AdoMet plays numerous roles of being the major methyl-group donor in trans-methylation reactions. To gain insight into the possible functions of the AdoMet protein of Suaeda salsa L. in response to salt stress, S aden...AdoMet plays numerous roles of being the major methyl-group donor in trans-methylation reactions. To gain insight into the possible functions of the AdoMet protein of Suaeda salsa L. in response to salt stress, S adenosylmethionine synthetase gene (SAMS2) was analyzed. We isolated SAMS2 cDNA clone (AF321001) from a lambda -Zap cDNA library constructed from the halophyte S. salsa Pall aerial tissue treated with 400 mmol/L NaCl. SsSAMS2 was found to encode a S-adenolyl-L-methionine synthetase enzyme (AdoMet synthetase). The fragment was 1 531 bp with an open reading frame of 395 amino acids, the calculated molecular weight was about 43 kD. SsSAMS2 showed the highest homology to SAMS2 gene of Catharanthus roseus G. Don., with 93% identity in deduced amino acid sequence. Southern blotting analysis showed that SsSAMS2 might be a two-copy gene in S. salsa genome. Northern blot indicated that the cDNA was up-regulated by salt and other stresses. Enzyme activity assay indicated that the activity of SAMS2 increased under NaCl stress.展开更多
[Objective] The mRNA expression level changes of S-adenosylmethionine synthetase （SAMS） under low temperature stress was studied. [Method] Total RNA were extracted from leaves, stem and earthnut of sweet potato 0,12...[Objective] The mRNA expression level changes of S-adenosylmethionine synthetase （SAMS） under low temperature stress was studied. [Method] Total RNA were extracted from leaves, stem and earthnut of sweet potato 0,12,24,48 and 72 h after low temperature treatement, mRNA expression level was analyzed by reverse expression and Real-time PCR technique. [Result] The expression quality of the gene extracted from leaves, stem and earthnut increased and the expression quality reached the peak point 24,72 and 72 h after low temperature treatment respectively. The expression change of earthnut was the biggest. [Conclusion] Low temperature was good for increasing mRNA expression of relevart genes.展开更多
[Objective] The research aimed to study the secreted expression of S-edenosyl-L-methionine synthetase （SAMS） in Pichia pastoris. Method ] The gene coding SAMS, from the genomic DNA of Saccharomyces cerevisiae, was ...[Objective] The research aimed to study the secreted expression of S-edenosyl-L-methionine synthetase （SAMS） in Pichia pastoris. Method ] The gene coding SAMS, from the genomic DNA of Saccharomyces cerevisiae, was amplified by PCR and inserted into the secreted expression vector pPIC9K to get recombinant plasmid. The recombinant plasmid pPIC9K-sarr~ was integrated into Pichia pastoris GSl15 genome by electroporation and induced by methanol. The activity of the recombinant enzyme was measured using high-pedormance liquid chroma- tography （HPLC） by determining the production of S-adenosy-L-methionine （SAM） with the enzyme secreted. [ ResultJ The molecular weight of the expression protein identified by SDS-PAGE was about 50 kD, being larger than the theoretical molecular mass of SAMS, which might be due to the glycosytation in the process of secretion. Methanol-induction as well as preliminary purification could enhance the enzyme activity, espe- cially the latter, after which the specific activity of SAMS was improved to 61.48 U/rng. [Conclusion] SAMS with biological activity was secreted successfully in Pichia pastoris GSl15 for the first time. And it is the start for the genetic engineering strains to open up prospects for industrial production.展开更多
The leaching kinetics of copper from low-grade copper ore was investigated in ammonia-ammonium sulfate solution with sodium persulfate. The effect parameters of stirring speed, temperature, particle size, concentratio...The leaching kinetics of copper from low-grade copper ore was investigated in ammonia-ammonium sulfate solution with sodium persulfate. The effect parameters of stirring speed, temperature, particle size, concentrations of ammonia, ammonium sulfate and sodium persulfate were determined. The results show that the leaching rate is nearly independent of agitation above 300 r/min and increases with the increase of temperature, concentrations of ammonia, ammonium sulfate and sodium persulfate. The EDS analysis and phase quantitative analysis of the residues indicate that bornite can be dissolved by persulfate oxidization. The leaching kinetics with activation energy of 22.91 kJ/mol was analyzed by using a new shrinking core model （SCM） in which both the interfacial transfer and diffusion across the product layer affect the leaching rate. A semi-empirical rate equation was obtained to describe the leaching process and the empirical reaction orders with respect to the concentrations of ammonia, ammonium sulfate and sodium persulfate are 0.5, 1.2 and 0.5, respectively.展开更多
A variant cell line of Medicago saliva L. resistant to methionine (100 mmol/L) was selected .from calli treated with 1.5 mmol/L sodium azide. Plants were regenerated from this selected cell line. After growing for 6 m...A variant cell line of Medicago saliva L. resistant to methionine (100 mmol/L) was selected .from calli treated with 1.5 mmol/L sodium azide. Plants were regenerated from this selected cell line. After growing for 6 months on the medium devoid of selection pressure, the variant cell line still showed high level of resistance, which was 7.2-fold higher than that of the wild type. This indicated the stability of the resistance expression. The variant cell line also expressed a 3.3-fold higher cross-resistance to ethionine, than that of the wild type. The contents of methionine, lysine, threonine and isoleucine in the variant calli were respectively 2.0, 1.43, 1.71 and 1.57 times higher than those in the wild type. However, in the variant regenerated plantlets they respectively were 2.0, 1.33, 1.22 and 1.22 times higher than those in the wild type regenerants. Four new protein bands, viz. 65 kD, 23 kD, 20 kD and 18 kD, appeared in the SDS-PAGE pattern of soluble proteins in the variant calli. In the peroxidase isoenzyme electrophoresis pattern, two new bands were also observed in the variant calli.展开更多
文摘AdoMet plays numerous roles of being the major methyl-group donor in trans-methylation reactions. To gain insight into the possible functions of the AdoMet protein of Suaeda salsa L. in response to salt stress, S adenosylmethionine synthetase gene (SAMS2) was analyzed. We isolated SAMS2 cDNA clone (AF321001) from a lambda -Zap cDNA library constructed from the halophyte S. salsa Pall aerial tissue treated with 400 mmol/L NaCl. SsSAMS2 was found to encode a S-adenolyl-L-methionine synthetase enzyme (AdoMet synthetase). The fragment was 1 531 bp with an open reading frame of 395 amino acids, the calculated molecular weight was about 43 kD. SsSAMS2 showed the highest homology to SAMS2 gene of Catharanthus roseus G. Don., with 93% identity in deduced amino acid sequence. Southern blotting analysis showed that SsSAMS2 might be a two-copy gene in S. salsa genome. Northern blot indicated that the cDNA was up-regulated by salt and other stresses. Enzyme activity assay indicated that the activity of SAMS2 increased under NaCl stress.
文摘[Objective] The mRNA expression level changes of S-adenosylmethionine synthetase （SAMS） under low temperature stress was studied. [Method] Total RNA were extracted from leaves, stem and earthnut of sweet potato 0,12,24,48 and 72 h after low temperature treatement, mRNA expression level was analyzed by reverse expression and Real-time PCR technique. [Result] The expression quality of the gene extracted from leaves, stem and earthnut increased and the expression quality reached the peak point 24,72 and 72 h after low temperature treatment respectively. The expression change of earthnut was the biggest. [Conclusion] Low temperature was good for increasing mRNA expression of relevart genes.
文摘[Objective] The research aimed to study the secreted expression of S-edenosyl-L-methionine synthetase （SAMS） in Pichia pastoris. Method ] The gene coding SAMS, from the genomic DNA of Saccharomyces cerevisiae, was amplified by PCR and inserted into the secreted expression vector pPIC9K to get recombinant plasmid. The recombinant plasmid pPIC9K-sarr~ was integrated into Pichia pastoris GSl15 genome by electroporation and induced by methanol. The activity of the recombinant enzyme was measured using high-pedormance liquid chroma- tography （HPLC） by determining the production of S-adenosy-L-methionine （SAM） with the enzyme secreted. [ ResultJ The molecular weight of the expression protein identified by SDS-PAGE was about 50 kD, being larger than the theoretical molecular mass of SAMS, which might be due to the glycosytation in the process of secretion. Methanol-induction as well as preliminary purification could enhance the enzyme activity, espe- cially the latter, after which the specific activity of SAMS was improved to 61.48 U/rng. [Conclusion] SAMS with biological activity was secreted successfully in Pichia pastoris GSl15 for the first time. And it is the start for the genetic engineering strains to open up prospects for industrial production.
基金Project(2007CB613601)supported by the National Basic Research Program of ChinaProject(10C1095)supported by the Foundation of Hunan Educational Committee,China
文摘The leaching kinetics of copper from low-grade copper ore was investigated in ammonia-ammonium sulfate solution with sodium persulfate. The effect parameters of stirring speed, temperature, particle size, concentrations of ammonia, ammonium sulfate and sodium persulfate were determined. The results show that the leaching rate is nearly independent of agitation above 300 r/min and increases with the increase of temperature, concentrations of ammonia, ammonium sulfate and sodium persulfate. The EDS analysis and phase quantitative analysis of the residues indicate that bornite can be dissolved by persulfate oxidization. The leaching kinetics with activation energy of 22.91 kJ/mol was analyzed by using a new shrinking core model （SCM） in which both the interfacial transfer and diffusion across the product layer affect the leaching rate. A semi-empirical rate equation was obtained to describe the leaching process and the empirical reaction orders with respect to the concentrations of ammonia, ammonium sulfate and sodium persulfate are 0.5, 1.2 and 0.5, respectively.
基金This research project was supported by the National Natural Science Foundation of China.
文摘A variant cell line of Medicago saliva L. resistant to methionine (100 mmol/L) was selected .from calli treated with 1.5 mmol/L sodium azide. Plants were regenerated from this selected cell line. After growing for 6 months on the medium devoid of selection pressure, the variant cell line still showed high level of resistance, which was 7.2-fold higher than that of the wild type. This indicated the stability of the resistance expression. The variant cell line also expressed a 3.3-fold higher cross-resistance to ethionine, than that of the wild type. The contents of methionine, lysine, threonine and isoleucine in the variant calli were respectively 2.0, 1.43, 1.71 and 1.57 times higher than those in the wild type. However, in the variant regenerated plantlets they respectively were 2.0, 1.33, 1.22 and 1.22 times higher than those in the wild type regenerants. Four new protein bands, viz. 65 kD, 23 kD, 20 kD and 18 kD, appeared in the SDS-PAGE pattern of soluble proteins in the variant calli. In the peroxidase isoenzyme electrophoresis pattern, two new bands were also observed in the variant calli.
