Newcastle disease( ND) is one of the most serious infectious diseases that infect the poultry industry.There is only one serotype of Newcastle disease virus( NDV),but NDVs can be divided into two distinct classes( cla...Newcastle disease( ND) is one of the most serious infectious diseases that infect the poultry industry.There is only one serotype of Newcastle disease virus( NDV),but NDVs can be divided into two distinct classes( class Ⅰ,and class Ⅱ) according to their genetic relationship.To develop a method for rapid quantitative detection of class Ⅰ NDV,a pair of primers and a TaqM an probe were designed and synthesized according to the conservative sequence of NP gene of class Ⅰ NDV.The positive recombinant plasmid harboring NP gene of JS-18-05 isolate was used as a positive template to establish the standard curve.A real-time fluorescent quantitative RT-PCR method was established for rapid detection of class Ⅰ NDV with strong specificity,high sensitivity and good repeatability.The established method exhibited a good linear relationship within the concentration of 102 to 108 copies of NDV,by which 1 μl of 10 copy of NDV nucleic acid could be detected in the initial template.Compared with conventional virus isolation methods,the established method had similar sensitivity and led to the same results in detecting33 class Ⅰ,class Ⅱ NDV isolates.The study provided the basis for rapid quantitative detection of class Ⅰ NDVs and further clarification of their pathogenicity and pathogenic mechanism in poultry.展开更多
A digital RT-PCR method for rapid detection of H9 subtype influenza was established by comparing the two methods of digital RT-PCR and real-time quantitative RT-PCR. The sensitivity, specificity and reproducibility of...A digital RT-PCR method for rapid detection of H9 subtype influenza was established by comparing the two methods of digital RT-PCR and real-time quantitative RT-PCR. The sensitivity, specificity and reproducibility of the two methods for H9 were determined by gradient dilution using the same pair of primers and probes. Both methods were able to detect 104 times diluted H9 pathogens, while digital RT-PCR could detect H9 in single droplets, and its sensitivity was higher than real-time quantitative RT-PCR. At the same time, the specificities of both methods were very strong, with no amplification reactions for H3N2, H4N2, H6N2. The reproducibility of the two methods were also good. Digital RT-PCR has a higher sensitivity than real-time quantitative RT-PCR and could play an important role in the rapid detection of H9 subtype influenza virus.展开更多
Objective:To establish a new detecting method for disease susceptibility loci R1628P and G2385R of Parkinson’s disease(PD)related gene LRRK2.Methods:Sequence specific primers were designed to make a genotyping of DNA...Objective:To establish a new detecting method for disease susceptibility loci R1628P and G2385R of Parkinson’s disease(PD)related gene LRRK2.Methods:Sequence specific primers were designed to make a genotyping of DNA markers with known genotypes by use of quantitative fluorescence real-time PCR(RT-PCR).100 cases of PD samples with unknown genotypes were tested,and verified by use of polymerase chain reaction linked restriction fragment length polymorphism(PCR-RLFP).Results:The genotyping results of DNA markers proved to be correct,and 100 cases of samples to be tested had a completely consistent genotyping result with PCR-RLFP genotyping result.Conclusions:Sequence specific primer and quantitative fluorescence RT-PCR can successfully make a genotyping for disease susceptibility loci R1628P and G2385R of LRRK2.展开更多
Heat shock proteins (HSPs), as molecular chaperones, play an important role under physiological condition and in the course of many diseases. It would therefore be valuable to determine the expression of cellular hsp ...Heat shock proteins (HSPs), as molecular chaperones, play an important role under physiological condition and in the course of many diseases. It would therefore be valuable to determine the expression of cellular hsp gene quantitatively. Using DNA recombinant technique and in vitro transcription system, a complex internal control RNA has been prepared. After opti-展开更多
Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 1...Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).展开更多
Cells in mammalian cochleae virtually stop proliferation and exit cellular circle before birth. Consequently, hair cells and spiral ganglion neurons destroyed by ototoxic factors cannot be replaced through proliferati...Cells in mammalian cochleae virtually stop proliferation and exit cellular circle before birth. Consequently, hair cells and spiral ganglion neurons destroyed by ototoxic factors cannot be replaced through proliferative regeneration. However, substantial proliferation occurs in organotypic cultures of cochleae from postnatal mice. In the present study, we studied the time course of proliferative growth in cultures of mouse cochlea explants obtained from up to 12 postnatal days. The mitotic nature of this growth was confirmed by bromodeoxyuridine (BrdU) staining and expression of proliferation cell nuclear antigen (PCNA) evaluated with real-time quantitative poly-merase chain reaction(RT-PCR). Similar growth time course was found in the cochlear explants of different postnatal ages. The new growth reached its maximum at around 2 days in culture followed by a slow-down, and virtually stopped after 5 days of culture. The possible mechanisms and the significance of this proliferation are discussed.展开更多
Onion(Allium cepa L.) was an economic vegetable and a strictly biennial herb, which was widely distributed in the world. In the past, it was a strictly biennial plant, the studies had shown that FT(FLOWERING LOCUS T) ...Onion(Allium cepa L.) was an economic vegetable and a strictly biennial herb, which was widely distributed in the world. In the past, it was a strictly biennial plant, the studies had shown that FT(FLOWERING LOCUS T) gene was involved in the photoperiod pathway to regulate flowering in the model plant. In this study, transcriptome sequencing method was used to obtain cDNA sequence of FT homologous gene in onion, named AcFT3(KF864665). AcFT3 had a full-length of 540 bp, encoded 179 amino acids, with 98.31% homology to AfFT(Allium fistulosum), and 63.0%-84.0% homology to other higher plants. Phylogenetic tree analysis indicated that AcFT3 had the closest relationship with AfFT. The results of quantitative RT-PCR showed the expression pattern of AcFT3 both in vegetative growth of onion and in different organs of bolting and flowering, the expression level of AcFT3 reached the highest in the leaves before bolting and in the flower organs after bolting.展开更多
Cytoplasmic male sterility(CMS)-restorer system is a useful tool to exploit heterosis in soybean.The major restorer gene for the M-type CMS is known as Rf-m,located in the 162.4-kb region on chromosome 16.Sequence ana...Cytoplasmic male sterility(CMS)-restorer system is a useful tool to exploit heterosis in soybean.The major restorer gene for the M-type CMS is known as Rf-m,located in the 162.4-kb region on chromosome 16.Sequence analysis has revealed that the Rf-m locus in Glycine max consists of seven penta tricopeptide repeat(GmPPR)genes.The deduced amino acid sequences contain 8 to 14 PPR motifs,and a phylogenetic analysis grouped these GmPPR proteins into two PPR subfamilies:Glyma.16G161800 belongs to the PLS subfamily,and the P subfamily consists.of Glyma.16G161900,Glyma 16G162000,Glyma.16G162100,Glyma.16G162700,Glyma.16G162800,and Gly-ma 16G163100.The phylogenetic analysis of seven GmPPR proteins and 27 other plant PPR proteins also showed that proteins in the same subfamilies cluster together.Comparative sequence analysis was conducted using the seven Rf-m candidate GmPPR genes from the sterile line W931A,the maintainer line W931B,and the restorer line WR016,the result showed that Glyma 16G161900 had higher polymorphism than the other candidate genes.Based on real-time quantitative RT-PCR data,all seven GmPPR genes were differentially expressed but showed constitutive expression in roots,stems,leaves,and pollen grains.Additionally,the expression level of Gly-ma 16G161900 in the sterile line W931 A was significantly higher in all tissues than in the restorer line WR016.Taken together,these results suggest that Glyma 16G161900 is the most likely candidate for the restorer gene Rf-m.This study is the first report and analysis of candidate fertility restorer(Rf)genes encoding PPR proteins in soybean.展开更多
Plant microRNAs(miRNAs)play important roles in biological processes such as development and stress responses.Although the diverse functions of miRNAs in model organisms have been well studied,their function in wild ri...Plant microRNAs(miRNAs)play important roles in biological processes such as development and stress responses.Although the diverse functions of miRNAs in model organisms have been well studied,their function in wild rice is poorly understood.In this study,high-throughput small RNA sequencing was performed to characterize tissue-specific transcriptomes in Oryza longistaminata.A total of 603 miRNAs,380 known rice miRNAs,72 conserved plant miRNAs,and151 predicted novel miRNAs were identified as being expressed in aerial shoots and rhizomes.Additionally,99 and 79 miRNAs were expressed exclusively or differentially,respectively,in the two tissues,and 144 potential targets were predicted for the differentially expressed miRNAs in the rhizomes.Functional annotation of these targets suggested that transcription factors,including squamosa promoter binding proteins and auxin response factors,function in rhizome growth and development.The expression levels of several miRNAs and target genes in the rhizomes were quantified by RT-PCR,and the results indicated the existence of complex regulatory mechanisms between the miRNAs and their targets.Eight target cleavage sites were verified by RNA ligase-mediated rapid 5′end amplification.These results provide valuable information on the composition,expression and function of miRNAs in O.longistaminata,and will aid in understanding the molecular mechanisms of rhizome development.展开更多
中国蜂蜜主要分为中蜂蜂蜜和意蜂蜂蜜两种,分别由中蜂和意蜂酿造而成。中蜂蜂蜜营养价值高,保健功能好,但是由于产量低,价格是意蜂蜂蜜的3~5倍。一些不法企业和养蜂人趁机将意蜂蜂蜜假冒中蜂蜂蜜销售,或将意蜂蜂蜜掺入到中蜂蜂蜜中以次...中国蜂蜜主要分为中蜂蜂蜜和意蜂蜂蜜两种,分别由中蜂和意蜂酿造而成。中蜂蜂蜜营养价值高,保健功能好,但是由于产量低,价格是意蜂蜂蜜的3~5倍。一些不法企业和养蜂人趁机将意蜂蜂蜜假冒中蜂蜂蜜销售,或将意蜂蜂蜜掺入到中蜂蜂蜜中以次充好。因此,如何鉴别中蜂蜂蜜和意蜂蜂蜜是当前蜂产品行业面临的一大难题。本研究以MRJP2(Major Royal Jelly Protein 2)为靶标基因,分别设计了针对中蜂和意蜂的特异性引物和探针,开发了实时荧光PCR-Taqman探针法,并对该方法的特异性和灵敏度进行了分析。结果显示该方法可以特异性的区分中蜂蜂蜜和意蜂蜂蜜;对意蜂蜂蜜DNA的最低检测限达到0.05ng,对中蜂蜂蜜DNA的最低检测限达到0.01ng;对市售中蜂蜂蜜进行真实性摸底调查发现约37%样品为中蜂蜂蜜,其余样品均含有意蜂蜂蜜成分。综上所述,中蜂蜂蜜掺假现象严重,实时荧光PCR方法检测灵敏度高、特异性强、操作简便快速,可为中蜂蜂蜜掺假问题提供技术支持。展开更多
Objective: To investigate the antiviral property of a lead ligand, YK51 that was synthesized based on the flavanoid of a natural product toward dengue virus type-2(DENV2)replication.Methods: c RNA was isolated from He...Objective: To investigate the antiviral property of a lead ligand, YK51 that was synthesized based on the flavanoid of a natural product toward dengue virus type-2(DENV2)replication.Methods: c RNA was isolated from HepG2 cells inoculated with 1 000 median tissue culture infective dose of DENV2 and treated with different doses of the ligand followed by RT-PCR to quantify the virus gene copies. Confocal microscopy of actin and tubulin redistribution was also performed.Results: The quantitative RT-PCR result showed reduction of the DENV2 gene copies as the ligand concentration was increased. The confocal microscopy result showed increase in the tubulin intensity(79.6%) of infected BHK21 cells treated with the ligand,compared with the non-treated cells(54.8%). The 1.5-fold increase in the intensity of tubulin suggested that the ligand inhibitory effect stabilized the cellular microtubule structure.Conclusions: The synthesized ligand YK51 reduced DENV2 viral load by inhibiting virus replication thus is highly potential to be developed as antiviral agent.展开更多
<strong>Objective: </strong><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">To investigate the prote...<strong>Objective: </strong><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">To investigate the protein expression and clinicopathological characteristics of ABCG2</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">, </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">Oct4</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">,</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> Nanog in laryngeal cancer tissues, and to seek new molecular markers for the diagnosis of laryngeal cancer.</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><b><span style="font-family:Verdana;">Methods: </span></b></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">The laryngeal cancer tissues and paracancerous tissues of 87 patients with laryngeal carcinoma diagnosed in the department of otorhinolaryngology and head and neck surgery in our hospital from April 2016 to April 2018 were selected as the subjects. QRT-PCR, Real-time PcR, Western blot and immunohistochemical staining were used to detect the expression of ABCG2, Oct4 and Nanog in (Tumor Tissue) and (Adjacent Tissue) in tumor tissue and paracancerous tissue.</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><b><span style="font-family:Verdana;">Results: </span></b></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">The results of RT-PCR showed that the positive rates of ABCG2, Oct4 and Nanog in laryngeal carcinoma tissues were 49.30%, 45.07% and 52.11%, respectively, while those in paracancerous tissues were </span><span style="font-family:Verdana;">22.54%, 21.13% and 15.49%, respectively (P < 0.01). The expression of ABCG2,</span><span style="font-family:Verdana;"> Oct4 and Nanog in laryngeal carcinoma was correlated with tumor differentiation, depth of invasion, age and sex (P < 0.05), but not with tumor size and TNM stage.</span></span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><b><span style="font-family:Verdana;">Conclusion: </span></b></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">The expressions of ABCG2, Oct4 and Nanog in cancer tissues are related to tumor differentiation status, and they can be used as new molecular markers for the diagnosis of laryngeal cancer</span></span></span><span style="font-family:Verdana;">.</span>展开更多
It has been hypothesized that Rab3A, a small GTPase, may be closely involved in the process of dense core vesicle exocytosis in various cell types. This possibility was investigated by disrupting the expression levels...It has been hypothesized that Rab3A, a small GTPase, may be closely involved in the process of dense core vesicle exocytosis in various cell types. This possibility was investigated by disrupting the expression levels of Rab3A-mRNA using a small interfering RNA of the Rab3A GTPase (Rab3A-siRNA) and examining the effect of this on transcytosis of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP). Rab3A-siRNA and WGA-HRP were injected into the right vagus nerves of adult rats which were killed 12, 24 or 48 hours later. In some animals, portions of the brain stem containing the nucleus of solitary tract (NST) were prepared for electron microscopy. In other animals, the nodose ganglion of the vagus nerve was used to determine the levels of expression of Rab3A-mRNA using RT-PCR techniques. It was found that the expression of Rab3A-mRNA was markedly depressed in animals at 12 h after the Rab3A-siRNA injection. In the NST, there was an accumulation of HRP-reaction product (RP), recognized as electron dense lysosomal-like structures, in both axons and terminals in the NST 12 h after injection. Some HRP-RP was found in membrane bound vesicles in close proximity to cell membranes and appeared to be in the process of transcytosis. This neuronal transcytosis of HRP-RP appeared to occur at random locations over the axodendritic membranes. These findings indicate that inhibiting the expression of Rab3A-mRNA using Rab3A-siRNA can modulate the level of transcytosis of proteins across neuronal membranes confirming the potentially important role of this GTPase in the process of transcytosis.展开更多
基金Supported by National Natural Science Foundation of China(30630048)National Science and Technology Support Program(2006BAD06A03)
文摘Newcastle disease( ND) is one of the most serious infectious diseases that infect the poultry industry.There is only one serotype of Newcastle disease virus( NDV),but NDVs can be divided into two distinct classes( class Ⅰ,and class Ⅱ) according to their genetic relationship.To develop a method for rapid quantitative detection of class Ⅰ NDV,a pair of primers and a TaqM an probe were designed and synthesized according to the conservative sequence of NP gene of class Ⅰ NDV.The positive recombinant plasmid harboring NP gene of JS-18-05 isolate was used as a positive template to establish the standard curve.A real-time fluorescent quantitative RT-PCR method was established for rapid detection of class Ⅰ NDV with strong specificity,high sensitivity and good repeatability.The established method exhibited a good linear relationship within the concentration of 102 to 108 copies of NDV,by which 1 μl of 10 copy of NDV nucleic acid could be detected in the initial template.Compared with conventional virus isolation methods,the established method had similar sensitivity and led to the same results in detecting33 class Ⅰ,class Ⅱ NDV isolates.The study provided the basis for rapid quantitative detection of class Ⅰ NDVs and further clarification of their pathogenicity and pathogenic mechanism in poultry.
文摘A digital RT-PCR method for rapid detection of H9 subtype influenza was established by comparing the two methods of digital RT-PCR and real-time quantitative RT-PCR. The sensitivity, specificity and reproducibility of the two methods for H9 were determined by gradient dilution using the same pair of primers and probes. Both methods were able to detect 104 times diluted H9 pathogens, while digital RT-PCR could detect H9 in single droplets, and its sensitivity was higher than real-time quantitative RT-PCR. At the same time, the specificities of both methods were very strong, with no amplification reactions for H3N2, H4N2, H6N2. The reproducibility of the two methods were also good. Digital RT-PCR has a higher sensitivity than real-time quantitative RT-PCR and could play an important role in the rapid detection of H9 subtype influenza virus.
文摘Objective:To establish a new detecting method for disease susceptibility loci R1628P and G2385R of Parkinson’s disease(PD)related gene LRRK2.Methods:Sequence specific primers were designed to make a genotyping of DNA markers with known genotypes by use of quantitative fluorescence real-time PCR(RT-PCR).100 cases of PD samples with unknown genotypes were tested,and verified by use of polymerase chain reaction linked restriction fragment length polymorphism(PCR-RLFP).Results:The genotyping results of DNA markers proved to be correct,and 100 cases of samples to be tested had a completely consistent genotyping result with PCR-RLFP genotyping result.Conclusions:Sequence specific primer and quantitative fluorescence RT-PCR can successfully make a genotyping for disease susceptibility loci R1628P and G2385R of LRRK2.
文摘Heat shock proteins (HSPs), as molecular chaperones, play an important role under physiological condition and in the course of many diseases. It would therefore be valuable to determine the expression of cellular hsp gene quantitatively. Using DNA recombinant technique and in vitro transcription system, a complex internal control RNA has been prepared. After opti-
基金partially supported by the National Institutes of Health(grant no.P20GM103646)the United States Department of Agriculture Animal and Plant Health Inspection Service(agreement 14-7428-1041-CA)
文摘Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).
文摘该研究旨在建立一种快速、敏感和特异性检测新型鸭呼肠孤病毒（Novel duck reovirus,NDRV）的荧光定量PCR诊断方法。本实验以感染新型鸭呼肠孤病毒的鸭组织RNA提取物为模板,根据Gen Bank数据库中呼肠孤病毒S1基因全序列,设计合成了一对特异性引物,PCR扩增基因片段,将其克隆至p ET-30a载体,重组质粒测序并进行同源性分析;以阳性质粒为模板,建立SYBR Green II荧光定量PCR检测方法,并进行敏感性和特异性检测。经测序证实扩增片段与预期目的片段相符,所建立的SYBR Green II荧光定量PCR检测S1的反应在101～108 copies/u L之间具有良好的线性关系,反应的检出下限为10 copies/μL,而H5型禽流感病毒、H9型禽流感病毒、鸡传染性支气管炎病毒、C型鸭肝炎病毒、新城疫病毒、鹅细小病毒、鸭瘟病毒等病毒的检测为阴性,表明该方法敏感、特异。本研究成功建立了SYBR Green II荧光定量PCR检测新型鸭呼肠孤病毒的方法,为新型鸭呼肠孤病毒致病机制和机体免疫保护机制的研究提供了技术平台。
文摘Cells in mammalian cochleae virtually stop proliferation and exit cellular circle before birth. Consequently, hair cells and spiral ganglion neurons destroyed by ototoxic factors cannot be replaced through proliferative regeneration. However, substantial proliferation occurs in organotypic cultures of cochleae from postnatal mice. In the present study, we studied the time course of proliferative growth in cultures of mouse cochlea explants obtained from up to 12 postnatal days. The mitotic nature of this growth was confirmed by bromodeoxyuridine (BrdU) staining and expression of proliferation cell nuclear antigen (PCNA) evaluated with real-time quantitative poly-merase chain reaction(RT-PCR). Similar growth time course was found in the cochlear explants of different postnatal ages. The new growth reached its maximum at around 2 days in culture followed by a slow-down, and virtually stopped after 5 days of culture. The possible mechanisms and the significance of this proliferation are discussed.
基金Supported by Fund Project of Heilongjiang Province(C2015017)Heilongjiang Provincial Science and Technology Commissioner Project(GC13B09)
文摘Onion(Allium cepa L.) was an economic vegetable and a strictly biennial herb, which was widely distributed in the world. In the past, it was a strictly biennial plant, the studies had shown that FT(FLOWERING LOCUS T) gene was involved in the photoperiod pathway to regulate flowering in the model plant. In this study, transcriptome sequencing method was used to obtain cDNA sequence of FT homologous gene in onion, named AcFT3(KF864665). AcFT3 had a full-length of 540 bp, encoded 179 amino acids, with 98.31% homology to AfFT(Allium fistulosum), and 63.0%-84.0% homology to other higher plants. Phylogenetic tree analysis indicated that AcFT3 had the closest relationship with AfFT. The results of quantitative RT-PCR showed the expression pattern of AcFT3 both in vegetative growth of onion and in different organs of bolting and flowering, the expression level of AcFT3 reached the highest in the leaves before bolting and in the flower organs after bolting.
基金the National Key Research and Development Program of China(Grant No.2016YFD0101503)the Key Research and Development Program of Anhui Province(Grant No.202004a06020034)+1 种基金the Major Science and Technology Project of Anhui Province(Grant No.18030701178)the Program on Industrial Technology System of National Soybean(Grant No.CARS-04-PS07)。
文摘Cytoplasmic male sterility(CMS)-restorer system is a useful tool to exploit heterosis in soybean.The major restorer gene for the M-type CMS is known as Rf-m,located in the 162.4-kb region on chromosome 16.Sequence analysis has revealed that the Rf-m locus in Glycine max consists of seven penta tricopeptide repeat(GmPPR)genes.The deduced amino acid sequences contain 8 to 14 PPR motifs,and a phylogenetic analysis grouped these GmPPR proteins into two PPR subfamilies:Glyma.16G161800 belongs to the PLS subfamily,and the P subfamily consists.of Glyma.16G161900,Glyma 16G162000,Glyma.16G162100,Glyma.16G162700,Glyma.16G162800,and Gly-ma 16G163100.The phylogenetic analysis of seven GmPPR proteins and 27 other plant PPR proteins also showed that proteins in the same subfamilies cluster together.Comparative sequence analysis was conducted using the seven Rf-m candidate GmPPR genes from the sterile line W931A,the maintainer line W931B,and the restorer line WR016,the result showed that Glyma 16G161900 had higher polymorphism than the other candidate genes.Based on real-time quantitative RT-PCR data,all seven GmPPR genes were differentially expressed but showed constitutive expression in roots,stems,leaves,and pollen grains.Additionally,the expression level of Gly-ma 16G161900 in the sterile line W931 A was significantly higher in all tissues than in the restorer line WR016.Taken together,these results suggest that Glyma 16G161900 is the most likely candidate for the restorer gene Rf-m.This study is the first report and analysis of candidate fertility restorer(Rf)genes encoding PPR proteins in soybean.
基金supported by the National Natural Science Foundation of China(31271694 and U1302264)
文摘Plant microRNAs(miRNAs)play important roles in biological processes such as development and stress responses.Although the diverse functions of miRNAs in model organisms have been well studied,their function in wild rice is poorly understood.In this study,high-throughput small RNA sequencing was performed to characterize tissue-specific transcriptomes in Oryza longistaminata.A total of 603 miRNAs,380 known rice miRNAs,72 conserved plant miRNAs,and151 predicted novel miRNAs were identified as being expressed in aerial shoots and rhizomes.Additionally,99 and 79 miRNAs were expressed exclusively or differentially,respectively,in the two tissues,and 144 potential targets were predicted for the differentially expressed miRNAs in the rhizomes.Functional annotation of these targets suggested that transcription factors,including squamosa promoter binding proteins and auxin response factors,function in rhizome growth and development.The expression levels of several miRNAs and target genes in the rhizomes were quantified by RT-PCR,and the results indicated the existence of complex regulatory mechanisms between the miRNAs and their targets.Eight target cleavage sites were verified by RNA ligase-mediated rapid 5′end amplification.These results provide valuable information on the composition,expression and function of miRNAs in O.longistaminata,and will aid in understanding the molecular mechanisms of rhizome development.
文摘中国蜂蜜主要分为中蜂蜂蜜和意蜂蜂蜜两种,分别由中蜂和意蜂酿造而成。中蜂蜂蜜营养价值高,保健功能好,但是由于产量低,价格是意蜂蜂蜜的3~5倍。一些不法企业和养蜂人趁机将意蜂蜂蜜假冒中蜂蜂蜜销售,或将意蜂蜂蜜掺入到中蜂蜂蜜中以次充好。因此,如何鉴别中蜂蜂蜜和意蜂蜂蜜是当前蜂产品行业面临的一大难题。本研究以MRJP2(Major Royal Jelly Protein 2)为靶标基因,分别设计了针对中蜂和意蜂的特异性引物和探针,开发了实时荧光PCR-Taqman探针法,并对该方法的特异性和灵敏度进行了分析。结果显示该方法可以特异性的区分中蜂蜂蜜和意蜂蜂蜜;对意蜂蜂蜜DNA的最低检测限达到0.05ng,对中蜂蜂蜜DNA的最低检测限达到0.01ng;对市售中蜂蜂蜜进行真实性摸底调查发现约37%样品为中蜂蜂蜜,其余样品均含有意蜂蜂蜜成分。综上所述,中蜂蜂蜜掺假现象严重,实时荧光PCR方法检测灵敏度高、特异性强、操作简便快速,可为中蜂蜂蜜掺假问题提供技术支持。
基金Supported by Science Fund from the Ministry of Science,Technology and Innovation Malaysia and Research Acculturation Grants of Universiti Teknologi MARA(UiTM)[RAGS/2012/Ui TM/ST04/1],Malaysia
文摘Objective: To investigate the antiviral property of a lead ligand, YK51 that was synthesized based on the flavanoid of a natural product toward dengue virus type-2(DENV2)replication.Methods: c RNA was isolated from HepG2 cells inoculated with 1 000 median tissue culture infective dose of DENV2 and treated with different doses of the ligand followed by RT-PCR to quantify the virus gene copies. Confocal microscopy of actin and tubulin redistribution was also performed.Results: The quantitative RT-PCR result showed reduction of the DENV2 gene copies as the ligand concentration was increased. The confocal microscopy result showed increase in the tubulin intensity(79.6%) of infected BHK21 cells treated with the ligand,compared with the non-treated cells(54.8%). The 1.5-fold increase in the intensity of tubulin suggested that the ligand inhibitory effect stabilized the cellular microtubule structure.Conclusions: The synthesized ligand YK51 reduced DENV2 viral load by inhibiting virus replication thus is highly potential to be developed as antiviral agent.
文摘<strong>Objective: </strong><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">To investigate the protein expression and clinicopathological characteristics of ABCG2</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">, </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">Oct4</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">,</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> Nanog in laryngeal cancer tissues, and to seek new molecular markers for the diagnosis of laryngeal cancer.</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><b><span style="font-family:Verdana;">Methods: </span></b></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">The laryngeal cancer tissues and paracancerous tissues of 87 patients with laryngeal carcinoma diagnosed in the department of otorhinolaryngology and head and neck surgery in our hospital from April 2016 to April 2018 were selected as the subjects. QRT-PCR, Real-time PcR, Western blot and immunohistochemical staining were used to detect the expression of ABCG2, Oct4 and Nanog in (Tumor Tissue) and (Adjacent Tissue) in tumor tissue and paracancerous tissue.</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><b><span style="font-family:Verdana;">Results: </span></b></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">The results of RT-PCR showed that the positive rates of ABCG2, Oct4 and Nanog in laryngeal carcinoma tissues were 49.30%, 45.07% and 52.11%, respectively, while those in paracancerous tissues were </span><span style="font-family:Verdana;">22.54%, 21.13% and 15.49%, respectively (P < 0.01). The expression of ABCG2,</span><span style="font-family:Verdana;"> Oct4 and Nanog in laryngeal carcinoma was correlated with tumor differentiation, depth of invasion, age and sex (P < 0.05), but not with tumor size and TNM stage.</span></span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><b><span style="font-family:Verdana;">Conclusion: </span></b></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">The expressions of ABCG2, Oct4 and Nanog in cancer tissues are related to tumor differentiation status, and they can be used as new molecular markers for the diagnosis of laryngeal cancer</span></span></span><span style="font-family:Verdana;">.</span>
文摘It has been hypothesized that Rab3A, a small GTPase, may be closely involved in the process of dense core vesicle exocytosis in various cell types. This possibility was investigated by disrupting the expression levels of Rab3A-mRNA using a small interfering RNA of the Rab3A GTPase (Rab3A-siRNA) and examining the effect of this on transcytosis of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP). Rab3A-siRNA and WGA-HRP were injected into the right vagus nerves of adult rats which were killed 12, 24 or 48 hours later. In some animals, portions of the brain stem containing the nucleus of solitary tract (NST) were prepared for electron microscopy. In other animals, the nodose ganglion of the vagus nerve was used to determine the levels of expression of Rab3A-mRNA using RT-PCR techniques. It was found that the expression of Rab3A-mRNA was markedly depressed in animals at 12 h after the Rab3A-siRNA injection. In the NST, there was an accumulation of HRP-reaction product (RP), recognized as electron dense lysosomal-like structures, in both axons and terminals in the NST 12 h after injection. Some HRP-RP was found in membrane bound vesicles in close proximity to cell membranes and appeared to be in the process of transcytosis. This neuronal transcytosis of HRP-RP appeared to occur at random locations over the axodendritic membranes. These findings indicate that inhibiting the expression of Rab3A-mRNA using Rab3A-siRNA can modulate the level of transcytosis of proteins across neuronal membranes confirming the potentially important role of this GTPase in the process of transcytosis.
