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
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
INTRODUCTIONCalmodulin（CaM）,widely distributed in almost alleukaryotic cells,is a major intracellular calcium receptorresponsible for mediating the Ca<sup>2+</sup> signal to a multitude ofdifferent enzym...INTRODUCTIONCalmodulin（CaM）,widely distributed in almost alleukaryotic cells,is a major intracellular calcium receptorresponsible for mediating the Ca<sup>2+</sup> signal to a multitude ofdifferent enzyme systems and is thought to play a vital rolein the regulation of cell proliferative cycle.Recently,展开更多
Sharp eyespot,mainly caused by the soil-borne fungus Rhizoctonia cerealis,affects wheat(Triticum aestivum L.)production worldwide.In this study,we isolated TaCML36 gene encoding a wheat calmodulin-like protein,and stu...Sharp eyespot,mainly caused by the soil-borne fungus Rhizoctonia cerealis,affects wheat(Triticum aestivum L.)production worldwide.In this study,we isolated TaCML36 gene encoding a wheat calmodulin-like protein,and studied its defense role in protection against R.cerealis.Transcription of TaCML36 was significantly elevated by both R.cerealis infection and exogenous ethylene treatment.Transcription was higher in resistant wheat lines than in susceptible ones.There were copies of TaCML36 on chromosomes 5A,5B,and 5D.The TaCML36 protein is composed of 183 amino acids and contains two calcium-binding EFhand domains.Subcellular localization assays in wheat indicated that TaCML36 localizes in both the cytoplasm and nucleus.Virus-induced gene silencing and disease assessment indicated that compared to the controls,TaCML36-silenced wheat plants displayed significantly reduced resistance to R.cerealis and had greater fungal biomass,suggesting that knockdown of TaCML36 impaired host resistance.Knockdown of TaCML36 also significantly repressed expression of pathogenesis-related genes such as Chitinase 1,PDF35,and PR17C,the ethylene response factor-encoding gene TaPIE1,and ethylene biosynthesis gene ACO2.Collectively,our results suggest that TaCML36 positively participates in the innate immune response to R.cerealis infection by modulating expression of defense-associated genes possibly in the ethylene signaling pathway.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
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
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
The treatment of wheat （ Triticum aestivum L.） seedlings with CaCl 2 increased mRNA levels of hsp26 gene at 37 ℃ for 1 h, while that with Ca 2+ chelator EGTA decreased the expression of the hsp26 gene. The expressi...The treatment of wheat （ Triticum aestivum L.） seedlings with CaCl 2 increased mRNA levels of hsp26 gene at 37 ℃ for 1 h, while that with Ca 2+ chelator EGTA decreased the expression of the hsp26 gene. The expression of the wheat calmodulin gene CaM12 is rapidly upregulated in wheat seedlings by heat shock at 37 ℃. The heatinduced upregulation of CaM12 occurred earlier than that of the hsp26 gene did. Calmodulin antagonist N6aminohexyl5chloro1naphthalenesulfonamid （W7） inhibited the expression of the hsp26 gene in wheat seedlings at 37 ℃ for 1 h. These results implied that the calciumcalmodulin is probably involved in the signal transduction of the hsp26 gene expression induced by heat shock.展开更多
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.
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the Natural Science Fundation of Jiangsu Province,№BK95141307
文摘INTRODUCTIONCalmodulin（CaM）,widely distributed in almost alleukaryotic cells,is a major intracellular calcium receptorresponsible for mediating the Ca<sup>2+</sup> signal to a multitude ofdifferent enzyme systems and is thought to play a vital rolein the regulation of cell proliferative cycle.Recently,
基金funded by the National “Key Sci-Tech” Project (2016ZX08002-001-004)
文摘Sharp eyespot,mainly caused by the soil-borne fungus Rhizoctonia cerealis,affects wheat(Triticum aestivum L.)production worldwide.In this study,we isolated TaCML36 gene encoding a wheat calmodulin-like protein,and studied its defense role in protection against R.cerealis.Transcription of TaCML36 was significantly elevated by both R.cerealis infection and exogenous ethylene treatment.Transcription was higher in resistant wheat lines than in susceptible ones.There were copies of TaCML36 on chromosomes 5A,5B,and 5D.The TaCML36 protein is composed of 183 amino acids and contains two calcium-binding EFhand domains.Subcellular localization assays in wheat indicated that TaCML36 localizes in both the cytoplasm and nucleus.Virus-induced gene silencing and disease assessment indicated that compared to the controls,TaCML36-silenced wheat plants displayed significantly reduced resistance to R.cerealis and had greater fungal biomass,suggesting that knockdown of TaCML36 impaired host resistance.Knockdown of TaCML36 also significantly repressed expression of pathogenesis-related genes such as Chitinase 1,PDF35,and PR17C,the ethylene response factor-encoding gene TaPIE1,and ethylene biosynthesis gene ACO2.Collectively,our results suggest that TaCML36 positively participates in the innate immune response to R.cerealis infection by modulating expression of defense-associated genes possibly in the ethylene signaling pathway.
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.
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.
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。
文摘The treatment of wheat （ Triticum aestivum L.） seedlings with CaCl 2 increased mRNA levels of hsp26 gene at 37 ℃ for 1 h, while that with Ca 2+ chelator EGTA decreased the expression of the hsp26 gene. The expression of the wheat calmodulin gene CaM12 is rapidly upregulated in wheat seedlings by heat shock at 37 ℃. The heatinduced upregulation of CaM12 occurred earlier than that of the hsp26 gene did. Calmodulin antagonist N6aminohexyl5chloro1naphthalenesulfonamid （W7） inhibited the expression of the hsp26 gene in wheat seedlings at 37 ℃ for 1 h. These results implied that the calciumcalmodulin is probably involved in the signal transduction of the hsp26 gene expression induced by heat shock.
