then induced into Escherichia coli BL21 (DE), in order to operate the prokaryotic expression (Xu et al., 2014).
Unlike our study, Peerayeh SN and colleagues (25) used from prokaryotic expression vector
pET28a and like present study, E.
The ORFs of the two chitinase genes, LbCHI31 and LbCHI32, were amplified using the corresponding primers and digested with double enzymes (Table 1), ligated into the expression vector
of pET52b, and transferred into E.
Expression plasmid pTRP-CK-M was constructed by insertion of the DNA fragment prepared by digesting the pBluescript 11 SK(-)-CK-M with EcoRI/ BamHI, which encodes the human CK M-type subunit, into expression vector
pTRP (3) that had been digested with EcoRl/BamH1.
The secretin domain of PilQ (that's mean Pil[Q.sub.380-706]) was chosen as a new antigen and designed into expression vector
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The expression vector
of recombinant SPLUNC1 protein was constructed by inserting full length SPLUNC1 open reading frame (ORF) into pPIC9K plasmid (Invitrogen, China).
pHT43 (catalog number PBS002) Bacillus expression vector
was purchased from MoBiTec GmbH Germany.
The restriction site for NdeI enzyme was added before the start codon of confirmed gene sequence and complete open reading frame was ligated into pET22b (+) expression vector
using NdeI and EcoRI restriction sites for both the gene and the vector.