The site-specific nature and ease of handling of clustered regularly interspaced palindromic repeat(CRISPR)/Cas9 has fulfilled the decades-long goal of genome engineering,and thus has pushed the biological sciences into a new era in which site-directed manipulation of DNA is no longer a short slab(Doudna and Charpentier,2014).The incomparable power of this system also enabled unprecedented new opportunities for practical applications in agriculture,ecosystem and human health(Liu,2017).However,with the extremely rapid development and application of CRISPR system,concerns about its bio-safety have been increasing(Caplan et al.,2015;Taning et al.,2017).While multiple studies have addressed this issue,alternative options for regulating Cas9 are of great interest.As has been proved by increasing evidences,another well?known concern is the off-target effects,which could not only confound research experiments or breeding,but also cause unwanted side effects in the forthcoming therapeutic uses(Fu et aL,2013;Taning et al.,2017;Zhang et al.,2018). 相似文献
Upland cotton (Gossypium hirsutum) is the world's largest source of natural fibre and dominates the global textile industry. Hybrid cotton varieties exhibit strong heterosis that confers high fibre yields, yet the genome‐wide effects of artificial selection that have influenced Upland cotton during its breeding history are poorly understood. Here, we resequenced Upland cotton genomes and constructed a variation map of an intact breeding pedigree comprising seven elite and 19 backbone parents. Compared to wild accessions, the 26 pedigree accessions underwent strong artificial selection during domestication that has resulted in reduced genetic diversity but stronger linkage disequilibrium and higher extents of selective sweeps. In contrast to the backbone parents, the elite parents have acquired significantly improved agronomic traits, with an especially pronounced increase in the lint percentage. Notably, identify by descent (IBD) tracking revealed that the elite parents inherited abundant beneficial trait segments and loci from the backbone parents and our combined analyses led to the identification of a core genomic segment which was inherited in the elite lines from the parents Zhong 7263 and Ejing 1 and that was strongly associated with lint percentage. Additionally, SNP correlation analysis of this core segment showed that a non‐synonymous SNP (A‐to‐G) site in a gene encoding the cell wall‐associated receptor‐like kinase 3 (GhWAKL3) protein was highly correlated with increased lint percentage. Our results substantially increase the valuable genomics resources available for future genetic and functional genomics studies of cotton and reveal insights that will facilitate yield increases in the molecular breeding of cotton. 相似文献
Tobacco stalk (TS), a major agricultural waste abundant in pectin, has resulted in concerns about the need for its reuse. The nicotine in TS is considered a chemical that is to\xic and hazardous to the environment.
Results
In this study, Bacillus tequilensis CAS-MEI-2-33 was isolated from cigar wrappers to produce alkaline pectinase using TS. Subsequently, the medium and fermentation conditions for the production of pectinase by B. tequilensis CAS-MEI-2-33 were optimized. The optimal fermentation period, pH of the initial fermentation medium, concentration of TS, and inoculum amount for B. tequilensis CAS-MEI-2-33 were 40 h, 40 g/L, 7.0, and 3%, respectively. Under optimal conditions, the pectinase activity was 1370 U/mL. Then, the enzymatic properties, such as the optimum pH, reaction temperature, temperature stability, and effects of metal ions, were studied. The optimal pH was determined to be 10.0, indicating that the enzyme was an alkaline pectinase. The optimal temperature was 40 °C, and pectinase activity was stable at 40 °C. The Ag+ metal ions were shown to remarkably promote enzyme activity. The pectinase was partly purified by ammonium sulfate precipitation, ion exchange chromatography, and Sephacryl S-100 chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and LC-MS/MS analyses were utilized to analyze the pectinase.
Conclusions
This study provided a new alkaline pectinase candidate and a new strategy for the use of TS.
Rho‐associated coiled‐coil kinase 1 (ROCK1) is proposed to be implicated in Aβ suppression; however, the role for ROCK1 in amyloidogenic metabolism of amyloid precursor protein (APP) to produce Aβ was unknown. In the present study, we showed that ROCK1 kinase activity and its APP binding were enhanced in AD brain, resulting in increased β‐secretase cleavage of APP. Furthermore, we firstly confirmed that APP served as a substrate for ROCK1 and its major phosphorylation site was located at Ser655. The increased level of APP Ser655 phosphorylation was observed in the brain of APP/PS1 mice and AD patients compared to controls. Moreover, blockade of APP Ser655 phosphorylation, or inhibition of ROCK1 activity with either shRNA knockdown or Y‐27632, ameliorated amyloid pathology and improved learning and memory in APP/PS1 mice. These findings suggest that activated ROCK1 targets APP Ser655 phosphorylation, which promotes amyloid processing and pathology. Inhibition of ROCK1 could be a potential therapeutic approach for AD. 相似文献
Aging is an independent risk factor for vascular diseases. Perivascular adipose tissue (PVAT), an active component of the vasculature, contributes to vascular dysfunction during aging. Identification of underlying cell types and their changes during aging may provide meaningful insights regarding the clinical relevance of aging‐related vascular diseases. Here, we take advantage of single‐cell RNA sequence to characterize the resident stromal cells in the PVAT (PVASCs) and identified different clusters between young and aged PVASCs. Bioinformatics analysis revealed decreased endothelial and brown adipogenic differentiation capacities of PVASCs during aging, which contributed to neointimal hyperplasia after perivascular delivery to ligated carotid arteries. Mechanistically, in vitro and in vivo studies both suggested that aging‐induced loss of peroxisome proliferator‐activated receptor‐γ coactivator‐1 α (PGC1α) was a key regulator of decreased brown adipogenic differentiation in senescent PVASCs. We further demonstrated the existence of human PVASCs (hPVASCs) and overexpression of PGC1α improved hPVASC delivery‐induced vascular remodeling. Our finding emphasizes that differentiation capacities of PVASCs alter during aging and loss of PGC1α in aged PVASCs contributes to vascular remodeling via decreased brown adipogenic differentiation. 相似文献
The reduction of nuclear fast red (NFR) stain by sodium tetrahydroboron was catalyzed in the presence of silver ions (Ag+). The fluorescence properties of reduced NFR differed from that of NFR. The product showed fluorescence emission at 480 nm with excitation at 369 nm. Furthermore, the fluorescence intensity of the mixture increased strongly in the presence of Ag+ and Britton–Robinson buffer at pH 4.78. There was a good linear relationship between increased fluorescence intensity (ΔI) and Ag+ concentration in the range 5.0 × 10?9 to 5.0 × 10?8 M. The correlation coefficient was 0.998, and the detection limit (3σ/k) was 1.5 × 10?9 M. The colour of the reaction system changed with variation in Ag+ concentration over a wide range. Based on the colour change, a visual semiquantitative detection method for recognition and sensing of Ag+ was developed for the range 1.0 × 10?8 to 5.0 × 10?4 M, with an indicator that was visible to the naked eye. Therefore, a sensitive, simple method for determination of Ag+ was developed. Optimum conditions for Ag+ detection, the effect of other ions and the analytical application of Ag+ detection of synthesized sample were investigated. 相似文献
Taking advantage of the compelling properties of d ‐penicillamine (d ‐PA) combined with copper, a method for the sensitive and selective determination of d ‐PA was established using copper nanocluster (Cu NC)‐based fluorescence enhancement. d ‐PA molecules containing a thiol compound showed a strong tendency to combine with the surface of Cu NCs, causing the re‐dispersion of nanoclusters and therefore fluorescence intensity was enhanced. Fluorescence enhancement efficiency of Cu NCs induced by d ‐PA was linear, with the d ‐PA concentration varying from 0.6–30 μg ml?1 (R2 = 0.9952) and with a detection limit of 0.54 μg ml?1. d ‐PA content in human urine samples was detected with recoveries of 104.8–112.99%. Fluorescence‐enhanced determination of d ‐PA using Cu NCs was established for the first time and this rapid, easy and sensitive method should attract much attention for this application. 相似文献