前期研究发现,信号淋巴细胞活化分子(signaling lymphocyte activation molecule,SLAM or CD150)和脊髓灰质炎病毒受体相关蛋白4(poliovirus receptor-related protein 4,PVRL4 or Nectin4)是麻疹病毒属副黏病毒的主要细胞受体,也参与...前期研究发现,信号淋巴细胞活化分子(signaling lymphocyte activation molecule,SLAM or CD150)和脊髓灰质炎病毒受体相关蛋白4(poliovirus receptor-related protein 4,PVRL4 or Nectin4)是麻疹病毒属副黏病毒的主要细胞受体,也参与禽副黏病毒NDV感染的感染过程[1]。为了验证鸡源Nectin4(chNectin4)和鸡源SLAM(chSLAM)在NDV感染中的作用,本研究构建了分别稳定表达chNectin4和chSLAM的BHK21细胞系。感染实验结果显示,稳定表达chNectin4和chSLAM细胞系在接种NDV La Sota株后12 h和24 h NP mRNA水平显著高于亲本细胞,表明chNectin4和chSLAM有助于促进NDV在BHK21细胞系中的增殖,其中chNectin4的促NDV增殖作用更加明显。本研究为进一步研究鸡Nectin4和SLAM在NDV感染中的作用以及NDV疫苗的研究奠定了基础。展开更多
Lumbrukinase gene from earthworm ( L.bimastus ) was obtained by RT\|PCR. The product, PI 239 , was sequenced and analyzed by biology programs and database. The gene including signal peptide coding sequence was cloned ...Lumbrukinase gene from earthworm ( L.bimastus ) was obtained by RT\|PCR. The product, PI 239 , was sequenced and analyzed by biology programs and database. The gene including signal peptide coding sequence was cloned into an eukaryotic vector and the clones were obtained by transferring into BHK cells. The gene was fused with EGFP gene at C\|terminal to be detected conveniently by its fluorescence. The lumbrukinase gene PI 239 has 852 nucleotides that code for 239 amino acid residues as mature peptide chain. The N terminal of PI 239 shares certain homology with those known lumbrukinase. The enzyme contains relative more acidic amino acid residues, and has homology to serine protease. It belongs to the acidic protein, serine protease. Conformation prediction indicates that its secondary structure mainly consists of β sheet. It has two super secondary structure motifs with the active sites Asp188 and Ser189 in between. The DNA and mRNA of the whole gene could be detected in BHK clones, but no recombinant protein detected. Under cofocal microscope, the cells transferred with fused gene showed fluorescence indicating that the fused protein was expressed. In addition, the cells containing the fused gene died soon while most of the control cells were still alive. It seemed that the protein could be expressed in BHK cells as a cytotoxin, though at a low level.展开更多
文摘前期研究发现,信号淋巴细胞活化分子(signaling lymphocyte activation molecule,SLAM or CD150)和脊髓灰质炎病毒受体相关蛋白4(poliovirus receptor-related protein 4,PVRL4 or Nectin4)是麻疹病毒属副黏病毒的主要细胞受体,也参与禽副黏病毒NDV感染的感染过程[1]。为了验证鸡源Nectin4(chNectin4)和鸡源SLAM(chSLAM)在NDV感染中的作用,本研究构建了分别稳定表达chNectin4和chSLAM的BHK21细胞系。感染实验结果显示,稳定表达chNectin4和chSLAM细胞系在接种NDV La Sota株后12 h和24 h NP mRNA水平显著高于亲本细胞,表明chNectin4和chSLAM有助于促进NDV在BHK21细胞系中的增殖,其中chNectin4的促NDV增殖作用更加明显。本研究为进一步研究鸡Nectin4和SLAM在NDV感染中的作用以及NDV疫苗的研究奠定了基础。
文摘Lumbrukinase gene from earthworm ( L.bimastus ) was obtained by RT\|PCR. The product, PI 239 , was sequenced and analyzed by biology programs and database. The gene including signal peptide coding sequence was cloned into an eukaryotic vector and the clones were obtained by transferring into BHK cells. The gene was fused with EGFP gene at C\|terminal to be detected conveniently by its fluorescence. The lumbrukinase gene PI 239 has 852 nucleotides that code for 239 amino acid residues as mature peptide chain. The N terminal of PI 239 shares certain homology with those known lumbrukinase. The enzyme contains relative more acidic amino acid residues, and has homology to serine protease. It belongs to the acidic protein, serine protease. Conformation prediction indicates that its secondary structure mainly consists of β sheet. It has two super secondary structure motifs with the active sites Asp188 and Ser189 in between. The DNA and mRNA of the whole gene could be detected in BHK clones, but no recombinant protein detected. Under cofocal microscope, the cells transferred with fused gene showed fluorescence indicating that the fused protein was expressed. In addition, the cells containing the fused gene died soon while most of the control cells were still alive. It seemed that the protein could be expressed in BHK cells as a cytotoxin, though at a low level.
