Crustacean vitellogenesis is a process that involves Vitellin, produced via endoproteolysis of its precursor, which is designated as Vitellogenin (Vtg). The Vtg gene, mRNA and protein regulation involve several environmental factors and physiological processes, including gonadal maturation and moult stages, among others. Once the Vtg gene, mRNAs and protein are obtained, it is possible to establish the relationship between the elements that participate in their regulation, which could either be species-specific, or tissue-specific. This work is a systematic analysis that compares the similarities and differences of Vtg genes, mRNA and Vtg between the crustacean species reported in databases with respect to that obtained from the transcriptome of Callinectes arcuatus, C. toxotes, Penaeus stylirostris and P. vannamei obtained with MiSeq sequencing technology from Illumina. Those analyses confirm that the Vtg obtained from selected species will serve to understand the process of vitellogenesis in crustaceans that is important for fisheries and aquaculture. RESUMENLa vitelogénesis de los crustáceos es un proceso que involucra la vitelina, producida a través de la endoproteólisis de su precursor llamado Vitelogenina (Vtg). La regulación del gen Vtg, los ARNm y la Vtg involucra factores ambientales y procesos fisiológicos, incluyendo: maduración gonadal, etapas de muda, entre otros. Con el gen Vtg, los ARNm y la proteína obtenidos, es posible correlacionar los elementos que participan en su regulación, pudiendo ser especie-específicos o tejido-específicos. Este trabajo es un análisis sistemático que compara las similitudes y diferencias
Background: Recent estimates indicate that the COVID-19 pandemic, which is caused by the SARS-CoV-2 virus, could be effectively controlled via the development and implementation of diagnostic tools such as quantitative reverse transcription PCR (RT-qPCR). However, this reaction often generates false-negative results due to novel mutations and can also be affected by the secondary structure of the RNA transcripts that derive from the gene sequence used for diagnostic purposes. Methods: Using high-performance computing, we consolidated a global SARS-CoV-2 genome repository encompassing 19,317 genomes from the GenBank database and 107,259 from the GISAID database to generate monthly SARS-CoV-2 consensus sequences from January to December 2020. Results: These sequences were then used to create ORF8-specific primers and probes to validate single and multiplex RT-qPCR protocols both in silico and experimentally using genes E (Berlin protocol) and N (CDC protocol) as targets. Conclusions: Our findings demonstrated that RT-qPCR Ct values were improved by the inclusion of either a denaturing solution composed of tetraethylammonium chloride (TEA) and dimethyl sulfoxide (DMSO) and by adjusting nucleotide proportions based on the SARS-CoV-2 genome.
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