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Fusion protein systems designed to increase soluble cytoplasmic expression of heterologous proteins in esherichia coli

机译:设计用于增加大肠杆菌中异源蛋白的可溶性胞质表达的融合蛋白系统

摘要

A fusion sequence having a carrier protein which is preferably an E. coli protein having a predicted solubility probability of at least 90% fused to a target heterologous peptide or protein, and a host cell (especially E. coli) transformed with, or having integrated into its genome, a DNA sequence comprising a DNA encoding a carrier protein as defined herein fused to the DNA sequence of a selected heterologous peptide or protein. This fusion sequence is under the control of an expression control sequence capable of directing the expression of a fusion protein in the cell. An objective of the present invention is to improve the purification process of recombinant fusion proteins by avoiding the initial expression of these fusion proteins in E. coli as insoluble inclusion bodies. The methods and compositions of the present invention permit the production of large amounts of heterologous peptides or proteins in a stable, soluble form in certain host cells which normally express limited amounts of such soluble peptides or proteins. The present invention produces fusion proteins which retain the desirable characteristics of a carrier protein (i.e., stability, solubility, and a high level of expression).
机译:融合序列,其具有载体蛋白和宿主细胞(尤其是大肠杆菌),所述载体蛋白优选为与目标异源肽或蛋白融合的预计溶解度至少为90%的大肠杆菌蛋白,并且宿主细胞(尤其是大肠杆菌)转化或整合了进入其基因组的DNA序列包含与选择的异源肽或蛋白质的DNA序列融合的编码本文定义的载体蛋白的DNA。该融合序列在能够指导融合蛋白在细胞中表达的表达控制序列的控制下。本发明的目的是通过避免这些融合蛋白在大肠杆菌中作为不溶性包涵体的初始表达来改善重组融合蛋白的纯化过程。本发明的方法和组合物允许在某些通常表达有限量的此类可溶性肽或蛋白质的宿主细胞中以稳定,可溶的形式产生大量的异源肽或蛋白质。本发明产生融合蛋白,该融合蛋白保留了载体蛋白的所需特性(即,稳定性,溶解性和高表达水平)。

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