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Genetic code expansion, protein expression and protein functionalisation in Bacillus subtilis.

07:00 EST 13th February 2020 | BioPortfolio

Summary of "Genetic code expansion, protein expression and protein functionalisation in Bacillus subtilis."

The site-specific chemical modification of proteins through incorporation of noncanonical amino acids enables diverse applications, such as imaging, probing and expanding protein functions, as well as to precisely engineer therapeutics. Here we report a general strategy that allows the incorporation of noncanonical amino acids into target proteins using the amber suppression method and their efficient secretion in the high potential microbial production system Bacillus subtilis. This facilitates efficient purification of target proteins directly from the supernatant, followed by their functionalisation using click chemistry. We used this strategy to introduce norbornene lysine site-specifically into a single chain antibody and functionalise it with fluorophores for the detection of human target proteins.

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This article was published in the following journal.

Name: ACS synthetic biology
ISSN: 2161-5063
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Medical and Biotech [MESH] Definitions

A small maf protein that forms dimers with NRF1 protein; NRF2 PROTEIN; and P45 NF-E2 PROTEIN. MafF complexes bind Maf recognition elements to regulate tissue-specific GENETIC TRANSCRIPTION.

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A TALE-type homeodomain protein and transcription factor that functions as a regulator of PAX6 PROTEIN expression and as an activator of PLATELET FACTOR 4 gene expression. It is essential for hematopoiesis, differentiation of MEGAKARYOCYTES, and vascular patterning. It may also have a role in the induction of myeloid leukemias.

A heterogeneous-nuclear ribonucleoprotein found in the CELL NUCLEUS and the CYTOPLASM. Heterogeneous-nuclear ribonucleoprotein K has been implicated in the regulation of gene expression at nearly all levels: GENETIC TRANSCRIPTION; mRNA processing (RNA PROCESSING, POST-TRANSCRIPTIONAL), mRNA transport, mRNA stability, and translation (TRANSLATION, GENETIC). The hnRNP protein has a strong affinity for polypyrimidine-rich RNA and for single-stranded polypyrimidine-rich DNA. Multiple hnRNP K protein isoforms exist due to alternative splicing and display different nucleic-acid-binding properties.

A DNA-binding protein that represses GENETIC TRANSCRIPTION of target genes by recruiting HISTONE DEACETYLASES. Aberrant Blc-6 expression is associated with certain types of human B-CELL LYMPHOMA.

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