A 44-residue hybrid peptide (CB (1-24)-Arg-Ser-Tyr-Tan (4-21)) incorporating 1-24 residues of cecropin B (CB) and 4-21 residues of thanatin (Tan) was designed and constructed. The CB-Tan gene was cloned into expressio...A 44-residue hybrid peptide (CB (1-24)-Arg-Ser-Tyr-Tan (4-21)) incorporating 1-24 residues of cecropin B (CB) and 4-21 residues of thanatin (Tan) was designed and constructed. The CB-Tan gene was cloned into expression plasmid pGEX-3X and expressed in E.coli BL21. The fusion protein was purified by affinity chromatography. After digested with enterokinase the gene product released with antibacterial activity and gave one band in Tricine-SDS-PAGE.展开更多
The aim of the present in vitro study was to assess the tumoricidal potential of the following natural peptides belonging to the Cecropin family, namely Cecropin A and B, on a series of tumour cell lines: MDA-MB-231 (...The aim of the present in vitro study was to assess the tumoricidal potential of the following natural peptides belonging to the Cecropin family, namely Cecropin A and B, on a series of tumour cell lines: MDA-MB-231 (breast adenocarcinoma) and M14K (human mesothelioma). The experimental results reveal that the cytotoxic effects of the two peptides depend on their concentration. Their efficiency is significant at 120 μM concentrations and it persists even at 60 μM concentrations. The effects were insignificant at 30 μM concentrations. On the other hand, the cytotoxic potential was not significantly dependant on the type of peptide but more on the type of tumour cell line used. The MDA MB 231 line cells were much more sensitive to the action of Cecropins A and B than the M14K line cells. The prospects brought about by this experimental research consist of the collection of in vitro experimental data on the tumoricidal potential of these natural cytotoxic peptides on tumour cells. This will enable specialists to develop future in vivo experimental models in order to test the antitumor effect of these cytotoxic peptides. The ultimate goal would be the discovery of agents with efficient antitumor properties, i.e. with maximum tumoricidal effects and minimum toxic side effects.展开更多
[ Objective ] This study aimed to investigate the antimicrobial activity of hybrid antimicrobial peptide buforin II-cecropin B. [ Method ] Gene fragment BC encoding the hybrid antimicrobial peptide Buforin II-Cecropin...[ Objective ] This study aimed to investigate the antimicrobial activity of hybrid antimicrobial peptide buforin II-cecropin B. [ Method ] Gene fragment BC encoding the hybrid antimicrobial peptide Buforin II-Cecropin B was synthesized by SOE-PCR with six primers designed according to the published amino acid sequences. Then, the BC gene fragment was ligated into pET32a vector and expressed in BI21. Antimierobial ability of the crude hybrid antimicrobial peptide was detected with the Oxford cup method. [ Result] The BC gene fragment was correctly amplified by SOE-PCR and ligated into pET32a vector, to construct the expression vector pET32-BC. The hybrid antimicrobial peptide buforin II-cecropin B was expressed in BI21 after induction by IPTG. The optimal induction time was 2 h. Antimicrobial activity assay suggested that the hybrid antimicrobial peptide possessed antimicrobial activity to Escherichia coli and Bacillus subtilis. [ Conclusion] The hybrid antimicrobial peptide buforin II-cecropin B exhibited antimicrobial activity to Gram-negative E. coli and Gram-positive B. subtilis strains.展开更多
To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the reco...To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.展开更多
文摘A 44-residue hybrid peptide (CB (1-24)-Arg-Ser-Tyr-Tan (4-21)) incorporating 1-24 residues of cecropin B (CB) and 4-21 residues of thanatin (Tan) was designed and constructed. The CB-Tan gene was cloned into expression plasmid pGEX-3X and expressed in E.coli BL21. The fusion protein was purified by affinity chromatography. After digested with enterokinase the gene product released with antibacterial activity and gave one band in Tricine-SDS-PAGE.
文摘The aim of the present in vitro study was to assess the tumoricidal potential of the following natural peptides belonging to the Cecropin family, namely Cecropin A and B, on a series of tumour cell lines: MDA-MB-231 (breast adenocarcinoma) and M14K (human mesothelioma). The experimental results reveal that the cytotoxic effects of the two peptides depend on their concentration. Their efficiency is significant at 120 μM concentrations and it persists even at 60 μM concentrations. The effects were insignificant at 30 μM concentrations. On the other hand, the cytotoxic potential was not significantly dependant on the type of peptide but more on the type of tumour cell line used. The MDA MB 231 line cells were much more sensitive to the action of Cecropins A and B than the M14K line cells. The prospects brought about by this experimental research consist of the collection of in vitro experimental data on the tumoricidal potential of these natural cytotoxic peptides on tumour cells. This will enable specialists to develop future in vivo experimental models in order to test the antitumor effect of these cytotoxic peptides. The ultimate goal would be the discovery of agents with efficient antitumor properties, i.e. with maximum tumoricidal effects and minimum toxic side effects.
基金Supported by the Research Project of Sichuan University of Science&Engineering(No.2011RC12,2014KY02)Scientific Research Foundation of the Education Department of Sichuan Province(NO.15ZA0222)
文摘[ Objective ] This study aimed to investigate the antimicrobial activity of hybrid antimicrobial peptide buforin II-cecropin B. [ Method ] Gene fragment BC encoding the hybrid antimicrobial peptide Buforin II-Cecropin B was synthesized by SOE-PCR with six primers designed according to the published amino acid sequences. Then, the BC gene fragment was ligated into pET32a vector and expressed in BI21. Antimierobial ability of the crude hybrid antimicrobial peptide was detected with the Oxford cup method. [ Result] The BC gene fragment was correctly amplified by SOE-PCR and ligated into pET32a vector, to construct the expression vector pET32-BC. The hybrid antimicrobial peptide buforin II-cecropin B was expressed in BI21 after induction by IPTG. The optimal induction time was 2 h. Antimicrobial activity assay suggested that the hybrid antimicrobial peptide possessed antimicrobial activity to Escherichia coli and Bacillus subtilis. [ Conclusion] The hybrid antimicrobial peptide buforin II-cecropin B exhibited antimicrobial activity to Gram-negative E. coli and Gram-positive B. subtilis strains.
基金Supported by Key Teachers Foundation of Education Office of Heilongjiang Province,2005(1055G005)
文摘To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.
