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首页> 外文期刊>Asian Pacific Journal of Cancer Prevention >Preliminary Research on the Expression, Purification and Function of the Apoptotic Fusion Protein, Sival
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Preliminary Research on the Expression, Purification and Function of the Apoptotic Fusion Protein, Sival

机译:较初步研究凋亡融合蛋白的表达,纯化和功能,雄鹿

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The objective of the present study was to investigate cloning, expression, and functions of the recombinant protein, Siva1. Siva1 gene was synthesized by RT-PCR from HCT116 cells. Plasmids were cleaved with the restriction endonuclease, BamH1/Sal1 and products were connected to pQE30, which underwent cleavage by BamH1/Sal1. The recombinant plasmid, pQE30-Siva1, was identified after digestion with restriction endonucleases followed by transformation into E. coli M15. Expression of Siva1 was induced by IPTG and identified by SDS-PAGE following purification with affinity chromatography. The results showed that size of Siva1 was 12 kDa, consistent with the molecular weight of the His-Siva1 fusion protein. Functional test demonstrated that Siva1 significantly inhibited the invasion and migration of HCT116 cells. It may thus find clinical application for control of cancers.
机译:本研究的目的是研究重组蛋白Siva1的克隆,表达和功能。通过RT-PCR从HCT116细胞中合成SiVA1基因。质粒用限制性内切核酸酶切割,BamH1 / Sal1和产物连接到PQE30,由BamH1 / Sal1接受切割。在用限制性内切核酸酶的消化后鉴定重组质粒pQe30-Siva1,然后通过转化为大肠杆菌M15。通过IPTG诱导SiVA1的表达,并通过亲和层析纯化后通过SDS-PAGE鉴定。结果表明,SiVA1的尺寸为12kDa,与His-Siva1融合蛋白的分子量一致。功能试验证明SiVA1显着抑制HCT116细胞的侵袭和迁移。因此,它可以找到对癌症的临床应用。

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