A5091134405- Bacteriophages and microbial interactions
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Advanced biosensing and bioanalysis techniques
- Genomics and Phylogenetic Studies
- Monoclonal and Polyclonal Antibodies Research
- Chemical Synthesis and Analysis
- Advanced Biosensing Techniques and Applications
- Plant biochemistry and biosynthesis
- Microbial Natural Products and Biosynthesis
- DNA and Nucleic Acid Chemistry
- Evolution and Genetic Dynamics
- Natural product bioactivities and synthesis
- Microbial Metabolic Engineering and Bioproduction
- Research in Social Sciences
- CRISPR and Genetic Engineering
- Biochemical and Molecular Research
Scripps Research Institute
2016-2021
Novo Nordisk Foundation
2016-2021
Technical University of Denmark
2016-2021
University of California, Davis
2016
How do ageing bacterial colonies generate adaptive mutants? Over a period of two months, we isolated on outgrowing mutants able to use new carbon source, and sequenced their genomes. This allowed us uncover exquisite details the molecular mechanism behind adaptation: most mutations were located in just few hotspots genome, over time, increasingly consistent with involvement 8-oxo-guanosine, formed exclusively transcribed strand. work provides strong support for retromutagenesis as general...
Previously, we reported the creation of a semi-synthetic organism (SSO) that stores and retrieves increased information by virtue stably maintaining an unnatural base pair (UBP) in its DNA, transcribing corresponding nucleotides into codons anticodons mRNAs tRNAs, then using them to produce proteins containing noncanonical amino acids (ncAAs). Here report systematic extension effort optimize SSO exploring variety deoxy- ribonucleotide analogues. Importantly, this includes first vivo...
Nucleoside triphosphates play a central role in biology, but efforts to study these roles have proven difficult because the levels of are tightly regulated cell and individual can be label or modify. In addition, many synthetic biology focused on development unnatural nucleoside that perform specific functions cellular environment. general, both would facilitated by general means directly introduce desired into cells. Previously, we demonstrated recombinant expression triphosphate...
We have developed a family of unnatural base pairs (UBPs), exemplified by the pair formed between dNaM and dTPT3, for which pairing is mediated not complementary hydrogen bonding but hydrophobic packing forces. These UBPs enabled creation first semisynthetic organisms (SSOs) that store increased genetic information use it to produce proteins containing noncanonical amino acids. However, retention was poor in some sequence contexts. Here, optimize SSO, we synthesize two novel...
The evolution of microorganisms often involves changes unclear relevance, such as transient phenotypes and sequential development multiple adaptive mutations in hotspot genes. Previously, we showed that ageing colonies an E. coli mutant unable to produce cAMP when grown on maltose, accumulated the crp gene (encoding a global transcription factor) genes involved pyrimidine metabolism cmk; combined both cmk enabled fermentation maltose (which usually requires cAMP-mediated Crp activation for...
Bacterial surface display is an attractive technique for the production of cell-anchored, functional proteins and engineering whole-cell catalysts. Although various outer membrane have been used display, easy versatile high-throughput-compatible assay evaluating developing systems missing. Using a single domain antibody (also called nanobody) with high affinity green fluorescent protein (GFP), we constructed system that allows fast, fluorescence-based detection displayed proteins. The hybrid...
We have developed semisynthetic organisms (SSOs) that by virtue of a family synthetic, unnatural base pairs (UBPs), store and retrieve increased information. To date, transcription in the SSOs has relied on heterologous expression RNA polymerase from T7 bacteriophage; here, we explore placing under control endogenous host multisubunit polymerase. The results demonstrate E. coli is able to transcribe DNA containing UBP with most optimal identified date it should be possible select for uptake...
Diterpenes form a vast and diverse class of natural products both ecological economic importance. Class II diterpene synthase (diTPS) enzymes control the committed biosynthetic reactions underlying chemical diversity. Homology modelling with site-directed mutagenesis identified two active-site residues in horehound (Marrubium vulgare) diTPS peregrinol diphosphate (MvCPS1); residue substitutions abolished unique MvCPS1-catalysed water-capture reaction at C9 redirected enzyme activity toward...