RESUMEN
Antarctic krill (Euphausia superba) is Earth's most abundant wild animal, and its enormous biomass is vital to the Southern Ocean ecosystem. Here, we report a 48.01-Gb chromosome-level Antarctic krill genome, whose large genome size appears to have resulted from inter-genic transposable element expansions. Our assembly reveals the molecular architecture of the Antarctic krill circadian clock and uncovers expanded gene families associated with molting and energy metabolism, providing insights into adaptations to the cold and highly seasonal Antarctic environment. Population-level genome re-sequencing from four geographical sites around the Antarctic continent reveals no clear population structure but highlights natural selection associated with environmental variables. An apparent drastic reduction in krill population size 10 mya and a subsequent rebound 100 thousand years ago coincides with climate change events. Our findings uncover the genomic basis of Antarctic krill adaptations to the Southern Ocean and provide valuable resources for future Antarctic research.
Asunto(s)
Euphausiacea , Genoma , Animales , Relojes Circadianos/genética , Ecosistema , Euphausiacea/genética , Euphausiacea/fisiología , Genómica , Análisis de Secuencia de ADN , Elementos Transponibles de ADN , Evolución Biológica , Adaptación FisiológicaRESUMEN
The performance of a hemispherical resonant gyroscope (HRG) is directly affected by the sphericity error of the thin-walled spherical shell of the hemispherical shell resonator (HSR). In the production process of the HSRs, high-speed, high-accuracy, and high-robustness requirements are necessary for evaluating sphericity errors. We designed a sphericity error evaluation method based on the minimum zone criterion with an adaptive number of subpopulations. The method utilizes the global optimal solution and the subpopulations' optimal solution to guide the search, initializes the subpopulations through clustering, and dynamically eliminates inferior subpopulations. Simulation experiments demonstrate that the algorithm exhibits excellent evaluation accuracy when processing simulation datasets with different sphericity errors, radii, and numbers of sampling points. The uncertainty of the results reached the order of 10-9 mm. When processing up to 6000 simulation datasets, the algorithm's solution deviation from the ideal sphericity error remained around -3 × 10-9 mm. And the sphericity error evaluation was completed within 1 s on average. Additionally, comparison experiments further confirmed the evaluation accuracy of the algorithm. In the HSR sample measurement experiments, our algorithm improved the sphericity error assessment accuracy of the HSR's inner and outer contour sampling datasets by 17% and 4%, compared with the results given by the coordinate measuring machine. The experiment results demonstrated that the algorithm meets the requirements of sphericity error assessment in the manufacturing process of the HSRs and has the potential to be widely used in the future.
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DNA methylation is a key epigenetic mechanism orchestrating gene expression networks in many biological processes. Nonetheless, studying the role of specific gene methylation events in fish faces challenges. In this study, we validate the regulation of DNA methylation on empty spiracles homeobox 2 (emx2) expression with decitabine treatment in Chinese tongue sole testis cells. We used the emx2 gene as the target gene and developed a new DNA methylation editing system by fusing dnmt3a with catalytic dead Cas9 (dCas9) and demonstrated its ability for sequence-specific DNA methylation editing. Results revealed that utilizing dCas9-dnmt3a to target emx2 promoter region led to increased DNA methylation levels and decreased emx2 expression in Chinese tongue sole testis cells. More importantly, the DNA methylation editing significantly suppressed the expression of MYC proto-oncogene, bHLH transcription factor (myc), one target gene of emx2. Furthermore, we assessed the off-target effects of dCas9-dnmt3a and confirmed no significant impact on the predicted off-target gene expression. Taken together, we developed the first DNA methylation editing system in marine species and demonstrated its effective editing ability in Chinese tongue sole cells. This provides a new strategy for both epigenetic research and molecular breeding of marine species.
Asunto(s)
Metilación de ADN , Edición Génica , Proteínas de Homeodominio , Testículo , Animales , Masculino , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Testículo/metabolismo , Edición Génica/métodos , Sistemas CRISPR-Cas , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Peces Planos/genética , Regiones Promotoras Genéticas/genética , Proteínas de Peces/genética , Proteínas de Peces/metabolismo , ADN Metiltransferasa 3ARESUMEN
Exosomes are endosome-derived extracellular vesicles about 100 nm in diameter. They are emerging as promising delivery platforms due to their advantages in biocompatibility and engineerability. However, research into and applications for engineered exosomes are still limited to a few areas of medicine in mammals. Here, we expanded the scope of their applications to sex-determining gene studies in early vertebrates. An integrated strategy for constructing the exosome-based delivery system was developed for efficient regulation of dmrt1, which is one of the most widely used sex-determining genes in metazoans. By combining classical methods in molecular biology and the latest technology in bioinformatics, isomiR-124a was identified as a dmrt1 inhibitor and was loaded into exosomes and a testis-targeting peptide was used to modify exosomal surface for efficient delivery. Results showed that isomiR-124a was efficiently delivered to the testes by engineered exosomes and revealed that dmrt1 played important roles in maintaining the regular structure and function of testis in juvenile fish. This is the first de novo development of an exosome-based delivery system applied in the study of sex-determining gene, which indicates an attractive prospect for the future applications of engineered exosomes in exploring more extensive biological conundrums.
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Exosomas , Vesículas Extracelulares , Masculino , Animales , Exosomas/genética , Testículo , Endosomas , Péptidos , MamíferosRESUMEN
The novel cellobiohydrolase gene ctcel7 was identified from Chaetomium thermophilum, and its recombinant protein CtCel7, a member of glycoside hydrolase family 7, was heterologously expressed in Pichia pastoris and biochemically characterized. Compared with commercial hydrolases, purified CtCel7 exhibited superior bifunctional cellobiohydrolase and xylanase activities against microcrystalline cellulose and xylan, respectively, under optimal conditions of 60°C and pH 4.0. Moreover, CtCel7 displayed remarkable thermostability with over 90% residual activity after heat (60°C) treatment for 180 min. CtCel7 was insensitive to most detected cations and reagents and preferentially cleaved the ß-1,4-glycosidic bond to generate oligosaccharides through the continuous saccharification of lignocellulosic substrates, which are crucial for various practical applications. Notably, the hydrolysis effect of a commercial cellulase cocktail on pretreated wheat straw was substantively improved by its combination with CtCel7. Taken together, these excellent properties distinguish CtCel7 as a robust candidate for the biotechnological production of biofuels and biobased chemicals.
RESUMEN
BACKGROUND: Endoglucanase has been extensively employed in industrial processes as a key biocatalyst for lignocellulosic biomass degradation. Thermostable endoglucanases with high catalytic activity at elevated temperatures are preferred in industrial use. To improve the activity and thermostability, site-directed mutagenesis was conducted to modify the N-glycosylation sites of the thermostable ß-1,4-endoglucanase CTendo45 from Chaetomium thermophilum. RESULTS: In this study, structure-based rational design was performed based on the modification of N-glycosylation sites in CTendo45. Eight single mutants and one double mutant were constructed and successfully expressed in Pichia pastoris. When the unique N-glycosylation site of N88 was eliminated, a T90A variant was active, and its specific activity towards CMC-Na and ß-d-glucan was increased 1.85- and 1.64-fold, respectively. The mutant R67S with an additional N-glycosylation site of N65 showed a distinct enhancement in catalytic efficiency. Moreover, T90A and R67S were endowed with extraordinary heat endurance after 200 min of incubation at different temperatures ranging from 30 to 90 °C. Likewise, the half-lives (t 1/2) indicated that T90A and R67S exhibited improved enzyme thermostability at 80 °C and 90 °C. Notably, the double-mutant T90A/R67S possessed better hydrolysis activity and thermal stability than its single-mutant counterparts and the wild type. CONCLUSIONS: This study provides initial insight into the biochemical function of N-glycosylation in thermostable endoglucanases. Moreover, the design approach to the optimization of N-glycosylation sites presents an effective and feasible strategy to improve enzymatic activity and thermostability.