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Cloning, expression and purification of Schistosoma japonicum ribosomal protein S4 as well as the preliminary study of the diagnostic value of the recombinant protein / 中华预防医学杂志
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-291488
Responsible library: WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To express and purify Schistosoma japonicum ribosomal protein S4(SjRPS4) in Escherichia coli, and assess its value in immunodiagnosis of Schistosomiasis japonica.</p><p><b>METHODS</b>Gene fragment of SjRPS4 was amplified by screening the cercaria cDNA library of Schistosoma japonicum. The target gene was cloned into the expressive vector pQE30 and transformed into E. coli M15. The recombinant protein expression was induced by isopropylthio-β-D-galactoside (IPTG). This fusion protein was purified by Ni(2+)-NTA chromatography and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and ELISA.</p><p><b>RESULTS</b>The plasmid pQE30/SjRPS4 was constructed successfully and expressed a SjRPS4 fusion protein in E. coli as showing a single special band on SDS-PAGE gel at Mr 30 × 10(3) position. It reached a purity of above 90% after purification. The Western blot result confirmed that the recombinant protein could specifically react with the serum samples from patients of schistosomiasis. Detecting the serum of Schistosomiasis japonica patients by ELISA, the sensitivity and specificity of the ELISA method were 90.91% (70/77) and 92.59% (25/27), the positive rate of recombinant protein expression was 67.30% (70/104). There was no cross-reaction with paragonimiasis patients' serum.</p><p><b>CONCLUSION</b>Protein SjRPS4 was successfully cloned and expressed, and it was confirmed that SjRPS4 antibodies were valuable in the diagnosis of Schistosomiasis japonica.</p>
Subject(s)
Full text: Available Health context: Neglected Diseases Health problem: Helminthiasis / Neglected Diseases / Schistosomiasis / Zoonoses Database: WPRIM (Western Pacific) Main subject: Plasmids / Ribosomal Proteins / Schistosoma japonicum / Blood / Schistosomiasis japonica / Recombinant Proteins / Enzyme-Linked Immunosorbent Assay / Antibodies, Helminth / Molecular Sequence Data / Gene Library Type of study: Diagnostic study Limits: Animals / Humans Language: Chinese Journal: Chinese Journal of Preventive Medicine Year: 2010 Document type: Article
Full text: Available Health context: Neglected Diseases Health problem: Helminthiasis / Neglected Diseases / Schistosomiasis / Zoonoses Database: WPRIM (Western Pacific) Main subject: Plasmids / Ribosomal Proteins / Schistosoma japonicum / Blood / Schistosomiasis japonica / Recombinant Proteins / Enzyme-Linked Immunosorbent Assay / Antibodies, Helminth / Molecular Sequence Data / Gene Library Type of study: Diagnostic study Limits: Animals / Humans Language: Chinese Journal: Chinese Journal of Preventive Medicine Year: 2010 Document type: Article
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