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从大西洋鲑(Salmo salar L.)中克隆双链RNA诱导型Mx基因的分子克隆。

Molecular cloning of double-stranded RNA inducible Mx genes from Atlantic salmon (Salmo salar L.).

作者信息

Robertsen B, Trobridge G, Leong J A

机构信息

Norwegian College of Fishery Science, University of Tromsø, Norway.

出版信息

Dev Comp Immunol. 1997 Sep-Oct;21(5):397-412. doi: 10.1016/s0145-305x(97)00019-0.

Abstract

Mx proteins are induced by type I interferons in mice and humans and inhibit the replication of several negative-stranded RNA viruses. In this work Mx genes in Atlantic salmon were studied using the double stranded RNA, polyinosinic: polycytidylic acid (poly I:C), to induce interferon production. Northern blot analysis showed Mx mRNA in liver, head kidney and gills 2 days after poly I:C injection of fish, but not in untreated fish or fish injected with saline or LPS. Mx transcripts of 2.4 and 1.9 kb were detected in the liver. By screening of a cDNA library, three different full length Mx cDNA clones, ASMx1, ASMx2 and ASMx3, were identified and sequenced. The deduced ASMx proteins all contain 623 amino acids and show the tripartite GTP-binding motif typical of vertebrate Mx proteins. ASMx1 and ASMx2 may represent alleles of the same gene whereas ASMx3 represents a different gene. The deduced ASMx proteins showed 96 to 98% sequence identity with rainbow trout Mx1 and Mx3 and about 88% identity with rainbow trout Mx2 protein.

摘要

Mx蛋白在小鼠和人类中由I型干扰素诱导产生,并抑制多种负链RNA病毒的复制。在本研究中,利用双链RNA聚肌苷酸:聚胞苷酸(poly I:C)诱导干扰素产生,对大西洋鲑的Mx基因进行了研究。Northern印迹分析显示,在给鱼注射poly I:C两天后,肝脏、头肾和鳃中存在Mx mRNA,但未处理的鱼或注射生理盐水或脂多糖的鱼中未检测到。在肝脏中检测到2.4 kb和1.9 kb的Mx转录本。通过筛选cDNA文库,鉴定并测序了三个不同的全长Mx cDNA克隆,即ASMx1、ASMx2和ASMx3。推导的ASMx蛋白均含有623个氨基酸,并显示出脊椎动物Mx蛋白典型的三联体GTP结合基序。ASMx1和ASMx2可能代表同一基因的等位基因,而ASMx3代表不同的基因。推导的ASMx蛋白与虹鳟Mx1和Mx3的序列同一性为96%至98%,与虹鳟Mx2蛋白的同一性约为88%。

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