Cloning and expression of the succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum Chinese strain in E. coli / 中南大学学报(医学版)
Journal of Central South University(Medical Sciences)
; (12): 458-463, 2006.
Article
en Zh
| WPRIM
| ID: wpr-813671
Biblioteca responsable:
WPRO
ABSTRACT
OBJECTIVE@#To clone the full-length gene encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) Chinese strain and express it in Escherichia coli.@*METHODS@#According to the published incomplete EST (BU804141) of SjSDISP and the sequence of multiclone sites of lambda gt11 vector, 2 pairs of primers were designed and synthesized. Then the 3' and 5'ends of the EST of the SjSDISP from adult Schistosoma japonicum cDNA library were amplified by anchored PCR. After sequencing, a full-length cDNA sequence of the SjSDISP was obtained, and then it was cloned into prokaryotic expression vector pGEX-4T-1. Identified by agarosed gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for the expression under the temperature-dependent condition and Western blot analysis.@*RESULTS@#A 1,071 bp sequence was obtained. Sequence analysis showed that the fragment contained a complete open reading frame (ORF), encoding 278 amino acid residues. This target fragment was cloned into the prokaryotic expression vector pGEX-4T-1, and expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of the expressed fusion recombinant product was 56 kD. Western blot showed that the recombinant protein was recognized by polyclonal rabbit antiserum immunized with Schistosoma japonicum adult worm antigen.@*CONCLUSION@#Cloning of the full-length gene encoding SjSDISP and its bacterial expression were successfully done.
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Asunto principal:
Schistosoma japonicum
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Succinato Deshidrogenasa
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Proteínas Recombinantes
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Datos de Secuencia Molecular
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Secuencia de Bases
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Proteínas del Helminto
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Secuencia de Aminoácidos
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Clonación Molecular
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Homología de Secuencia
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Escherichia coli
Límite:
Animals
Idioma:
Zh
Revista:
Journal of Central South University(Medical Sciences)
Año:
2006
Tipo del documento:
Article