To rapidly and economically obtain knowledge about adult stage Schistosoma japonicum (Chinese strain) expressed genes using expressed sequence tag (EST) Methods A directional cDNA library constructed from Schisto...To rapidly and economically obtain knowledge about adult stage Schistosoma japonicum (Chinese strain) expressed genes using expressed sequence tag (EST) Methods A directional cDNA library constructed from Schistosoma japonicum (Chinese strain ) adult stage RNA was used to generate expressed sequence tags(ESTs) These were compared against an EMBL parasites database and GENBANK database by BLASTn and BLASTx Results A total of 314 phage clones were randomly selected for generating expressed sequence tags(ESTs) From these clones, 132 EST quality sequence were obtained Among these EST quality sequences,113 ESTs were successfully submitted to the dbEST at GenBanK A total of 7 6% of these EST quality sequences were previously identified sequence of Schistosoma japonicum, while 4 5% were putatively identified sequences of Schistosoma japonicum A total of 23 5% of these EST quality sequences were putatively identified sequence of Schistosoma mansoni or other organisms 57 6% had no matches in the database and were classified as unknown sequences Most ESTs with the putative protein identified belonged to housekeeping proteins Information about several interesting genes was found Conclusion Partial cDNA sequencing to generate expressed sequence tags (ESTs) has the potential to rapidly and economically increase our knowledge about adult stage Schistosoma japonicum(Chinese strain) expressed genes展开更多
Abstract Objective To develop the vaccine of Chinese Schistosomiasis japonicum, we try to prepare the 23kDa membrane protein of Schistosoma japonicum Chinese strain with the gene cloning techniques. Methods A pair ...Abstract Objective To develop the vaccine of Chinese Schistosomiasis japonicum, we try to prepare the 23kDa membrane protein of Schistosoma japonicum Chinese strain with the gene cloning techniques. Methods A pair of primers P1 and P2 was systhesized according to the DNA sequence of the 23kDa membrane protein of Schistosoma mansoni, a BamH1 site was added at 5' end of the primer P1 and the Sall site was added at the 5' end of the primer P2. The gene DNA fragment of the 23kDa membrane protein (SjC23) of Schistosoma japonicum was amplified from the cDNA library of Schistosoma japonica by PCR, the purified target DNA fragment was inserted into the vector pUC18/19 to form the recombinant, and sequenced in Livopool University, UK and Fudan Universtiy, China respectively. The DNA sequence was analyzed with Dnasis software, and the amino sequence was deduced with the SWISS PORT software. Results The size of DNA of 23kDa membrane protein of Schistosoma japonica Chinese strain (SjC23) was 657bp, and it was the same size as that of Sm23 and Sj23 Philippine strain. The DNA sequence of Sj23 Chinese strain (SjC23) was 100% in homology with the SjC23 Philippine strain, and 79.5% in homology with Sm23. The deduced amino acid sequence of SjC23 was 84% in homology with the Sm23, and 100% in homology with Philippine strain. There were two hydrophilic domains in the SjC23, one was located at the N terminal (amino acid 36-56), and another was at the C terminal (amino acid 108-183). Conclusions The gene of the 23kDa membrane protein of Schistosoma japonica Chinese strain has been cloned, and this work has laid the foundation for the development of the vaccine of Schistosoma japonica Chinese strain.展开更多
基金ThisprojectwassupportedbyNationalNaturalScienceFoundationofChina (No 30 0 70 683 )
文摘To rapidly and economically obtain knowledge about adult stage Schistosoma japonicum (Chinese strain) expressed genes using expressed sequence tag (EST) Methods A directional cDNA library constructed from Schistosoma japonicum (Chinese strain ) adult stage RNA was used to generate expressed sequence tags(ESTs) These were compared against an EMBL parasites database and GENBANK database by BLASTn and BLASTx Results A total of 314 phage clones were randomly selected for generating expressed sequence tags(ESTs) From these clones, 132 EST quality sequence were obtained Among these EST quality sequences,113 ESTs were successfully submitted to the dbEST at GenBanK A total of 7 6% of these EST quality sequences were previously identified sequence of Schistosoma japonicum, while 4 5% were putatively identified sequences of Schistosoma japonicum A total of 23 5% of these EST quality sequences were putatively identified sequence of Schistosoma mansoni or other organisms 57 6% had no matches in the database and were classified as unknown sequences Most ESTs with the putative protein identified belonged to housekeeping proteins Information about several interesting genes was found Conclusion Partial cDNA sequencing to generate expressed sequence tags (ESTs) has the potential to rapidly and economically increase our knowledge about adult stage Schistosoma japonicum(Chinese strain) expressed genes
文摘Abstract Objective To develop the vaccine of Chinese Schistosomiasis japonicum, we try to prepare the 23kDa membrane protein of Schistosoma japonicum Chinese strain with the gene cloning techniques. Methods A pair of primers P1 and P2 was systhesized according to the DNA sequence of the 23kDa membrane protein of Schistosoma mansoni, a BamH1 site was added at 5' end of the primer P1 and the Sall site was added at the 5' end of the primer P2. The gene DNA fragment of the 23kDa membrane protein (SjC23) of Schistosoma japonicum was amplified from the cDNA library of Schistosoma japonica by PCR, the purified target DNA fragment was inserted into the vector pUC18/19 to form the recombinant, and sequenced in Livopool University, UK and Fudan Universtiy, China respectively. The DNA sequence was analyzed with Dnasis software, and the amino sequence was deduced with the SWISS PORT software. Results The size of DNA of 23kDa membrane protein of Schistosoma japonica Chinese strain (SjC23) was 657bp, and it was the same size as that of Sm23 and Sj23 Philippine strain. The DNA sequence of Sj23 Chinese strain (SjC23) was 100% in homology with the SjC23 Philippine strain, and 79.5% in homology with Sm23. The deduced amino acid sequence of SjC23 was 84% in homology with the Sm23, and 100% in homology with Philippine strain. There were two hydrophilic domains in the SjC23, one was located at the N terminal (amino acid 36-56), and another was at the C terminal (amino acid 108-183). Conclusions The gene of the 23kDa membrane protein of Schistosoma japonica Chinese strain has been cloned, and this work has laid the foundation for the development of the vaccine of Schistosoma japonica Chinese strain.
