[Objectives]The paper was to analyze and predict the structure and characteristics of xylanase xynMF13-GH10 gene and its encoded protein.[Methods]xynMF13-GH10 gene was predicted by NCBI and various information analysi...[Objectives]The paper was to analyze and predict the structure and characteristics of xylanase xynMF13-GH10 gene and its encoded protein.[Methods]xynMF13-GH10 gene was predicted by NCBI and various information analysis tools in ExPASy website,as well as SignaIP 5.0,DNAman,TMHMM,SOPMA and SWISS-Model,and the characteristics and functions of protein structure encoded by the gene were predicted.[Results]The gene is 1332 bp in length,the coding region is 1-1332 bp,and the gene encodes 443 amino acids.The xynMF13-GH10 gene has high homology with xylanase in many species,and it has the highest homology with Paraphaeosphaeria sporulosa endoxylanase-like protein,with the consistency reaching 78.91%and e-value reaching 2e-159.The secondary structure consists of 48.31%random curl,30.25%α-helix structure,16.70%extended chain and 4.74%β-corner.[Conclusions]The results provide a reference for revealing the physiological function and expression regulation mechanism of xynMF13-GH10 gene in the future.展开更多
The Aspergillus niger XynB gene and core promoter region of porcine RELMβgene were cloned into pcDNA3.1(-),and an intestine-specific expression vector pcDNA3.1-RELMβ-XynB-Myc-GFP carrying green fluorescence and Myc ...The Aspergillus niger XynB gene and core promoter region of porcine RELMβgene were cloned into pcDNA3.1(-),and an intestine-specific expression vector pcDNA3.1-RELMβ-XynB-Myc-GFP carrying green fluorescence and Myc double tags was constructed.The vector was transfected into human colon cancer cells(HT29)and human liver cancer cells(Bel7402)using liposomes.Fluorescence microscopy revealed that the vector could specifically express green fluorescent protein(GFP)in HT29 cells.RT-PCR and Western Blot were performed on the HT29 cells transfected with the expression vector,and the results showed that the XynB gene was normally transcribed in HT29 cells,and the target protein expression was detected in the cells.展开更多
Extracellular xylanase XYNB from Streptomy-ces olivaceoviridis A1 has been purified and characterized. The optimal pH value and temperature of XYNB for its ac-tivity are 5.2 and 60℃, respectively. The specific activi...Extracellular xylanase XYNB from Streptomy-ces olivaceoviridis A1 has been purified and characterized. The optimal pH value and temperature of XYNB for its ac-tivity are 5.2 and 60℃, respectively. The specific activity of XYNB is as high as 2869.78 U/mg. Metal cations, EDTA and SDS have no effects on enzyme activity of XYNB. The gene xynB coding mature protein of XYNB has been cloned by PCR. The forward oligonucleotide primer used in the PCR reaction was synthesized based on the N-terminal amino acid sequence of XYNB mature protein, and the reverse oligonu-cleotide primers are random oligonucleotide. The cloned gene xynB is 576 bp long and its G+C content is 64.3%. The xynB encodes 191 amino acid residues, and the putative mo-lecular weight of XYNB is 20.839 kD. The xynB has been expressed in E. coli, and the expressed xylanase has normal bioactivity.展开更多
基金Public Service Platform Project for Industrial Chain Collaborative Innovation(ZHC 2017CB82)Public Service Platform Subproject for Research and Development of Marine Biomedical Resources in the South China Sea(XM-202008-01B2).
文摘[Objectives]The paper was to analyze and predict the structure and characteristics of xylanase xynMF13-GH10 gene and its encoded protein.[Methods]xynMF13-GH10 gene was predicted by NCBI and various information analysis tools in ExPASy website,as well as SignaIP 5.0,DNAman,TMHMM,SOPMA and SWISS-Model,and the characteristics and functions of protein structure encoded by the gene were predicted.[Results]The gene is 1332 bp in length,the coding region is 1-1332 bp,and the gene encodes 443 amino acids.The xynMF13-GH10 gene has high homology with xylanase in many species,and it has the highest homology with Paraphaeosphaeria sporulosa endoxylanase-like protein,with the consistency reaching 78.91%and e-value reaching 2e-159.The secondary structure consists of 48.31%random curl,30.25%α-helix structure,16.70%extended chain and 4.74%β-corner.[Conclusions]The results provide a reference for revealing the physiological function and expression regulation mechanism of xynMF13-GH10 gene in the future.
基金Northern Jiangsu Science and Technology Special Project(XZ-SZ201921)the Technology Program of Gaoyou City(GY201914)。
文摘The Aspergillus niger XynB gene and core promoter region of porcine RELMβgene were cloned into pcDNA3.1(-),and an intestine-specific expression vector pcDNA3.1-RELMβ-XynB-Myc-GFP carrying green fluorescence and Myc double tags was constructed.The vector was transfected into human colon cancer cells(HT29)and human liver cancer cells(Bel7402)using liposomes.Fluorescence microscopy revealed that the vector could specifically express green fluorescent protein(GFP)in HT29 cells.RT-PCR and Western Blot were performed on the HT29 cells transfected with the expression vector,and the results showed that the XynB gene was normally transcribed in HT29 cells,and the target protein expression was detected in the cells.
基金This work was supported by the State"863"High-Tech Programs(Grant No.2001AA214041).
文摘Extracellular xylanase XYNB from Streptomy-ces olivaceoviridis A1 has been purified and characterized. The optimal pH value and temperature of XYNB for its ac-tivity are 5.2 and 60℃, respectively. The specific activity of XYNB is as high as 2869.78 U/mg. Metal cations, EDTA and SDS have no effects on enzyme activity of XYNB. The gene xynB coding mature protein of XYNB has been cloned by PCR. The forward oligonucleotide primer used in the PCR reaction was synthesized based on the N-terminal amino acid sequence of XYNB mature protein, and the reverse oligonu-cleotide primers are random oligonucleotide. The cloned gene xynB is 576 bp long and its G+C content is 64.3%. The xynB encodes 191 amino acid residues, and the putative mo-lecular weight of XYNB is 20.839 kD. The xynB has been expressed in E. coli, and the expressed xylanase has normal bioactivity.
