Analysis and Expression of Tumor Necrosis Factor Receptor Homolog from LCDV-C

HUNG You-hua; SUN Wei; ZHANG Qi-ya*

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December 2005

HUNG You-hua, SUN Wei and ZHANG Qi-ya*. Analysis and Expression of Tumor Necrosis Factor Receptor Homolog from LCDV-C[J]. Virologica Sinica, 2005, 20(6): 652-655.

Citation: HUNG You-hua, SUN Wei, ZHANG Qi-ya*. Analysis and Expression of Tumor Necrosis Factor Receptor Homolog from LCDV-C .VIROLOGICA SINICA, 2005, 20(6) : 652-655.

淋巴囊肿病毒中国株TNFR类似物的原核表达与结构分析

1.
1.中国科学院水生生物研究所淡水生态与生物技术国家重点实验室湖北武汉 2.中国科学院研究生院北京

摘要

肿瘤坏死因子受体(TNFR)是细胞因子受体家族中的一员,在大DNA病毒的免疫逃避中起着重要的作用。淋巴囊肿病毒中国株(LCDV-C)是一种大DNA病毒,属于虹彩病毒科。参照已知虹彩病毒TNFR基因设计引物: P1,5′GGATCCAAAACTATGATTAAAATAAAGA 3′;P2:5′ATTACTCGAGAATGTTAAAAATTAAGCTT 3′。以LCDV-C基因组DNA为模板,PCR扩增得到一个834bp的DNA片段,并对该片段进行测序。构建原核表达重组质粒后,在大肠杆菌DE3中诱导表达,其产物经SDS-PAGE电泳后,显示为45kDa的融合蛋白带。对测序结果进行计算机辅助分析的结果显示,LCDV-C TNFR类似物是一个含278个氨基酸的多肽,具有典型的半胱氨酸富集区功能结构域,与宿主牙鲆TNFRII氨基酸同源性为34％。
- 淋巴囊肿病毒
- , 虹彩病毒
- , 肿瘤坏死因子受体类似物
- , 序列分析
- , 原核表达

Analysis and Expression of Tumor Necrosis Factor Receptor Homolog from LCDV-C

Abstract

Tumor necrosis factor receptor (TNFR) plays an important role in the evasion of immune response by large DNA viruses. TNFR is a homologue of cellular receptors. Lymphocystis disease virus, an iriduvirus, isolated in China (LCDV-C) is a large DNA virus. Primers were designed based on conserved TNFR nucleotide sequences in iridoviruses and used to amplify the homologue in LCDV-C. A DNA fragment of 834 bp was generated and sequenced. The recombi-nant prokaryotic expression plasmid containing this fragment was constructed and expressed in E. coli. DE3. An expected 45kDa fusion protein was isolated by SDS-PAGE. Computer analysis indicated that LCDV-C TNFR homologue encodes a 278aa putative protein, which contains a typical cysteine-rich domain. It shares 34% identity with flounder TNFRII.
- Lymphocystis disease virus
- , Iridovirus
- , TNFR homolog
- , Sequence analysis
- , Prokaryotic expression

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

Analysis and Expression of Tumor Necrosis Factor Receptor Homolog from LCDV-C

Abstract: Tumor necrosis factor receptor (TNFR) plays an important role in the evasion of immune response by large DNA viruses. TNFR is a homologue of cellular receptors. Lymphocystis disease virus, an iriduvirus, isolated in China (LCDV-C) is a large DNA virus. Primers were designed based on conserved TNFR nucleotide sequences in iridoviruses and used to amplify the homologue in LCDV-C. A DNA fragment of 834 bp was generated and sequenced. The recombi-nant prokaryotic expression plasmid containing this fragment was constructed and expressed in E. coli. DE3. An expected 45kDa fusion protein was isolated by SDS-PAGE. Computer analysis indicated that LCDV-C TNFR homologue encodes a 278aa putative protein, which contains a typical cysteine-rich domain. It shares 34% identity with flounder TNFRII.