AIM: To construct @entity1 eosinophil-derived neurotoxin(hEDN) and @entity97 core protein (HBVc) eukaryotic fusion expression vector with a linker ( @entity69206 ) (3) between them to optimize the molecule folding, which will be used to inhibit @entity97 replication in vitro. METHODS: Previously constructed pcDNA3.1(-)/TR was used as a template. Linker sequence was synthesized and annealed to form dslinker, and cloned into pcDNA3.1(-)/TR to produce plasmid pcDNA3.1(-)/ @entity13210 -linker. Then the hEDN fragment was PCR amplified and inserted into pcDNA3.1(-)/ @entity13210 -linker to form pcDNA3.1(-)/ @entity60317 in which the effector molecule and the target molecule were separated by a linker sequence. pcDNA3.1(-)/ @entity60317 expression was identified by indirect immunofluorescence staining. Radioimmunoassay was used to analyse anti-HBV activity of pcDNA3.1(-)/ @entity60317 . Meanwhile, metabolism of cells was evaluated by @entity98 colorimetry. RESULTS: hEDN and HBVc eukaryotic fusion expression vector with a linker ( @entity69206 )(3) between them was successfully constructed. pcDNA3.1(-)/ @entity60317 was expressed in HepG @entity1 cells efficiently. A significant decrease of HBsAg concentration from pcDNA3.1(-)/ @entity60317 transfectant was observed compared to pcDNA3.1(-)/TR (P=0.036, P<0.05). @entity98 assay suggested that there were no significant differences between groups (P=0.08, P>0.05). CONCLUSION: Linker introduction enhances the inhibitory effect of @entity97 targeted ribonuclease significantly.

Inhibition of XXXX targeted ribonuclease enhanced by introduction of linker.

@entity1 :: ('9606', 'Species') :: ['human', '2.2.15'] @entity97 :: ('10407', 'Species') :: ['HBV'] @entity69206 :: ('p|SUB|G|(4|S', 'ProteinMutation') :: ['Gly(4)Ser'] @entity98 :: ('CHEBI:53233', 'Chemical') :: ['MTT'] @entity13210 :: ('85348', 'Gene') :: ['HBc'] @entity60317 :: ('440730', 'Gene') :: ['TNL']
@entity97 :: (10407,Species) :: ['HBV']
