Calmodulin(CaM)proteins play a key role in signal transduction under various stresses.In the present study,the effects of a sugarcane ScCaM gene(NCBI accession number:GQ246454)on drought and salt stress tolerance in t...Calmodulin(CaM)proteins play a key role in signal transduction under various stresses.In the present study,the effects of a sugarcane ScCaM gene(NCBI accession number:GQ246454)on drought and salt stress tolerance in transgenic Arabidopsis thaliana and Escherichia coli cells were evaluated.The results demonstrated a significant negative role of ScCaM in the drought and salt stress tolerance of transgenic lines of A.thaliana,as indicated by the phenotypes.In addition,the expression of AtP5CS and AtRD29A,two genes tightly related to stress resistance,was significantly lower in the overexpression lines than in the wild type.The growth of E.coli BL21 cells expressing ScCaM showed weaker tolerance under mannitol and NaCl stress.Taken together,this study revealed that the ScCaM gene plays a negative regulatory role in both mannitol and NaCl stresses,and it possibly exerts protective mechanisms common in both prokaryotes and eukaryotes under stress conditions.展开更多
We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 c...We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 cells. After treatment with 1 mM db-cAMP for 1 hour and 2 hours, there was an early and repldly reduced in gene expression of Calmodulin and c-fos respectively. After db-cAMP treatment for 4 -5 days, the number of capping cells of ConA binding decreased significantly and the cell surface microvllll decreased as well. The growth of treated cells was inhibited markedly. By using 4F1 cDNA probe, which is preferentially expressed In G1 phase, we have found that the db- cAMP treated cells were accumulated at G1 phase. Of particular interest is the fact that the distribution of microtubules, mlcrofilaments and fibronectln were recovered after treatment with 1 mM db-cAMP for 6 days. It is suggested that the Inhibition of proliferation, alteration, of phenotype and reco- very of cytoskeleton is transformed cells after treatment with db-cAMP are related to the Inhibition of gene expression of Calmodulin.展开更多
Calmodulin is a calcium binding protein that modulates the activity of diverse groups of protein including some protein kinase, adenylate cyclases and ATPase. Here we use the total DNA of Sonneratia paracaseolaris as ...Calmodulin is a calcium binding protein that modulates the activity of diverse groups of protein including some protein kinase, adenylate cyclases and ATPase. Here we use the total DNA of Sonneratia paracaseolaris as the template of the polymerase chain reaction (PCR). The PCR primers have been designed and synthesized according to the 5-and 3-terminal oligonucleotide sequences of Calmodulin gene of plants in Genbank and ligated with cloning vector pBsk(+).The recombinant clones have been obtained from the selected medium. The results of DNA sequences analysis show that the nucleotide sequences of ORF share more than 85% homologies as compared with those of calmodulin genes of several other plants.Similar to rice and apple, the ORF is interrupted by an intron behind the 75th nucleotide.展开更多
Septic encephalopathy is a frequent complication of sepsis,but there are few studies examining the role of micro RNAs（mi Rs） in its pathogenesis.In this study,a mi R-219 mimic was transfected into rat hippocampal ne...Septic encephalopathy is a frequent complication of sepsis,but there are few studies examining the role of micro RNAs（mi Rs） in its pathogenesis.In this study,a mi R-219 mimic was transfected into rat hippocampal neurons to model mi R-219 overexpression.A protective effect of mi R-219 was observed for glutamate-induced neurotoxicity of rat hippocampal neurons,and an underlying mechanism involving calmodulin-dependent protein kinase II γ（Ca MKIIγ） was demonstrated.mi R-219 and Ca MKIIγ m RNA expression induced by glutamate in hippocampal neurons was determined by quantitative real-time reverse transcription-polymerase chain reaction（q RT-PCR）.After neurons were transfected with mi R-219 mimic,effects on cell viability and apoptosis were measured by 3-（4,5-dimethylthiazolyl-2）-2,5-diphenyltetrazolium bromide（MTT） assay and flow cytometry.In addition,a luciferase reporter gene system was used to confirm Ca MKIIγ as a target gene of mi R-219.Western blot assay and rescue experiments were also utilized to detect Ca MKIIγ expression and further verify that mi R-219 in hippocampal neurons exerted its effect through regulation of Ca MKIIγ.MTT assay and q RT-PCR results revealed obvious decreases in cell viability and mi R-219 expression after glutamate stimulation,while Ca MKIIγ m RNA expression was increased.MTT,flow cytometry,and caspase-3 activity assays showed that mi R-219 overexpression could elevate glutamate-induced cell viability,and reduce cell apoptosis and caspase-3 activity.Moreover,luciferase Ca MKIIγ-reporter activity was remarkably decreased by co-transfection with mi R-219 mimic,and the results of a rescue experiment showed that Ca MKIIγ overexpression could reverse the biological effects of mi R-219.Collectively,these findings verify that mi R-219 expression was decreased in glutamate-induced neurons,Ca MKIIγ was a target gene of mi R-219,and mi R-219 alleviated glutamate-induced neuronal excitotoxicity by negatively controlling Ca MKIIγ expression.展开更多
A rice CaMBP gene, OsCaMBP （AB363406）, was isolated from a chilling treated rice using the fluorescent differential display （FDD） screening method. Its cDNA sequence （2094 bp） contains an opening reading frame ...A rice CaMBP gene, OsCaMBP （AB363406）, was isolated from a chilling treated rice using the fluorescent differential display （FDD） screening method. Its cDNA sequence （2094 bp） contains an opening reading frame （ORF） encoding a 569 amino acids protein （63.2 kD）. OsCaMBP has the typical structural features of the CaMBP family, including the conserved IQ calmodulin-binding motif at the N-terminus. Homology analysis revealed 38.25%-47.28% identities of OsCaMBP with other CaMBPs in plants. RT-PCR analysis showed that the expression of OsCaMBP was remarkably inducible under the chilling （8℃） and heat-shock （42℃） treatments. OsCaMBP was undetectable under the normal conditions, and induced under the chilling treatment for 1 h, as well as the heat-shock treatment for 15 min, suggesting that the gene plays important roles in the signaling pathway in rice under both chilling and heat-shock stresses.展开更多
为分析黑鲷♀×真鲷♂(Acanthopagrus schlegelii♀×Pagrosomus major♂)反交子代在生长、发育等经济性状优于黑鲷亲本(A.schlegelii)的分子遗传差异,本研究克隆了反交子代(PA)与黑鲷(As)的Ca M基因,运用生物信息学方法对基因P...为分析黑鲷♀×真鲷♂(Acanthopagrus schlegelii♀×Pagrosomus major♂)反交子代在生长、发育等经济性状优于黑鲷亲本(A.schlegelii)的分子遗传差异,本研究克隆了反交子代(PA)与黑鲷(As)的Ca M基因,运用生物信息学方法对基因PACa M与As Ca M的序列进行了详细分析;同时通过荧光定量分析了PACa M与As Ca M在仔鱼及2龄成鱼不同组织的表达特征。研究结果表明,PACa M基因c DNA全长1180 bp,As Ca M基因c DNA全长1241 bp,均具有一个450 bp的开放阅读框,编码149个氨基酸,分子量约16.84 k D,等电点为4.09;序列比对、结构比较等分析表明,PACa M和As Ca M属Ca M基因家族,具有4个EF-hand钙结合功能域。定量分析表明Ca M在两种鱼的仔鱼及成鱼的脑与性腺中有较高表达;PACa M和As Ca M在仔鱼及成鱼的鳃、肌肉及性腺中的表达存在显著差异(P<0.05),在脑、肝及肾中的表达没有明显差异(P>0.05);PACa M在仔鱼中表达量最高,As Ca M在成鱼性腺中表达最高,均显示了Ca M基因在生长与繁殖中的重要作用。研究结果为鲷科属间杂交获得的子代与亲本性状差异的功能基因表达提供一些基础资料。展开更多
基金This research was funded by the Natural Science Foundation of Fujian Province,China(2018J01470 and 2021J01137)Scientific research projects of introducing talents in Wuyi University(YJ202109)+2 种基金Special fund for scientific and technological innovation of Fujian Agriculture and Forestry University(CXZX2020081A)China Agriculture Research System of MOF and MARA(CARS-17)The funders had no role in the study design,data collection,and analysis,decision to publish,or preparation of the manuscript.
文摘Calmodulin(CaM)proteins play a key role in signal transduction under various stresses.In the present study,the effects of a sugarcane ScCaM gene(NCBI accession number:GQ246454)on drought and salt stress tolerance in transgenic Arabidopsis thaliana and Escherichia coli cells were evaluated.The results demonstrated a significant negative role of ScCaM in the drought and salt stress tolerance of transgenic lines of A.thaliana,as indicated by the phenotypes.In addition,the expression of AtP5CS and AtRD29A,two genes tightly related to stress resistance,was significantly lower in the overexpression lines than in the wild type.The growth of E.coli BL21 cells expressing ScCaM showed weaker tolerance under mannitol and NaCl stress.Taken together,this study revealed that the ScCaM gene plays a negative regulatory role in both mannitol and NaCl stresses,and it possibly exerts protective mechanisms common in both prokaryotes and eukaryotes under stress conditions.
文摘We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 cells. After treatment with 1 mM db-cAMP for 1 hour and 2 hours, there was an early and repldly reduced in gene expression of Calmodulin and c-fos respectively. After db-cAMP treatment for 4 -5 days, the number of capping cells of ConA binding decreased significantly and the cell surface microvllll decreased as well. The growth of treated cells was inhibited markedly. By using 4F1 cDNA probe, which is preferentially expressed In G1 phase, we have found that the db- cAMP treated cells were accumulated at G1 phase. Of particular interest is the fact that the distribution of microtubules, mlcrofilaments and fibronectln were recovered after treatment with 1 mM db-cAMP for 6 days. It is suggested that the Inhibition of proliferation, alteration, of phenotype and reco- very of cytoskeleton is transformed cells after treatment with db-cAMP are related to the Inhibition of gene expression of Calmodulin.
基金Supported by the national science and technology ministry plant transgenic and industrialization foundation of China (J00-B-14) the foundation of Key Laboratory of halobios engineering SOA
文摘Calmodulin is a calcium binding protein that modulates the activity of diverse groups of protein including some protein kinase, adenylate cyclases and ATPase. Here we use the total DNA of Sonneratia paracaseolaris as the template of the polymerase chain reaction (PCR). The PCR primers have been designed and synthesized according to the 5-and 3-terminal oligonucleotide sequences of Calmodulin gene of plants in Genbank and ligated with cloning vector pBsk(+).The recombinant clones have been obtained from the selected medium. The results of DNA sequences analysis show that the nucleotide sequences of ORF share more than 85% homologies as compared with those of calmodulin genes of several other plants.Similar to rice and apple, the ORF is interrupted by an intron behind the 75th nucleotide.
基金supported by the National Natural Science Foundation of China,No.81101159the Natural Science Foundation of Jiangsu Province of China,No.BK20151268
文摘Septic encephalopathy is a frequent complication of sepsis,but there are few studies examining the role of micro RNAs（mi Rs） in its pathogenesis.In this study,a mi R-219 mimic was transfected into rat hippocampal neurons to model mi R-219 overexpression.A protective effect of mi R-219 was observed for glutamate-induced neurotoxicity of rat hippocampal neurons,and an underlying mechanism involving calmodulin-dependent protein kinase II γ（Ca MKIIγ） was demonstrated.mi R-219 and Ca MKIIγ m RNA expression induced by glutamate in hippocampal neurons was determined by quantitative real-time reverse transcription-polymerase chain reaction（q RT-PCR）.After neurons were transfected with mi R-219 mimic,effects on cell viability and apoptosis were measured by 3-（4,5-dimethylthiazolyl-2）-2,5-diphenyltetrazolium bromide（MTT） assay and flow cytometry.In addition,a luciferase reporter gene system was used to confirm Ca MKIIγ as a target gene of mi R-219.Western blot assay and rescue experiments were also utilized to detect Ca MKIIγ expression and further verify that mi R-219 in hippocampal neurons exerted its effect through regulation of Ca MKIIγ.MTT assay and q RT-PCR results revealed obvious decreases in cell viability and mi R-219 expression after glutamate stimulation,while Ca MKIIγ m RNA expression was increased.MTT,flow cytometry,and caspase-3 activity assays showed that mi R-219 overexpression could elevate glutamate-induced cell viability,and reduce cell apoptosis and caspase-3 activity.Moreover,luciferase Ca MKIIγ-reporter activity was remarkably decreased by co-transfection with mi R-219 mimic,and the results of a rescue experiment showed that Ca MKIIγ overexpression could reverse the biological effects of mi R-219.Collectively,these findings verify that mi R-219 expression was decreased in glutamate-induced neurons,Ca MKIIγ was a target gene of mi R-219,and mi R-219 alleviated glutamate-induced neuronal excitotoxicity by negatively controlling Ca MKIIγ expression.
基金supported by the Program for Changjiang Scholars and Innovative Research Team in University of China (Grant No. IRT0453)the National Science & Technology Pillar Program of China in the Eleventh Five-Year Plan Period (Grant No. 2007BAD81B00).
文摘A rice CaMBP gene, OsCaMBP （AB363406）, was isolated from a chilling treated rice using the fluorescent differential display （FDD） screening method. Its cDNA sequence （2094 bp） contains an opening reading frame （ORF） encoding a 569 amino acids protein （63.2 kD）. OsCaMBP has the typical structural features of the CaMBP family, including the conserved IQ calmodulin-binding motif at the N-terminus. Homology analysis revealed 38.25%-47.28% identities of OsCaMBP with other CaMBPs in plants. RT-PCR analysis showed that the expression of OsCaMBP was remarkably inducible under the chilling （8℃） and heat-shock （42℃） treatments. OsCaMBP was undetectable under the normal conditions, and induced under the chilling treatment for 1 h, as well as the heat-shock treatment for 15 min, suggesting that the gene plays important roles in the signaling pathway in rice under both chilling and heat-shock stresses.
文摘为分析黑鲷♀×真鲷♂(Acanthopagrus schlegelii♀×Pagrosomus major♂)反交子代在生长、发育等经济性状优于黑鲷亲本(A.schlegelii)的分子遗传差异,本研究克隆了反交子代(PA)与黑鲷(As)的Ca M基因,运用生物信息学方法对基因PACa M与As Ca M的序列进行了详细分析;同时通过荧光定量分析了PACa M与As Ca M在仔鱼及2龄成鱼不同组织的表达特征。研究结果表明,PACa M基因c DNA全长1180 bp,As Ca M基因c DNA全长1241 bp,均具有一个450 bp的开放阅读框,编码149个氨基酸,分子量约16.84 k D,等电点为4.09;序列比对、结构比较等分析表明,PACa M和As Ca M属Ca M基因家族,具有4个EF-hand钙结合功能域。定量分析表明Ca M在两种鱼的仔鱼及成鱼的脑与性腺中有较高表达;PACa M和As Ca M在仔鱼及成鱼的鳃、肌肉及性腺中的表达存在显著差异(P<0.05),在脑、肝及肾中的表达没有明显差异(P>0.05);PACa M在仔鱼中表达量最高,As Ca M在成鱼性腺中表达最高,均显示了Ca M基因在生长与繁殖中的重要作用。研究结果为鲷科属间杂交获得的子代与亲本性状差异的功能基因表达提供一些基础资料。
