Work

Genetic Incorporation of Unnatural Amino Acids into Proteins in Yeast

Year: 2011

Type: book-chapter

Abstract: Unnatural amino acids can be genetically incorporated into proteins in live cells by using an orthogonal tRNA/aminoacyl-tRNA synthetase pair. Here we describe a method to efficiently express the orthogonal tRNA and synthetase in Saccharomyces cerevisiae, which enables unnatural amino acids to be genetically incorporated into target proteins in yeast with high efficiency. We also describe the use of a yeast strain deficient in the nonsense-mediated mRNA decay, which further increases the unnatural amino acid incorporation efficiency when a stop codon is used to encode the unnatural amino acid. These strategies will facilitate the investigation of proteins and their related biological processes in yeast by exploiting the novel properties afforded by unnatural amino acids.

Source: Methods in molecular biology

DOI: 10.1007/978-1-61779-331-8_12

Authors: Qian Wang, Lei Wang

Institutions: Chinese Academy of Sciences, Tianjin Institute of Industrial Biotechnology, Salk Institute for Biological Studies

Language: English

Topic: RNA and protein synthesis mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funder: National Institutes of Health

Award: Genetically Encoding Novel Amino Acids to Investigate Wnt Signaling in C. elegans