Many natural proteins have been developed into drugs and produced for direct application. Identifying improved hosts to achieve
high-level heterologous protein production is a challenge in the study of heterologous protein expression in recombinant yeast.
In this study, a novel high-throughput assay to screen such overproducing Saccharomyces cerevisiae strains was systematically developed. The protocol designed was based on screening host strain derivatives with increased
superoxide dismutase dependent resistance to oxidative stress. Yeast cells transformed with recombinant plasmid carrying SOD1 gene as a reporter responded exquisitely to oxidative stress induced by elevated concentrations of paraquat. Improved yeast
strains resulting from screening clones subjected to genome shuffling through selective pressure argue for a more effective
screening system compared with traditonal selection. Moreover, this approach can be employed in general biochemical analysis
without utilization of flow cytometry or well plate reader. Therefore, it is expected that the high-throughput assay would
make superior strains producing heterologous proteins. 相似文献
Research has revealed that most chlorophyllous explants/plants in vitro have the ability to grow photoautotrophically (without
sugar in the culture medium), and that the low or negative net photosynthetic rate of plants in vitro is not due to poor photosynthetic
ability, but to the low CO2 concentration in the air-tight culture vessel during the photoperiod. Moreover, numerous studies have been conducted on improving
the in vitro environment and investigating its effects on growth and development of cultures/plantlets on nearly 50 species
since the concept of photoautotrophic micropropagation was developed more than two decades ago. These studies indicate that
the photoautotrophic growth in vitro of many plant species can be significantly promoted by increasing the CO2 concentration and light intensity in the vessel, by decreasing the relative humidity in the vessel, and by using a fibrous
or porous supporting material with high air porosity instead of gelling agents such as agar. This paper reviews the development
and characteristics of photoautotrophic micropropagation systems and the effects of environmental conditions on the growth
and development of the plantlets. The commercial applications and the perspective of photoautotrophic micropropagation systems
are discussed. 相似文献
Rift Valley Fever virus (RVFV) is an enzootic virus that causes extensive morbidity and mortality in domestic ruminants in Africa, and it has shown the potential to invade other areas such as the Arabian Peninsula. Here, we develop methods for linking mathematical models to real-world data that could be used for continent-scale risk assessment given adequate data on local host and vector populations. We have applied the methods to a well-studied agricultural region of California with 1 million dairy cattle, abundant and competent mosquito vectors, and a permissive climate that has enabled consistent transmission of West Nile virus and historically other arboviruses. Our results suggest that RVFV outbreaks could occur from February–November, but would progress slowly during winter–early spring or early fall and be limited spatially to areas with early increases in vector abundance. Risk was greatest in summer, when the areas at risk broadened to include most of the dairy farms in the study region, indicating the potential for considerable economic losses if an introduction were to occur. To assess the threat that RVFV poses to North America, including what-if scenarios for introduction and control strategies, models such as this one should be an integral part of the process; however, modeling must be paralleled by efforts to address the numerous remaining gaps in data and knowledge for this system. 相似文献
Tunable triple-peaks with the transmission intensity of more than 90% plasmonically induced transparency metamaterial resonator based on nested double π-shaped metallic structure is proposed at the terahertz frequency region, which is consisted of three sets of gold nanorods with different sizes placed on a dielectric substrate of SiO2. The coupling effect of localized electric field between different parts of the proposed structure can be used to explain the physical mechanism of three transparent windows. The finite-difference time-domain (FDTD) is used to study the spectral properties of the proposed structure, and the influence of the size of the nanorods and the relative distance between them on the spectral characteristics are also discussed. It can be seen that some obvious shift phenomena occur in the spectra with the change of these nanorods. These results indicate that the proposed structure opens up new avenues in many related applications, especially for multi-channel filters, optical switches, and sensors.
Untargeted metabolomics intends to objectively analyze a wide variety of compounds. Their diverse physicochemical properties make it difficult to choose an appropriate reconstitution solvent after sample evaporation without influencing the chromatography or hamper column sorbent integrity.
Objectives
The study aimed to identify the most appropriate reconstitution solvent for blood plasma samples in terms of feature recovery, four endogenous compounds, and one selected internal standard.
Methods
We investigated several reconstitution solvent mixtures containing acetonitrile and methanol to resolve human plasma extract and evaluated them concerning the peak areas of tryptophan-d5, glucose, creatinine, palmitic acid, and the phophatidylcholine PC(P-16:0/P-16:0), as well as the total feature count
Results
Results indicated that acetonitrile containing 30% methanol was best suited to match all tested criteria at least for human blood plasma samples.
Conclusion
Despite identifying the mixture of acetonitrile and methanol being suitable as solvent for human blood plasma extracts, we recommend to systematically test for an appropriate reconstitution solvent for each analyzed biomatrix.
Hepatocellular carcinoma (HCC), with life‐threatening malignant behaviours, often develops distant metastases and is the fourth most common primary cancer in the world, having taken millions of lives in Asian countries such as China. The novel miR‐3677‐3p is involved in a high‐expression‐related poor prognosis in HCC tissues and cell lines, indicating oncogenesis functions in vitro and in vivo. Initially, we confirmed the inhibition of proliferation, migration and invasion in miR‐3677‐3p knock‐down MHCC‐97H and SMMC‐7721 cell lines, which are well known for their high degree of invasiveness. Then, we reversed the functional experiments in the low‐miR‐3677‐3p‐expression Hep3B cell line via overexpressing miR‐3677‐3p. In nude mice xenograft and lung metastasis assays, we found suppressor behaviours, smaller nodules and low density of organ spread, after injection of cells transfected with shRNA‐miR‐3677‐3p. A combination of databases (Starbase, TargetScan and MiRgator) illustrated miR‐3677‐3p targets, and it was shown to suppress the expression of SIRT5 in a dual‐luciferase reporter system. To clarify the conclusions of previous ambiguous research, we up‐regulated SIRT5 in Hep3B cells, and rescue tests were established for confirmation that miR‐3677‐3p suppresses SIRT5 to enhance the migration and invasion of HCC. Interestingly, we discovered hypoxia‐induced miR‐3677‐3p up‐regulation benefited HCC malignancy and invasiveness. In conclusion, the overexpression of miR‐3677‐3p mediated SIRT5 inhibition, which could increase proliferation, migration and invasion of HCC in hypoxic microenvironments. 相似文献