日本血吸虫p50亲免素基因克隆、表达及纯化

李孜; 余新炳; 吴忠道; 徐劲; 胡旭初

doi:10.11847/zgggws2005-21-11-14

日本血吸虫p50亲免素基因克隆、表达及纯化

Clone and expression of schistosoma japonicum p50 immunophilin gene and purification of p50 protein

摘要

摘要:
目的扩增、原核克隆和表达日本血吸虫(Sj)p50亲免素基因全长cDNA, 并进行表达产物的纯化.
方法从成虫RNA中RT-PCR扩增Sjp50亲免素基因完整的编码阅读框后, 将其克隆入pET 30 a(+)原核表达载体中, 异丙基硫代-β-D半乳糖苷(IPTG)诱导高表达后用亲和层析柱对表达产物进行纯化.
结果将Sjp50亲免素基因成功克隆入pET 30 a(+)表达载体中; 诱导表达并成功纯化出重组Sjp50亲免素蛋白.
结论成功克隆Sjp50亲免素基因, 并表达和纯化得到了p50亲免素蛋白, 为研究Sjp50亲免素的功能及其进行疫苗等研究奠定了基础.

Abstract:
Objective To clone and express Schistosoma japonicum(Sj)p 50 immunophilin gene, and to purify its protein.
Methods To amplify p 50 immunophilincs complete coding sequence(CDS)from adultworm mRNA by RT-PCR.cDNA of p 50 immunophilin gene was cloned into the pET30 a(+)vector.The recombinantplasmid was transformed into E.coli BL 21(DE3)and expressed, then the expressed products in E.coli were purified by affinity chromatography column.
Results p 50 immunophilin gene cDNA sequence was obtained and then successfully cloned into pET30 a(+)vector.p 50 immunophilin fusion protein was expressed afterinduced by IPTG and purified with(His)6 atthe Nterminus.
Conclusion Sjp 50 immunophilin gene was successfully cloned, expressed and fusion pr otein was purified, which gave the basis for further studyespecially as a vaccine.

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