Endothelin(ET)is the most potent mammalian vasoconstrictor identified to data.As a pathogenic factor,ET is involved in the genesis of many diseases.In this study,a pair of primers was designed and synthesized accordin...Endothelin(ET)is the most potent mammalian vasoconstrictor identified to data.As a pathogenic factor,ET is involved in the genesis of many diseases.In this study,a pair of primers was designed and synthesized according to the human ETB receptor gene (hETBR)sequence.A 394bp of DNA fragment was amplified by polymerase chain reaction(PCR) and labeled with α 32 P CTP using Random Primer Labeling method.With this probe,rabbit lung cDNA library was screened by in situ hybridization and 11 positive clones were identified.Sequencing result showed that a complete reading frame of rabbit ETB receptor(rETBR)cDNA could be produced from three positive clones of eleven.By a series of subcloning,a recombinant plasmid including the 1326 bp of rETBR coding sequences,named pBlu Script rETBR,was constructed.The deduced amino acid sequence indicated that the rETBR is 441 residues in length,with an expected molecular mass of approximately 49.44 kD.N terminal 18 residues is the potential signal peptide (Score=11.11)and therefore the molecular mass of mature rETBR is 47.65 kD with 423 amino acid residues.Analysis of the rETBR hydropathy profile indicates the presence of seven hydrophobic regions,putative transmembrane domains.Potential N glycosylation sites are the 60th and the 118 th.The structure exhibits a significant sequence and topographical similarity with G protein coupled receptors.展开更多
A gene library of the infectious spleen and kidney necrosis virus(ISKNV) from mandarinfish,Siniperca chuatsi(Basilewsky) genome was established ISKNV DNA was cleaved with EcoRI,BamHI,HindⅢ,KpnI and PstI and the resul...A gene library of the infectious spleen and kidney necrosis virus(ISKNV) from mandarinfish,Siniperca chuatsi(Basilewsky) genome was established ISKNV DNA was cleaved with EcoRI,BamHI,HindⅢ,KpnI and PstI and the resulting fragments were inserted into the corresponding sites of the pBluescript Ⅱ KS plasmid vector using T4 DNA ligase Bacterial colonies harboring recombinant plasmids were selected All cloned fragments were individually identified by digestion of the recombinant plasmid DNA with different restriction enzymes and screened by hybridization of recombinant plasmid DNA to viral DNA Using these recombinant plasmids,the physical maps of the genome were constructed for BamHI,HindⅢ and KpnI restriction展开更多
文摘Endothelin(ET)is the most potent mammalian vasoconstrictor identified to data.As a pathogenic factor,ET is involved in the genesis of many diseases.In this study,a pair of primers was designed and synthesized according to the human ETB receptor gene (hETBR)sequence.A 394bp of DNA fragment was amplified by polymerase chain reaction(PCR) and labeled with α 32 P CTP using Random Primer Labeling method.With this probe,rabbit lung cDNA library was screened by in situ hybridization and 11 positive clones were identified.Sequencing result showed that a complete reading frame of rabbit ETB receptor(rETBR)cDNA could be produced from three positive clones of eleven.By a series of subcloning,a recombinant plasmid including the 1326 bp of rETBR coding sequences,named pBlu Script rETBR,was constructed.The deduced amino acid sequence indicated that the rETBR is 441 residues in length,with an expected molecular mass of approximately 49.44 kD.N terminal 18 residues is the potential signal peptide (Score=11.11)and therefore the molecular mass of mature rETBR is 47.65 kD with 423 amino acid residues.Analysis of the rETBR hydropathy profile indicates the presence of seven hydrophobic regions,putative transmembrane domains.Potential N glycosylation sites are the 60th and the 118 th.The structure exhibits a significant sequence and topographical similarity with G protein coupled receptors.
文摘A gene library of the infectious spleen and kidney necrosis virus(ISKNV) from mandarinfish,Siniperca chuatsi(Basilewsky) genome was established ISKNV DNA was cleaved with EcoRI,BamHI,HindⅢ,KpnI and PstI and the resulting fragments were inserted into the corresponding sites of the pBluescript Ⅱ KS plasmid vector using T4 DNA ligase Bacterial colonies harboring recombinant plasmids were selected All cloned fragments were individually identified by digestion of the recombinant plasmid DNA with different restriction enzymes and screened by hybridization of recombinant plasmid DNA to viral DNA Using these recombinant plasmids,the physical maps of the genome were constructed for BamHI,HindⅢ and KpnI restriction
