Project description:To address the delivery barriers of macromolecular therapeutics, we sought to develop high-throughput peptide barcoding platform as a homogeneous method for macromolecular screening. Implemented in open-source algorithms, BarcodeBabel and PeptideBabel, this approach permits the design of libraries of peptide barcodes and novel penetration sequences optimized for analysis by quantitative mass spectrometry.
Project description:Species identification of fragmentary bones remains a challenging task in archeology and forensics. A species identification method for such fragmentary bones that has recently attracted interest is the use of bone collagen proteins. We developed a method similar to DNA barcoding that reads collagen protein sequences in bone and automatically determines the species by performing sequence database searches. We tested our method using bone samples from 30 vertebrate species ranging from mammals to fish.
Project description:We describe the development of a high-sensitivity protein quantification system called HaloTag protein barcoding assay. The assay involves target protein linking to a unique molecule-counting oligonucleotide by click chemistry.
Project description:Detecting strain-specific barcodes with mass spectrometry can facilitate the screening of genetically engineered bacterial libraries. Here, we introduce intact protein barcoding, a method to measure protein-based library barcodes and metabolites using flow-injection mass spectrometry (FI-MS). Protein barcodes are based on ubiquitin with N-terminal tags of six amino acids. We demonstrate that FI-MS detects intact ubiquitin proteins and identifies the mass of N-terminal barcodes. In the same analysis, we measured relative concentrations of primary metabolites. We constructed 6 ubiquitin-barcoded CRISPRi strains targeting metabolic enzymes, and analyzed their metabolic profiles and ubiquitin barcodes. FI-MS detected barcodes and distinct metabolome changes in CRISPRi-targeted pathways. We demonstrate the scalability of intact protein barcoding by measuring 132 ubiquitin barcodes in microtiter plates. These results show that intact protein barcoding enables fast and simultaneous detection of library barcodes and intracellular metabolites, opening up new possibilities for mass spectrometry-based barcoding.
Project description:Investigation of transcriptome dynamics of Japanese cedar (Cryptomeria japonica) in winter (Dec. 22-23, 2011) and summer (July 30-31, 2012). We investigated seasonal and diurnal transcriptome dynamics of Japanese cedar (Cryptomeria japonica) by analyzing shoot samples collected at four-hour interval for two days in winter and summer, respectively. We first collected sequence data of expressed genes from shoots to designed microarray probes. Microarray analysis revealed the significant difference of transcripts between summer and winter, and the diurnal transcriptome dynamic in summer.Statistical analysis indicated that about 7.7 % of unique genes showed diurnal rhythms with more than two-fold of peak-to-trough amplitude in summer.