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编码人工蛋白质的合成基因的构建、克隆以及获得融合蛋白的表达研究。

The construction and cloning of synthetic genes coding for artificial proteins and expression studies to obtain fusion proteins.

作者信息

Biernat J, Hasselmann H, Hofer B, Kennedy N, Köster H

机构信息

BIOSYNTECH Biochemische Synthesetechnik GmbH, Hamburg, FRG.

出版信息

Protein Eng. 1987 Aug-Sep;1(4):345-51. doi: 10.1093/protein/1.4.345.

Abstract

Synthetic genes coding for artificial proteins with predefined and nutritionally valuable amino acid compositions have been constructed and cloned in bacterial plasmid vector pKK233-2. The genes were constructed from three easily interchangeable 'cassettes' encoding either essential, non-essential or branched-chain amino acid residues. A potential hairpin loop structure in the mRNA around the region of the ribosome binding site was probably the reason for blockage of translation from this vector. Two selected genes, AHB (containing one copy of each cassette) and A6 (consisting of six copies concatemerized A cassette) were cloned into pUR300, a beta-Gal fusion vector and expressed as fusion proteins beta-Gal-AHB and beta-Gal-A6.

摘要

已构建了编码具有预定且具有营养价值的氨基酸组成的人工蛋白质的合成基因,并将其克隆到细菌质粒载体pKK233-2中。这些基因由三个易于互换的“盒式结构”构建而成,分别编码必需氨基酸、非必需氨基酸或支链氨基酸残基。核糖体结合位点区域周围的mRNA中潜在的发夹环结构可能是该载体翻译受阻的原因。将两个选定的基因AHB(每个盒式结构包含一个拷贝)和A6(由六个串联的A盒式结构组成)克隆到β-半乳糖苷酶融合载体pUR300中,并表达为融合蛋白β-半乳糖苷酶-AHB和β-半乳糖苷酶-A6。

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