Objective:To predict structure and function of translationally controlled tumor protein(TCTP)from Spirometraa mansoni by bioinformatics technology,and to provide a theoretical basis for further study.Methods:Open read...Objective:To predict structure and function of translationally controlled tumor protein(TCTP)from Spirometraa mansoni by bioinformatics technology,and to provide a theoretical basis for further study.Methods:Open reading frame(ORF)of EST sequence from Spirometra mansoni was obtained by ORE finder and was translated into amino acid residue by DNAclub.The structure domain was analyzed by Blast.By the method of online analysis tools:Protparam,InterProScan,protscale.SignalP-3.0,PSORTⅡ,BepiPred,TMHMM,VectorNT1 Suite 9 packages and Phyre2,the structure and function of the protein were predicted and analyzed.Results:The results showed that the EST sequence was Sm TCTP with 173 amino acid residues,theoretical molecular weight was 19 872.0 Da.The protein has the closest evolutionary status with Clonorchis sinensis.Schistosoma mansoni,and Schistosoma japonicum.Then it had no signal peptide site and transmembrane domain.Secondary structure of TCTP contained twoα-helices and eightβ-strands.Conclusions:Sm TCTP was a variety of biological functions of protein that may be used as a vaccine candidate molecule anti drug target.展开更多
Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.M...Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:Using the NCBI,EMBI,Expasy and other online sites, the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,transmembrane domain,epitope,topological structures of the protein sequences were predicted.And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.Results:’Hie target sequence was 1 132 hp length with a 681 hp biggest ORF encoding 226 amino acids protein with typical TSP conserved domain.It was confirmed as full length cDNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number:JF728872).The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da,respectively.Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni.it showed similarity of 59%and 59%, respectively.SeTSP16 contained four transmembrane domains(TM 1-4),intracellular N and C-termini,one short small extracellular loop and one large extracellular loop.Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.Conclusions:The full length cDNA sequences of SeTSP16 arc identified.It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.展开更多
Background The stage-specific expression of genes is o ne of the most characteristics of parasites. It has been found that a lot of gen es of Spriometra erinaceieuropaei are specifically expressed in pleroceroid in la...Background The stage-specific expression of genes is o ne of the most characteristics of parasites. It has been found that a lot of gen es of Spriometra erinaceieuropaei are specifically expressed in pleroceroid in large amount, but not expressed when the plerocercoid development into adult worm. The study is to screen other stage-specific ecpression genes of plerocerc oid of Spirtmetra erinceieuropaei.Methods RNA was separately extracted by acid guanidinium thiocyanate-phenol-chloroform from plerocercoids and adult worms of Spirometra erinaceieuropaei, DN A contaminated in the RNA was digested by RNase-free DNase. After the RNA was reverse transcripted to cDNA using T 12 MA, T 12 MC, T 12 MG and T 12 MT anchor-primers, PCR was done using the same T 12 MN and one random primer with α 35 S-dATP in the system. The PCR products were fractionated on an 8% denatured polyacrylamide gel. Differential bands of the plerocercoid found in the gel were cut out, amplified by PCR and sequenced. Northern hybridization was used to identify the stage-specific expression genes.Results Eleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization, after they were amplified by PCR. Fragments 1 (238 bp) and 2 (383 bp), were confirmed by Northern hybridization, as being expressed in the plerocercoid. However, fragment 3 (433 bp), was expressed in both the plerocercoid and the adult worm. Data from the 3 gene fragments underwent homological analysis in GenBank. The sequence which was homologous with fragments 1 and 2 was not found, but fragment 3 had high homology with many kinds of 28S rRNA. Conclusions The gene expressions of plerocercoids are different from adult worms because they live in different hosts. Two types of different gene fragments from the plerocercoid were found by mRNA differential display technique.展开更多
基金supported by Natural Science Foundation of China(Foundation No.81260254)
文摘Objective:To predict structure and function of translationally controlled tumor protein(TCTP)from Spirometraa mansoni by bioinformatics technology,and to provide a theoretical basis for further study.Methods:Open reading frame(ORF)of EST sequence from Spirometra mansoni was obtained by ORE finder and was translated into amino acid residue by DNAclub.The structure domain was analyzed by Blast.By the method of online analysis tools:Protparam,InterProScan,protscale.SignalP-3.0,PSORTⅡ,BepiPred,TMHMM,VectorNT1 Suite 9 packages and Phyre2,the structure and function of the protein were predicted and analyzed.Results:The results showed that the EST sequence was Sm TCTP with 173 amino acid residues,theoretical molecular weight was 19 872.0 Da.The protein has the closest evolutionary status with Clonorchis sinensis.Schistosoma mansoni,and Schistosoma japonicum.Then it had no signal peptide site and transmembrane domain.Secondary structure of TCTP contained twoα-helices and eightβ-strands.Conclusions:Sm TCTP was a variety of biological functions of protein that may be used as a vaccine candidate molecule anti drug target.
文摘Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:Using the NCBI,EMBI,Expasy and other online sites, the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,transmembrane domain,epitope,topological structures of the protein sequences were predicted.And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.Results:’Hie target sequence was 1 132 hp length with a 681 hp biggest ORF encoding 226 amino acids protein with typical TSP conserved domain.It was confirmed as full length cDNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number:JF728872).The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da,respectively.Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni.it showed similarity of 59%and 59%, respectively.SeTSP16 contained four transmembrane domains(TM 1-4),intracellular N and C-termini,one short small extracellular loop and one large extracellular loop.Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.Conclusions:The full length cDNA sequences of SeTSP16 arc identified.It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.
基金entialdisplaytechnThisworkwassupportedbytheScientificandTechnicalDepartmentofHebeiProvince (No 0 12 45512D)
文摘Background The stage-specific expression of genes is o ne of the most characteristics of parasites. It has been found that a lot of gen es of Spriometra erinaceieuropaei are specifically expressed in pleroceroid in large amount, but not expressed when the plerocercoid development into adult worm. The study is to screen other stage-specific ecpression genes of plerocerc oid of Spirtmetra erinceieuropaei.Methods RNA was separately extracted by acid guanidinium thiocyanate-phenol-chloroform from plerocercoids and adult worms of Spirometra erinaceieuropaei, DN A contaminated in the RNA was digested by RNase-free DNase. After the RNA was reverse transcripted to cDNA using T 12 MA, T 12 MC, T 12 MG and T 12 MT anchor-primers, PCR was done using the same T 12 MN and one random primer with α 35 S-dATP in the system. The PCR products were fractionated on an 8% denatured polyacrylamide gel. Differential bands of the plerocercoid found in the gel were cut out, amplified by PCR and sequenced. Northern hybridization was used to identify the stage-specific expression genes.Results Eleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization, after they were amplified by PCR. Fragments 1 (238 bp) and 2 (383 bp), were confirmed by Northern hybridization, as being expressed in the plerocercoid. However, fragment 3 (433 bp), was expressed in both the plerocercoid and the adult worm. Data from the 3 gene fragments underwent homological analysis in GenBank. The sequence which was homologous with fragments 1 and 2 was not found, but fragment 3 had high homology with many kinds of 28S rRNA. Conclusions The gene expressions of plerocercoids are different from adult worms because they live in different hosts. Two types of different gene fragments from the plerocercoid were found by mRNA differential display technique.
