[ Objective] This study aimed to optimize the cedon of antimicrobial peptide SMAP-29 gene and express SMAP-29 in Pichia pastoris. [ Method] Ac- cording to the published amino acid sequence of SMAP-29, a gene encoding ...[ Objective] This study aimed to optimize the cedon of antimicrobial peptide SMAP-29 gene and express SMAP-29 in Pichia pastoris. [ Method] Ac- cording to the published amino acid sequence of SMAP-29, a gene encoding SMAP-29 （sheep myeloid antibacterial peptides-29） mature peptide was synthesized and was biased codon usage of Pichia pastoris. The synthesized gene was inserted into Pichia pastoris expression vector pPIC3.5K, transformed into Pichia pastoris strain GSll5 by electmporation, and screened with G418 for high-copy recombinants. The methanol utility plus phenotype （Mut ＋ ） was selected with MM, MD and PCR. The correctly constructed recombinant strain was induced with methanol. Expression of SMAP-29 was analyzed by lysing the induced yeast cells with Tricine- SDS-PAGE. [ Result] After induced with methanol for 2 d, a 3.2 kD induced band was detected in the cell lysate which was consistent to the predicted molecular weight of SMAP-29 ; the expression products purified with gel filtration chromatography showed significant antibacterial effect against Staphyloccocus aureus and Mo- nilia albican but no significant antibacterial effect against Escherichia coli. [ Conclusion] This study laid the foundation for the application of SMAP-29 in biomedi- cine, agriculture and other fields.展开更多
Developers still need design workflow system according to users＇ specific needs, though workflow management coalition standardized the five kinds of abstract interfaces in workflow reference model. Specific business ...Developers still need design workflow system according to users＇ specific needs, though workflow management coalition standardized the five kinds of abstract interfaces in workflow reference model. Specific business process characteristics are still supported by specific workflow system. A set of common functionalities of workflow engine are abstracted from business component, so the reusability of business component is extended into workflow engine and composition method is proposed. Needs of different business requirements and characteristics are met by reusing the workflow engine.展开更多
This study aimed to identify the potential metabolic biomarkers of the Badagan constitution(BC) in Mongolian medicine. Serum samples from the participants with BCs(n = 32) and aggregative constitutions(n = 30) were an...This study aimed to identify the potential metabolic biomarkers of the Badagan constitution(BC) in Mongolian medicine. Serum samples from the participants with BCs(n = 32) and aggregative constitutions(n = 30) were analyzed by using ultrahigh-performance liquid chromatography/mass spectrometry. Meanwhile, principal component analysis and orthogonal partial least squares discriminant analysis were used to characterize the endogenous metabolites and potential biomarkers, respectively. Fifteen of the 18 biomarkers in six metabolic pathways were significantly upregulated,including phosphatidylserine, sphingolipids, tryptophan, riboflavin and glutathione, and three biomarkers were significantly downregulated,including lysophosphatidylcholine(LysoPC)(18:1), LysoPC(16:1), and lysophosphatidylethanolamine(LysoPE)(22:2). This study also implied that sphingolipid metabolism, glycerophospholipid metabolism, and tryptophan metabolism played important roles in the BC. Therefore, metabolomics may improve the diagnostic efficacy of the BC in terms of the accuracy and comprehensiveness of a diagnosis based on this constitution. This result further reveals the mechanism of the constitution type in Mongolian medicine and provides a reference for the treatment of related diseases.展开更多
基金Supported by Basic Research Project from Science and Technology Bureau of Xinjiang Production and Construction Corps (2006JC01)National Natural Science Foundation of China (31160218)
文摘[ Objective] This study aimed to optimize the cedon of antimicrobial peptide SMAP-29 gene and express SMAP-29 in Pichia pastoris. [ Method] Ac- cording to the published amino acid sequence of SMAP-29, a gene encoding SMAP-29 （sheep myeloid antibacterial peptides-29） mature peptide was synthesized and was biased codon usage of Pichia pastoris. The synthesized gene was inserted into Pichia pastoris expression vector pPIC3.5K, transformed into Pichia pastoris strain GSll5 by electmporation, and screened with G418 for high-copy recombinants. The methanol utility plus phenotype （Mut ＋ ） was selected with MM, MD and PCR. The correctly constructed recombinant strain was induced with methanol. Expression of SMAP-29 was analyzed by lysing the induced yeast cells with Tricine- SDS-PAGE. [ Result] After induced with methanol for 2 d, a 3.2 kD induced band was detected in the cell lysate which was consistent to the predicted molecular weight of SMAP-29 ; the expression products purified with gel filtration chromatography showed significant antibacterial effect against Staphyloccocus aureus and Mo- nilia albican but no significant antibacterial effect against Escherichia coli. [ Conclusion] This study laid the foundation for the application of SMAP-29 in biomedi- cine, agriculture and other fields.
文摘Developers still need design workflow system according to users＇ specific needs, though workflow management coalition standardized the five kinds of abstract interfaces in workflow reference model. Specific business process characteristics are still supported by specific workflow system. A set of common functionalities of workflow engine are abstracted from business component, so the reusability of business component is extended into workflow engine and composition method is proposed. Needs of different business requirements and characteristics are met by reusing the workflow engine.
基金supported by the National Natural Science Foundation of China(grant number 81660828)the Open Fund Project of Engineering Technology Research Center of Mongolian Medicine of Inner Mongolia(grant numbers MDK2018070,MDK2019034)+2 种基金MDK2019036National and local Joint Engineering Research Center for Mongolian Medicine Research Open Fund Projects(MDK2021035)the Natural Science Foundation of Inner Mongolia of China(grant number 2019MS08040)。
文摘This study aimed to identify the potential metabolic biomarkers of the Badagan constitution(BC) in Mongolian medicine. Serum samples from the participants with BCs(n = 32) and aggregative constitutions(n = 30) were analyzed by using ultrahigh-performance liquid chromatography/mass spectrometry. Meanwhile, principal component analysis and orthogonal partial least squares discriminant analysis were used to characterize the endogenous metabolites and potential biomarkers, respectively. Fifteen of the 18 biomarkers in six metabolic pathways were significantly upregulated,including phosphatidylserine, sphingolipids, tryptophan, riboflavin and glutathione, and three biomarkers were significantly downregulated,including lysophosphatidylcholine(LysoPC)(18:1), LysoPC(16:1), and lysophosphatidylethanolamine(LysoPE)(22:2). This study also implied that sphingolipid metabolism, glycerophospholipid metabolism, and tryptophan metabolism played important roles in the BC. Therefore, metabolomics may improve the diagnostic efficacy of the BC in terms of the accuracy and comprehensiveness of a diagnosis based on this constitution. This result further reveals the mechanism of the constitution type in Mongolian medicine and provides a reference for the treatment of related diseases.
