Phytophthora root and stem rot of soybean caused by Phytophthora sojae(P.sojae)is a devastating disease that affects soybean[Glycine max(L.)Merr.]all over the world.S-phase kinase-associated protein 1(SKP1)proteins ar...Phytophthora root and stem rot of soybean caused by Phytophthora sojae(P.sojae)is a devastating disease that affects soybean[Glycine max(L.)Merr.]all over the world.S-phase kinase-associated protein 1(SKP1)proteins are key members of the SKP1/Cullin/F-box protein(SCF)ubiquitin ligase complex and play diverse roles in plant biology.However,the role of SKP1 in soybean against the phytopathogenic oomycete P.sojae remains unclear.In this study,a novel member of the soybean SKP1 gene family,GmSKP1 which was significantly induced by P.sojae,was reported.The expression of GmSKP1 was simultaneously induced by methyl jasmonate(MeJA),salicylic acid(SA)and ethylene(ET),which might suggest an important role for GmSKP1 of plant in responses to hormone treatments.Functional analysis using GmSKP1 overexpression lines showed that GmSKP1 enhanced resistance to P.sojae in transgenic soybean plants.Further analyses showed that GmSKP1 interacted with a homeodomain-leucine zipper protein transcription factor(GmHDL56)and a WRKY transcription factor(GmWRKY31),which could positively regulate responses to P.sojae in soybean.Importantly,several pathogenesis-related(PR)genes were constitutively activated,including GmPR1a,GmPR2,GmPR3,GmPR4,GmPR5a and GmPR10,in GmSKP1-OE soybean plants.Taken together,these results suggested that GmSKP1 enhanced resistance to P.sojae in soybean,possibly by activating the defense-related PR genes.展开更多
Objective:To study the inhibitory effect of siRNA mediated by expression vectors on Skp2 expression in human chondrocytes.Method:Three Skp2 sequences siRNA-59,siRNA-318 and siRNA-504 were designed as target sites usin...Objective:To study the inhibitory effect of siRNA mediated by expression vectors on Skp2 expression in human chondrocytes.Method:Three Skp2 sequences siRNA-59,siRNA-318 and siRNA-504 were designed as target sites using online siRNA design tools.Skp2 siRNA expression vectors were successfully constructed in vitro by gene recombination technology,and the influence of recombinant plasmid transfection on Skp2 mRNA expression was detected.DNA electrophoresis was used to verify the results.Results:The sequence of Skp2 interference was correct by sequence analysis.The expression of Skp2 mRNA in siRNA-59,siRNA-318,siRNA-504 transfection group was significantly lower than that in no-load group and NC group(P<0.05),the inhibition rates of Skp2 mRNA in siRNA-59,siRNA-318 and siRNA-504 were respectively 60%,41%and 64%,and the siRNA-504 transfection group had the highest inhibition rate.Conclusion:The siRNA eukaryotic expression vector of Skp2 gene was constructed successfully which effectively inhibit Skp2 mRNA expression in human chondrocytes cell,and can provide strong evidence for the treatment of osteoarthritis.展开更多
背景与目的S期激酶相关蛋白2(Skp2)是细胞周期正性调节因子之一,它能促进周期蛋白依赖性激酶抑制剂p27的泛素化蛋白水解,在肿瘤中过表达,本研究旨在探讨非小细胞肺癌(non-small cell lung cancer NSCLC)中Skp2表达的临床意义及其与p27...背景与目的S期激酶相关蛋白2(Skp2)是细胞周期正性调节因子之一,它能促进周期蛋白依赖性激酶抑制剂p27的泛素化蛋白水解,在肿瘤中过表达,本研究旨在探讨非小细胞肺癌(non-small cell lung cancer NSCLC)中Skp2表达的临床意义及其与p27蛋白表达的关系。方法应用组织芯片和免疫组织化学方法检测Skp2和p27在68例NSCLC组织和17例正常支气管上皮细胞中的表达。结果Skp2仅在肺癌组织中表达,且与患者的组织学类型(P=0.039),肿瘤细胞的分化程度(P=0.016),性别(P=0.012)和吸烟与否(P=0.026)显著相关,而与患者的年龄和TNM分期无关。p27在正常支气管上皮细胞中均有表达,在肺癌组织中表达降低;Skp2阳性表达的患者中p27表达明显降低,两者呈负相关(P=0.021)。结论在NSCLC中,Skp2蛋白表达的增高与p27泛素化依赖的蛋白降解有关,提示Skp2蛋白过表达在NSCLC的发生和发展中可能起重要作用。展开更多
基金Supported by the NSFC Projects(31971972)the Natural Science Foundation of Heilongjiang Province(ZD2019C001)the Outstanding Talents and Innovative Team of Agricultural Scientific Research。
文摘Phytophthora root and stem rot of soybean caused by Phytophthora sojae(P.sojae)is a devastating disease that affects soybean[Glycine max(L.)Merr.]all over the world.S-phase kinase-associated protein 1(SKP1)proteins are key members of the SKP1/Cullin/F-box protein(SCF)ubiquitin ligase complex and play diverse roles in plant biology.However,the role of SKP1 in soybean against the phytopathogenic oomycete P.sojae remains unclear.In this study,a novel member of the soybean SKP1 gene family,GmSKP1 which was significantly induced by P.sojae,was reported.The expression of GmSKP1 was simultaneously induced by methyl jasmonate(MeJA),salicylic acid(SA)and ethylene(ET),which might suggest an important role for GmSKP1 of plant in responses to hormone treatments.Functional analysis using GmSKP1 overexpression lines showed that GmSKP1 enhanced resistance to P.sojae in transgenic soybean plants.Further analyses showed that GmSKP1 interacted with a homeodomain-leucine zipper protein transcription factor(GmHDL56)and a WRKY transcription factor(GmWRKY31),which could positively regulate responses to P.sojae in soybean.Importantly,several pathogenesis-related(PR)genes were constitutively activated,including GmPR1a,GmPR2,GmPR3,GmPR4,GmPR5a and GmPR10,in GmSKP1-OE soybean plants.Taken together,these results suggested that GmSKP1 enhanced resistance to P.sojae in soybean,possibly by activating the defense-related PR genes.
基金National Natural Science Foundation of China(82060875)National Natural Science Foundation of China(82160912)Guangxi Natural Science Foundation(2022JJA141229)。
文摘Objective:To study the inhibitory effect of siRNA mediated by expression vectors on Skp2 expression in human chondrocytes.Method:Three Skp2 sequences siRNA-59,siRNA-318 and siRNA-504 were designed as target sites using online siRNA design tools.Skp2 siRNA expression vectors were successfully constructed in vitro by gene recombination technology,and the influence of recombinant plasmid transfection on Skp2 mRNA expression was detected.DNA electrophoresis was used to verify the results.Results:The sequence of Skp2 interference was correct by sequence analysis.The expression of Skp2 mRNA in siRNA-59,siRNA-318,siRNA-504 transfection group was significantly lower than that in no-load group and NC group(P<0.05),the inhibition rates of Skp2 mRNA in siRNA-59,siRNA-318 and siRNA-504 were respectively 60%,41%and 64%,and the siRNA-504 transfection group had the highest inhibition rate.Conclusion:The siRNA eukaryotic expression vector of Skp2 gene was constructed successfully which effectively inhibit Skp2 mRNA expression in human chondrocytes cell,and can provide strong evidence for the treatment of osteoarthritis.
文摘背景与目的S期激酶相关蛋白2(Skp2)是细胞周期正性调节因子之一,它能促进周期蛋白依赖性激酶抑制剂p27的泛素化蛋白水解,在肿瘤中过表达,本研究旨在探讨非小细胞肺癌(non-small cell lung cancer NSCLC)中Skp2表达的临床意义及其与p27蛋白表达的关系。方法应用组织芯片和免疫组织化学方法检测Skp2和p27在68例NSCLC组织和17例正常支气管上皮细胞中的表达。结果Skp2仅在肺癌组织中表达,且与患者的组织学类型(P=0.039),肿瘤细胞的分化程度(P=0.016),性别(P=0.012)和吸烟与否(P=0.026)显著相关,而与患者的年龄和TNM分期无关。p27在正常支气管上皮细胞中均有表达,在肺癌组织中表达降低;Skp2阳性表达的患者中p27表达明显降低,两者呈负相关(P=0.021)。结论在NSCLC中,Skp2蛋白表达的增高与p27泛素化依赖的蛋白降解有关,提示Skp2蛋白过表达在NSCLC的发生和发展中可能起重要作用。
