Cloning and Sequencing of the Full-length cDNA of Mx Protein from Mandarinfish

WU Hai-feng; BAI Jun-jie; LAO Hai-hua; YE Xing; JIAN Qing; LUO Jian-ren

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June 2004

WU Hai-feng, BAI Jun-jie, LAO Hai-hua, YE Xing, JIAN Qing and LUO Jian-ren. Cloning and Sequencing of the Full-length cDNA of Mx Protein from Mandarinfish[J]. Virologica Sinica, 2004, 19(3): 271-275.

Citation: WU Hai-feng, BAI Jun-jie, LAO Hai-hua, YE Xing, JIAN Qing, LUO Jian-ren. Cloning and Sequencing of the Full-length cDNA of Mx Protein from Mandarinfish .VIROLOGICA SINICA, 2004, 19(3) : 271-275.

鳜鱼Mx蛋白全长cDNA的克隆和序列分析*

吴海峰 ^1,2 ,
白俊杰 ¹ ,
劳海华 ¹ ,
罗建仁 ¹ ,

1.
1. 中国水产科学研究院珠江水产研究所,中国水产科学研究院热带亚热带鱼类选育与养殖重点开放实验室，广东广州 510380
2.
上海水产大学，上海 200090

摘要

Mx蛋白是一类由I型干扰素诱导表达的抗病毒蛋白。本研究以感染了鳜传染性脾肾坏死病毒（Infectious spleen and kidney necrosis virus,ISKNV）的鳜鱼为材料，提取肝脏总RNA，通过逆转录-聚合酶链式反应（RT-PCR）扩增出Mx蛋白基因的核心片段序列，再应用3’和5’快速扩增cDNA末端（RACE）方法PCR扩增Mx蛋白cDNA末端，最终获得鳜鱼Mx蛋白cDNA序列（GenBank登陆号：AY392097）。序列分析表明：鳜鱼Mx蛋白cDNA含有2391bp，其中编码区长1881bp，编码627个氨基酸残基，推测蛋白质分子量大小为7.15kDa。鳜鱼Mx蛋白具有脊椎动物Mx蛋白共有的结构特征：一个三联体GTP结合区域（GXXXSGKS/T、DXXG、T/NKXD）；一个发动蛋白家族的典型结构特征序列（LPRGS/KGIVTR）；以及C端高度保守的Leu拉链结构域。鳜鱼Mx蛋白全基因的获得为下一步研究鱼类Mx蛋白的抗病毒活性、作用机制，以及干扰素的检测奠定了基础。
- 鳜鱼
- , Mx蛋白
- , RACE
- , 全长cDNA
- , 序列分析

Cloning and Sequencing of the Full-length cDNA of Mx Protein from Mandarinfish

WU Hai-feng ^1,2 ,
BAI Jun-jie ¹ ,
LAO Hai-hua ¹ ,
YE Xing ¹ ,
JIAN Qing ¹ ,
LUO Jian-ren ^1*,,

1.
1、Pearl River Fisheries Research Institute, Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation, Chinese Academy of Fishery Sciences, Guangzhou 510380
2.
Shanghai Fisheries University, Shanghai 200090

Corresponding author: LUO Jian-ren,

Abstract

Mx protein is one of the classes of antiviral proteins induced by type I interferons (IFN). A full-length cDNA of mandarinfish (Siniperca chuatsi) Mx gene was obtained using reverse transcription polymerase chain reaction (RT-PCR) and RACE (rapid amplification of cDNA ends) polymerase chain reaction (PCR) amplification of total RNA extracted from liver of a mandarinfish which was infected with Infectious spleen and kidney necrosis virus (ISKNV) (GenBank accession No.AY392097). The result of sequence analysis indicates that the 2.4kb mandarinfish Mx cDNA contains an open reading frame (ORF) of 1881nt encoding a 627 amino acid protein whose deduced protein molecular weight is 71.5kDa. The alignment of the deduced mandarinfish Mx protein amino acid sequence with that of other species shows that the Mx protein of mandarinfish contains a triparitite guanosine-5’-triphosphate (GTP)-binding motif (GXXXSGKS/T, DXXG, T/NKXD), a signature of the dynamin family (LPRGS/ KGIVTR), and a highly conserved leucine zipper motif in the C-terminal region of the protein, which are conserved in all IFN-induced Mx proteins of vertebrates.
- Siniperca chuatsi
- , Mx protein
- , RACE
- , Full-length cDNA
- , Sequence analysis

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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

Cloning and Sequencing of the Full-length cDNA of Mx Protein from Mandarinfish

Corresponding author: LUO Jian-ren,

1. 1、Pearl River Fisheries Research Institute, Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation, Chinese Academy of Fishery Sciences, Guangzhou 510380
2. Shanghai Fisheries University, Shanghai 200090

Abstract: Mx protein is one of the classes of antiviral proteins induced by type I interferons (IFN). A full-length cDNA of mandarinfish (Siniperca chuatsi) Mx gene was obtained using reverse transcription polymerase chain reaction (RT-PCR) and RACE (rapid amplification of cDNA ends) polymerase chain reaction (PCR) amplification of total RNA extracted from liver of a mandarinfish which was infected with Infectious spleen and kidney necrosis virus (ISKNV) (GenBank accession No.AY392097). The result of sequence analysis indicates that the 2.4kb mandarinfish Mx cDNA contains an open reading frame (ORF) of 1881nt encoding a 627 amino acid protein whose deduced protein molecular weight is 71.5kDa. The alignment of the deduced mandarinfish Mx protein amino acid sequence with that of other species shows that the Mx protein of mandarinfish contains a triparitite guanosine-5’-triphosphate (GTP)-binding motif (GXXXSGKS/T, DXXG, T/NKXD), a signature of the dynamin family (LPRGS/ KGIVTR), and a highly conserved leucine zipper motif in the C-terminal region of the protein, which are conserved in all IFN-induced Mx proteins of vertebrates.