Modeling of DNA technology of species identification of plant raw materials for brewing

Развитие молекулярно-генетических технологий оценки пивоваренного сырья актуально с позиции их внедрения в систему идентификации и прослеживаемости в контексте расширения оценочных критериев менеджмента качества. Целью настоящей работы являлось моделирование ДНК-технологии видовой идентификации растительного сырья для пивоварения. Подобраны протоколы экстракции нуклеиновых кислот, постановки ПЦР и ПДРФ-анализа с соответствующими комплектами реагентов, направленные на практическое воспроизведение генетического тестирования пробоподготовленного биоматериала. Представлены результаты выравнивания и рестрикционного картирования амплифицируемых нуклеотидных последовательностей локуса хлоропластной ДНК ячменя, пшеницы, ржи, кукурузы, риса и хмеля. Установлено, что наличие видоспецифичных нуклеотидных замен и инделей в анализируемом локусе позволяет идентифицировать растительное сырье для пивоварения методом прямого секвенирования ПЦР-продукта. Последующий совокупный анализ данных in silico моделирования ПЦР-ПДРФ-профилей по трем эндонуклеазам рестрикции подтвердил диагностическую ценность подобранных ферментов. The development of molecular genetic technologies for evaluating brewing raw materials is relevant from the point of view of their introduction into the identification and traceability system in the context of expanding the evaluation criteria of quality management. The purpose of this work was to simulate the DNA technology of species identification of plant raw materials for brewing. Protocols for the extraction of nucleic acids, PCR and RFLP analysis with the corresponding reagent kits were selected, aimed at the practical reproduction of genetic testing of the prepared biomaterial. The results of alignment and restriction mapping of amplified nucleotide sequences of the chloroplast DNA locus of barley, wheat, rye, corn, rice and hops are presented. It was found that the presence of species-specific nucleotide substitutions and indels in the analyzed locus makes it possible to identify plant raw materials for brewing by direct sequencing of the PCR product. Subsequent aggregate analysis of the data in silico modeling of PCR-RFLP profiles for three restriction endonucleases confirmed the diagnostic value of the selected enzymes

If Not Now, When? Nonserotype Pneumococcal Protein Vaccines

The sudden emergence and global spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have greatly accelerated the adoption of novel vaccine strategies, which otherwise would have likely languished for years. In this light, vaccines for certain other pathogens could certainly benefit from reconsideration. One such pathogen is Streptococcus pneumoniae (pneumococcus), an encapsulated bacterium that can express >100 antigenically distinct serotypes. Current pneumococcal vaccines are based exclusively on capsular polysaccharide—either purified alone or conjugated to protein. Since the introduction of conjugate vaccines, the valence of pneumococcal vaccines has steadily increased, as has the associated complexity and cost of production. There are many pneumococcal proteins invariantly expressed across all serotypes, which have been shown to induce robust immune responses in animal models. These proteins could be readily produced using recombinant DNA technology or by mRNA technology currently used in SARS-CoV-2 vaccines. A door may be opening to new opportunities in affordable and broadly protective vaccines.

Development of a Freeze-Dried CRISPR-Cas12 Sensor for Detecting Wolbachia in the Secondary Science Classroom

Training the future synthetic biology workforce requires opportunity and exposure to biotechnology concepts and activities in secondary education. Detecting Wolbachia bacteria in arthropods using PCR has become a common way for secondary students to investigate and apply DNA technology in the science classroom. Despite this framework, cutting-edge biotechnologies like CRISPR-based diagnostics have yet to be widely implemented in the classroom. To address this gap, we present a freeze-dried CRISPR-Cas12 sensing reaction to complement traditional DNA technology education and teach synthetic biology concepts. The reactions accurately detect Wolbachia from arthropod-derived PCR samples in under 2 hours and can be stored at room temperature for over a month without appreciable degradation. The reactions are easy-to-use and cost less than $40 to implement for a classroom of 22 students including the cost of reusable equipment. We see this technology as an accessible way to incorporate synthetic biology education into existing biology curriculum, which will expand biology educational opportunities in science, technology, engineering, and mathematics (STEM) education.