[干扰素诱导跨膜蛋白1基因的PCR扩增、克隆及蛋白表达]。

Yu-hu Liu, Dong Zhong, Juan Liu, Zhen-shu Zhang, Cun-long Chen, Jin-bao Wu, Bing Xiao, Wen-ying Guo
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引用次数: 0

摘要

目的:研究干扰素诱导跨膜蛋白-1 (IFITMP-1)基因的PCR扩增、克隆及蛋白表达。方法:以IFITMP-1基因cDNA片段为模板,采用Pfu酶PCR扩增IFITMP-1基因。经EcoRI和HindIII酶切后,将目的基因片段与pUCm-T质粒连接并测序。将IFITMP-1基因克隆到pET-Trx蛋白表达质粒中,优化蛋白表达条件。结果:IFITMP-1基因cDNA片段的PCR产物长度约为1000bp。将IFITMP-1基因按正确序列成功插入pUCm-T质粒中,实现了将IFITMP-1基因克隆到pET-Trx蛋白表达质粒中。IPTG诱导后检测pUCm-T质粒与IFITMP-1基因融合蛋白的表达。结论:成功扩增、克隆IFITMP-1基因及其蛋白表达,为进一步研究IFITMP-1基因在结直肠癌中的作用提供了基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[PCR amplification, cloning and protein expression of interferon-inducible transmembrane protein-1 gene].

Objective: To study the PCR amplification, cloning and protein expression of interferon-inducible transmembrane protein-1 (IFITMP-1) gene.

Methods: With the cDNA fragment containing IFITMP-1 gene as template, IFITMP-1 gene was amplified using Pfu enzyme by means of PCR. After EcoRI and HindIII digestion, the target gene fragment was linked to pUCm-T plasmid and sequenced. The IFITMP-1 gene was cloned into pET-Trx protein expression plasmid, and the condition for protein expression was optimized.

Results: The length of the PCR product of IFITMP-1 gene-containing cDNA fragment was about 1000 bp. The IFITMP-1 gene was successfully inserted into pUCm-T plasmid with correct sequence and cloning of the IFITMP-1 gene into the pET-Trx protein expression plasmid was achieved. Expression of the fusion protein of pUCm-T plasmid and IFITMP-1 gene was detected after IPTG induction.

Conclusion: Successful amplification and cloning of the IFITMP-1 gene and its protein expression may facilitate further study of the role of IFITMP-1 gene in colorectal cancer.

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