Purification of Glutathione S-transferase Fusion Proteins of SARS Coronavirus Spike Protein  ragment 2 and Preparation of Anti-GST-S2 Polyclonal Antibody

WANG Da-yan; WANG Jian-wei*; WEi Qiang; QU Jian-guo; HONG Tao

PDF
Cite
Share

facebook

twitter

google

linkedin

June 2005

WANG Da-yan, WANG Jian-wei*, WEi Qiang, QU Jian-guo and HONG Tao. Purification of Glutathione S-transferase Fusion Proteins of SARS Coronavirus Spike Protein ragment 2 and Preparation of Anti-GST-S2 Polyclonal Antibody[J]. Virologica Sinica, 2005, 20(3): 247-252.

Citation: WANG Da-yan, WANG Jian-wei*, WEi Qiang, QU Jian-guo, HONG Tao. Purification of Glutathione S-transferase Fusion Proteins of SARS Coronavirus Spike Protein ragment 2 and Preparation of Anti-GST-S2 Polyclonal Antibody .VIROLOGICA SINICA, 2005, 20(3) : 247-252.

我国人轮状病毒不同基因型NSP4的致病性研究

1.
中国疾病预防控制中心病毒病预防控制所 100052北京

摘要

对我国轮状病毒流行株NSP4基因变异特点的分析表明,NSP4基因主要可分为Wa组和Kun组,在Wa组内可形成三个亚组,形成了4种NSP4基因型。为了进一步阐明人轮状病毒流行株NSP4基因变异与其致病性变化是否存在联系,我们首先利用杆状病毒载体对NSP4蛋白进行表达,获得了对应4种不同NSP4基因型的重组杆状病毒rvBac97B6,rvBac97S34,rvBac97S36和rvBac97SZ8。用这些病毒感染Sf9细胞后,检测细胞内Ca2+浓度的变化,发现与野生型杆状病毒感染细胞相比,重组病毒感染细胞内的Ca2+浓度显著升高,但各个重组病毒之间无显著性差异。在此基础上,我们进一步在E.coli中分别表达纯化了代表Wa和Kun基因分组的97S34和97SZ8流行株的NSP4。分别用纯化的重组NSP4蛋白攻击乳鼠后,发现不同基因型的NSP4蛋白的致腹泻活性没有明显差异,这种作用可被NSP4抗体拮抗,但这种拮抗作用存在基因型特异性。上述结果表明人轮状病毒流行株NSP4氨基酸序列间的变异并没有使其钙调节及致腹泻能力产生改变,在致腹泻作用中发挥关键作用(或决定性作用)的氨基酸位点在不同NSP4基因型间可能是相对保守的。针对NSP4抗体的有效性也为新型轮状病毒疫苗和药物研究提供了线索。
- 轮状病毒
- , NSP4
- , 变异
- , Ca2+
- , 腹泻

Purification of Glutathione S-transferase Fusion Proteins of SARS Coronavirus Spike Protein ragment 2 and Preparation of Anti-GST-S2 Polyclonal Antibody

Abstract

Our previous studies showed that the NSP4 gene of human rotavirus epidemic strains in China can be divided into two groups:Wa and Kun, and the Wa group contains three subgroups. To investigate whether there were differences in virulence among the different rotavirus NSP4 genotypes, NSP4 genes from different genetic types were expressed in recombinant baculoviruses using Bac-to-Bac expression system and four recombinant baculoviruses were obtained. Compared with the mock infected cells or the wild type baculovirus AcNPV infected cells, the intracellular Ca 2+concentration increased 5.24, 5.43, 5.34 and 5.45 folds at 72h post infection, respectively. However, there was no significant difference among the increasing of intracellular calcium level induced by the four recombinant baculoviruses. On the basis of the above results, the recombinant NSP4 proteins from Wa or Kun groups were expressed in E.coli and purified ,and the diarrhea induction activity was tested in sucking mice model. The results showed that dia
- Rotavirus
- , NSP4
- , Variation
- , Ca 2+
- , Diarrhea

References
Proportional views

PDF

Article Metrics

Article views(4279) PDF downloads(1052) Cited by(0)

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Purification of Glutathione S-transferase Fusion Proteins of SARS Coronavirus Spike Protein ragment 2 and Preparation of Anti-GST-S2 Polyclonal Antibody

Abstract: Our previous studies showed that the NSP4 gene of human rotavirus epidemic strains in China can be divided into two groups:Wa and Kun, and the Wa group contains three subgroups. To investigate whether there were differences in virulence among the different rotavirus NSP4 genotypes, NSP4 genes from different genetic types were expressed in recombinant baculoviruses using Bac-to-Bac expression system and four recombinant baculoviruses were obtained. Compared with the mock infected cells or the wild type baculovirus AcNPV infected cells, the intracellular Ca 2+concentration increased 5.24, 5.43, 5.34 and 5.45 folds at 72h post infection, respectively. However, there was no significant difference among the increasing of intracellular calcium level induced by the four recombinant baculoviruses. On the basis of the above results, the recombinant NSP4 proteins from Wa or Kun groups were expressed in E.coli and purified ,and the diarrhea induction activity was tested in sucking mice model. The results showed that dia