为提取高质量的山药总RNA,以铁棍山药茎段为材料,采用Trizol法、RNAiso for Polysacchariderich Plant Tissue法、改良CTAB法和试剂盒法提取总RNA,利用琼脂糖凝胶电泳分析所提取RNA的完整性,并用超微量分光光度计分析RNA的纯度.结果显...为提取高质量的山药总RNA,以铁棍山药茎段为材料,采用Trizol法、RNAiso for Polysacchariderich Plant Tissue法、改良CTAB法和试剂盒法提取总RNA,利用琼脂糖凝胶电泳分析所提取RNA的完整性,并用超微量分光光度计分析RNA的纯度.结果显示采用Trizol法难以提RNA,RNAiso for Polysacchariderich Plant Tissue法提取效果不理想,存在DNA和蛋白质污染,而用改良CTAB法和试剂盒法提取到的总RNA纯度高、完整性好,其中,改良CTAB法较试剂盒法价格便宜,但提取过程较烦琐.因此,应根据实验需要选择CTAB法或试剂盒法提取铁棍山药总RNA.本结果对其他品种山药或富含多糖多酚类物质植物的RNA的提取也具有一定借鉴意义.展开更多
Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),the pathogen responsible for coronavirus disease 2019(COVID-19),continues to evolve,giving rise to more variants and global reinfections.Previous research ha...Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),the pathogen responsible for coronavirus disease 2019(COVID-19),continues to evolve,giving rise to more variants and global reinfections.Previous research has demonstrated that barcode segments can effectively and cost-efficiently identify specific species within closely related populations.In this study,we designed and tested RNA barcode segments based on genetic evolutionary relationships to facilitate the efficient and accurate identification of SARS-CoV-2 from extensive virus samples,including human coronaviruses(HCoVs)and SARSr-CoV-2 lineages.Nucleotide sequences sourced from NCBI and GISAID were meticulously selected and curated to construct training sets,encompassing 1733 complete genome sequences of HCoVs and SARSr-CoV-2 lineages.Through genetic-level species testing,we validated the accuracy and reliability of the barcode segments for identifying SARS-CoV-2.Subsequently,75 main and subordinate species-specific barcode segments for SARS-CoV-2,located in ORF1ab,S,E,ORF7a,and N coding sequences,were intercepted and screened based on single-nucleotide polymorphism sites and weighted scores.Post-testing,these segments exhibited high recall rates(nearly 100%),specificity(almost 30%at the nucleotide level),and precision(100%)performance on identification.They were eventually visualized using one and two-dimensional combined barcodes and deposited in an online database(http://virusbarcodedatabase.top/).The successful integration of barcoding technology in SARS-CoV-2 identification provides valuable insights for future studies involving complete genome sequence polymorphism analysis.Moreover,this cost-effective and efficient identification approach also provides valuable reference for future research endeavors related to virus surveillance.展开更多
Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding...Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus(PPRV)pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals,establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV.展开更多
Based on the Perelmann mappings from the homogenous 5D onto the inhomogeneous Lorentz 4D manifold at liquid water phase temperature range, it is shown that coherent and decoherent molecules, water, carbon, hydrogen an...Based on the Perelmann mappings from the homogenous 5D onto the inhomogeneous Lorentz 4D manifold at liquid water phase temperature range, it is shown that coherent and decoherent molecules, water, carbon, hydrogen and oxygen naturally replace the Quarks in the standard model at Bethe fusion temperature as the SU(3) generators, leading to the formation of nitrogenous bases to house the monopole boson eigenstates that can exist within the RNA and DNA. Through which the growth of organic cells by inducing the Off Diagonal Long-Range Order of “p” type hole states in organic rings and bio-cells and thus “Life”.展开更多
文摘为提取高质量的山药总RNA,以铁棍山药茎段为材料,采用Trizol法、RNAiso for Polysacchariderich Plant Tissue法、改良CTAB法和试剂盒法提取总RNA,利用琼脂糖凝胶电泳分析所提取RNA的完整性,并用超微量分光光度计分析RNA的纯度.结果显示采用Trizol法难以提RNA,RNAiso for Polysacchariderich Plant Tissue法提取效果不理想,存在DNA和蛋白质污染,而用改良CTAB法和试剂盒法提取到的总RNA纯度高、完整性好,其中,改良CTAB法较试剂盒法价格便宜,但提取过程较烦琐.因此,应根据实验需要选择CTAB法或试剂盒法提取铁棍山药总RNA.本结果对其他品种山药或富含多糖多酚类物质植物的RNA的提取也具有一定借鉴意义.
基金supported by grants from Key Research&Development Project of Nanhua Biomedical Co.,Ltd.(No.H202191490139)National Natural Science Foundation of China(No.31872866)+1 种基金China Postdoctoral Science Foundation(Nos.2021M701160 and 2022M721101)Funds of Hunan university(521119400156).
文摘Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),the pathogen responsible for coronavirus disease 2019(COVID-19),continues to evolve,giving rise to more variants and global reinfections.Previous research has demonstrated that barcode segments can effectively and cost-efficiently identify specific species within closely related populations.In this study,we designed and tested RNA barcode segments based on genetic evolutionary relationships to facilitate the efficient and accurate identification of SARS-CoV-2 from extensive virus samples,including human coronaviruses(HCoVs)and SARSr-CoV-2 lineages.Nucleotide sequences sourced from NCBI and GISAID were meticulously selected and curated to construct training sets,encompassing 1733 complete genome sequences of HCoVs and SARSr-CoV-2 lineages.Through genetic-level species testing,we validated the accuracy and reliability of the barcode segments for identifying SARS-CoV-2.Subsequently,75 main and subordinate species-specific barcode segments for SARS-CoV-2,located in ORF1ab,S,E,ORF7a,and N coding sequences,were intercepted and screened based on single-nucleotide polymorphism sites and weighted scores.Post-testing,these segments exhibited high recall rates(nearly 100%),specificity(almost 30%at the nucleotide level),and precision(100%)performance on identification.They were eventually visualized using one and two-dimensional combined barcodes and deposited in an online database(http://virusbarcodedatabase.top/).The successful integration of barcoding technology in SARS-CoV-2 identification provides valuable insights for future studies involving complete genome sequence polymorphism analysis.Moreover,this cost-effective and efficient identification approach also provides valuable reference for future research endeavors related to virus surveillance.
基金supported by the National Key Research and Development Program of China (2016YFD0500108 and 2016YFE0204100)the International Cooperation Project of CAAS Innovation Program (CAAS-GJHZ201700X)
文摘Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus(PPRV)pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals,establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV.
文摘Based on the Perelmann mappings from the homogenous 5D onto the inhomogeneous Lorentz 4D manifold at liquid water phase temperature range, it is shown that coherent and decoherent molecules, water, carbon, hydrogen and oxygen naturally replace the Quarks in the standard model at Bethe fusion temperature as the SU(3) generators, leading to the formation of nitrogenous bases to house the monopole boson eigenstates that can exist within the RNA and DNA. Through which the growth of organic cells by inducing the Off Diagonal Long-Range Order of “p” type hole states in organic rings and bio-cells and thus “Life”.
