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Though numerous strategy options are available for achieving high expression levels of genes in Escherichia coli, not every gene can be efficiently expressed in this organism.
SDS-PAGE results showed that the α-subunit of NHBA and NHBAX could not be efficiently expressed in both recombinant E. coli strains.
We demonstrated this strategy on a promising antiviral candidate, Cyanovirin-N (CVN), which could not be efficiently expressed in any previously reported system.
These results showed that the ΔCP, a recombinant PCV2b capsid protein without nuclear localization signal sequence, could be efficiently expressed in Hansenula polymorpha, and used as a candidate antigen for the development of PCV2b vaccines.
Two recombinant antigens, fox zona pellucida protein subunit 3 (fZPC) and enhanced green fluorescent protein (EGFP) as control antigen, were inserted into thymidine kinase (TK) locus of the CHV genome and shown to be efficiently expressed in vitro.
A highly potent example is the thermophilic β-mannanase from Aspergillus niger BK01 (ManBK), which can be efficiently expressed in industrial yeast strains and is thus an attractive candidate for commercial utilizations.
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In these mice, N-cad was efficiently expressed in the E-cad expression domains.
SDS-PAGE analysis confirmed that the fusion protein HA1-2-fliC was efficiently expressed in an E. coli expression system (Fig. 3a and b).
The resulting fusion protein, HA1-2-fliC, was efficiently expressed in an Escherichia coli prokaryotic expression system, and Western blotting and TLR5-stimulating activity analysis confirmed that the HA1-2-fliC moiety could be faithfully refolded to take on the native HA and fliC conformations.
In the absence of IPTG, the D2-GFP fusion protein was efficiently expressed in E. coli transformed with pD2-GFP, while no GFP expression was detected in cells transformed with either pDSW207 or pDSW208 (Figure 4).
Although all three TbSTT3 proteins were efficiently expressed in wild-type yeast cells at similar levels (Supplementary Figure S1A), only expression of TbSTT3B and TbSTT3C, but not of TbSTT3A, could support yeast growth in the absence of endogenous STT3.
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