کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2021481 1542352 2008 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Frameshift events associated with the lysyl-tRNA and the rare arginine codon, AGA, in Escherichia coli: A case study involving the human Relaxin 2 protein
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Frameshift events associated with the lysyl-tRNA and the rare arginine codon, AGA, in Escherichia coli: A case study involving the human Relaxin 2 protein
چکیده انگلیسی

Human Relaxin 2 is an insulin-related peptide hormone with a mass of 19,084 Da. The mRNA contains a number of arginine codons that are rarely used by Escherichia coli to produce highly expressed proteins. As a result, expressing this recombinant protein in E. coli is problematic. When human Relaxin 2 was expressed in E. coli BL21 (DE3), several forms of the protein were made. One species had the expected molecular weight (19,084 Da). A second species observed had a molecular weight of 21,244 Da. A third minor species had a molecular weight of 17,118 Da. These aberrant molecular weights can be explained as follows. First, a sequence CGA-AAA-AAG-AGA, containing the rare arginine codons CGA and AGA was the site of the +1 frameshift that generated the 21,244 Da species. Since there was a limited supply of this arginyl-tRNA, the peptidyl-tRNA moved +1 nucleotide to occupy the codon and resumed protein synthesis. Second, a −1 frameshift associated with ‘slippery A’ sequence XXA-AAA-AAG accounted for 10% of the product with a mass of 17,118 Da. Presumably, the shift to −1 also occurred because there was a paucity of the arginyl-tRNAArgucu. Introduction of a plasmid coding for the cognate tRNA for AGA and site directed mutagenesis prevented the formation of both frameshift species.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 60, Issue 2, August 2008, Pages 110–116
نویسندگان
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