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首页> 外文期刊>Protein Expression and Purification >Affinity tag effect on the salt rejection potential of Halomonas elongata aquaporin incorporated in thin film nanocomposite membrane
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Affinity tag effect on the salt rejection potential of Halomonas elongata aquaporin incorporated in thin film nanocomposite membrane

机译:关于卤代薄膜含水金蛋白的盐排斥潜力的亲和标签效应掺入薄膜纳米复合材料膜中

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摘要

In this study, effect of affinity tags, Histidine (His) and Glutathione-S-Transferase (GST), on the activity of halophilic aquaporin was analyzed. The gene coding for H. elongata aquaporin was cloned into pET28a vector and expressed in E. coli. BL21 successfully. Stopped flow light scattering measurements showed that His-tagged aquaporin is functional. The difference in the filtration parameters caused by affinity tags were determined by using thin film composite nano-filtration (NFC) membranes prepared with the aquaporins. At 100 mM salt concentration, water permeability (L/m(2).h) and the % salt rejection of NFC membranes produced with the His-tagged aquaporin was found to be higher than that of the membrane with GST-tagged aquaporin. Salt rejection of His-tagged aquaporin-membrane was found to be 53% with a lower solute permeability value (B). Use of short affinity tag (His tag) for cloning resulted in higher solute rejection ability of TFC membranes prepared with H. elongata aquaporins.
机译:在本研究中,分析了亲和标签,组氨酸(HIS)和谷胱甘肽-S-转移酶(GST)对嗜盐水蛋白的活性的影响。编码H.Elongata Aquaporin的基因编码成PET28a载体并在大肠杆菌中表达。 BL21成功。停止流动光散射测量表明,他标记的水通道蛋白是功能性的。通过使用用水上蛋白制备的薄膜复合纳米过滤(NFC)膜来确定由亲和标签引起的过滤参数的差异。在100mm盐浓度下,发现用HIS标记的水蛋白产生的NFC膜的水渗透率(L / m(2).h)和NFC膜的%盐抑制率高于具有GST标记的水素的膜的NFC膜。发现其标记的水蛋白膜的盐排斥是53%,溶质渗透率较低(B)。使用短亲和标签(他的标签)用于克隆,导致用H.Legata Aquaporins制备的TFC膜的溶质抑制能力较高。

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