The fragile antibody leads to a great challenge as a scaffold to fabricate the luminescent metal nanoclusters using one-pot method.This study presents a stable single-chain anti-body(scFv57R-ATS)for the fabrication of...The fragile antibody leads to a great challenge as a scaffold to fabricate the luminescent metal nanoclusters using one-pot method.This study presents a stable single-chain anti-body(scFv57R-ATS)for the fabrication of luminescent gold nanoclusters(AuNCs@scFv57R-ATS)and a quick,sensitive rabies virus detection in living cells.In this paper,AuNCs@scFv57R-ATS was designed to specifically recognize antigen RV in modified HeLa cells,which promoted the demonstration of metal nanocluster fluorescent probes for antigen targeting and therapy.展开更多
Targeting the N gene of rabies virus (RV), four shRNA expression plasmids were designed and constructed based on the vector pRNATU6.3-Hygro that expresses fusion protein with GFP as a reporter gene. Four cell strains ...Targeting the N gene of rabies virus (RV), four shRNA expression plasmids were designed and constructed based on the vector pRNATU6.3-Hygro that expresses fusion protein with GFP as a reporter gene. Four cell strains (N1, N2, N3, N4) expressing the short hairpin RNAs (shRNA) were obtained after the plasmids were transfected into BHK-21 cells and screened under the pressure of Hygromycin B (300 μg/mL). These cell strains were infected with 100× the TCID 50 of rabies virus CVS-11 strain, and the viral replication was quantified at 24, 48, 72 and 96 hours by directed immunofluorescence assay (DFA), real-time PCR, and the 50% tissue culture infective dose (TCID 50 ). The results showed variable inhibition of viral replication, with BHK-N2 being the most effective strain (99% inhibition). There was close correspondence between results using the three methods of evaluation. The shRNA-mediated inhibition persisted to at least 96 hours after infection. Effective inhibition of replication of RV in BHK-21 cells was achieved by siRNA targeting the N gene, with N 2 , aimed at the region starting at position 701 of the gene, being the most potent.展开更多
文摘The fragile antibody leads to a great challenge as a scaffold to fabricate the luminescent metal nanoclusters using one-pot method.This study presents a stable single-chain anti-body(scFv57R-ATS)for the fabrication of luminescent gold nanoclusters(AuNCs@scFv57R-ATS)and a quick,sensitive rabies virus detection in living cells.In this paper,AuNCs@scFv57R-ATS was designed to specifically recognize antigen RV in modified HeLa cells,which promoted the demonstration of metal nanocluster fluorescent probes for antigen targeting and therapy.
文摘Targeting the N gene of rabies virus (RV), four shRNA expression plasmids were designed and constructed based on the vector pRNATU6.3-Hygro that expresses fusion protein with GFP as a reporter gene. Four cell strains (N1, N2, N3, N4) expressing the short hairpin RNAs (shRNA) were obtained after the plasmids were transfected into BHK-21 cells and screened under the pressure of Hygromycin B (300 μg/mL). These cell strains were infected with 100× the TCID 50 of rabies virus CVS-11 strain, and the viral replication was quantified at 24, 48, 72 and 96 hours by directed immunofluorescence assay (DFA), real-time PCR, and the 50% tissue culture infective dose (TCID 50 ). The results showed variable inhibition of viral replication, with BHK-N2 being the most effective strain (99% inhibition). There was close correspondence between results using the three methods of evaluation. The shRNA-mediated inhibition persisted to at least 96 hours after infection. Effective inhibition of replication of RV in BHK-21 cells was achieved by siRNA targeting the N gene, with N 2 , aimed at the region starting at position 701 of the gene, being the most potent.
