After the PACAPintein-CBD fusion protein was purified by chitin-affinity chromatography, the self-cleavage activity of the intein was induced by DTT and the rhPACAP was released from the chitin-bound intein tag. The activity of the rhPACAP to stimulate cyclic AMP accumulation was detected using ...
coli 中表現得到可溶的蛋白,於是將 ORF151 穿膜區域去除形成截短的 ORF151,並於 N 端接有 CBD-Ssp intein 序列 (命名為 intein-ORF151ΔTMD),並將 ORF106 之 lipobox 去除,形成截短的 ORF106 (命名為 ORF106ΔLpp) 並進行表現.以共同... 侯萱吾,H. Hou 被引量: 0发表: 2014年 基于内含肽介导...
然后将 p MYB12 9中 intein- CBD基因片段移插入上述经改造的p Ex Sec I中 ,构建成含内蛋白子的表达载体 p Ex IC。2) intein 蛋白质内含子 1. Use of Ssp dnaB Mini-intein as a Fusion Partner for Preparation of Recombinant Human Brain Natriuretic Peptide; Ssp dnaB蛋白质内含子介导的重组人脑...
However hydrophobicity analysis of the N-terminal regions of Ssp DnaB intein showed hydrophilic properties, therefore we purposely kept the CBD domain appearing before the Ssp DnaB intein (as derived from commercial pTWIN1 vector) to act as a hydrophobic linker between PHA synthase and Ssp DnaB...
intein tag consisting of the intein and CBD. The CBD is inserted in a loop region of the engineered Sce VMA intein such that the cleavage activity of the intein is not affected. The engineered intein can undergo cleavage at its C-terminus triggered by thiol-induced cleavage at its ...
Many of these methods have the drawbacks of either leaving the recombinant tag attached to the protein or requiring the addition of a protease which then must be removed by further chromatographic steps. The fusion of a self-cleaving intein sequence followed by a chitin-binding domain (CBD) ...
The expression of IFN beta fused with intein tag was confirmed by western blot using anti-CBD. In addition, the soluble form of IFN beta fusion protein was determined to be about 35%, using the SDS-PAGE technique. In conclusion, the expression of IFN beta fused wit...
The expression of IFNβ fused with intein tag was confirmed by western blot using anti-CBD. In addition, the soluble form of IFNβ fusion protein was determined to be about 35%, using the SDS-PAGE technique. In conclusion, the expression of IFNβ fused with INT and CBD can be a ...
Peptide epitopes possessing an N-terminal cysteine were ligated to the chitin bound CBD tag. We demonstrate that the resulting peptide columns permit the highly specific and efficient affinity purification of antibodies from animal sera.doi:10.1016/j.jim.2003.07.005Luo Sun...
In this study, the prokaryotic expression plasmid pTwin1-N-IFN-gamma was constructed to express Cyprinus carpio (common carp) IFN-gamma fused with a chitin binding domain (CBD) and a self-cleavable intein-tag, Synechocystis sp DnaB. The recombinant protein CBD-DnaB-IFN-gamma with the ...