Trobridge G D, Leong J A
Department of Microbiology, Oregon State University, Corvallis 97331-3804, USA.
J Interferon Cytokine Res. 1995 Aug;15(8):691-702. doi: 10.1089/jir.1995.15.691.
A full-length cDNA clone of a rainbow trout (Oncorhynchus mykiss) Mx gene was obtained using RACE (rapid amplification of cDNA ends) polymerase chain reaction (PCR) amplification of RNA extracted from poly (I).(C)-induced rainbow trout gonad cells (RTG-2). Mx was previously identified in rainbow trout by Staeheli et al. by hybridization with a partial perch genomic Mx probe to induced rainbow trout mRNA. The 2.5 kb rainbow trout cDNA clone contains an open reading frame of 1863 nt (nucleotides) encoding a 621 amino acid protein. The deduced rainbow trout Mx protein is 70.6 kD and contains the characteristic tripartite GTP binding motif common to all Mx protein. Southern blot analysis with the rainbow trout Mx probe demonstrated the presence of Mx homologous genes in four other salmonid fish species, including chinook, coho, and kokanee salmon and brook trout. Poly (I).(C) treatment of both RTG-2 and chinook salmon cells (CHSE-214) induced two transcripts whose appearance was observed first at 24 h and as long as 72 h after treatment. Infection of rainbow trout with the salmonid rhabdovirus, IHNV (infectious hematopoietic necrosis virus), also induced the synthesis of Mx mRNA. A comparison of the rainbow trout Mx protein with other reported Mx proteins indicates that the piscine Mx is highly homologous to the mammalian Mx proteins.
利用从聚(I)·(C)诱导的虹鳟性腺细胞(RTG - 2)中提取的RNA,通过cDNA末端快速扩增(RACE)聚合酶链反应(PCR)扩增,获得了虹鳟(Oncorhynchus mykiss)Mx基因的全长cDNA克隆。此前,Staeheli等人通过用部分鲈鱼基因组Mx探针与诱导的虹鳟mRNA杂交,在虹鳟中鉴定出了Mx。这个2.5 kb的虹鳟cDNA克隆包含一个1863个核苷酸(nt)的开放阅读框,编码一个621个氨基酸的蛋白质。推导的虹鳟Mx蛋白为70.6 kD,含有所有Mx蛋白共有的特征性三联体GTP结合基序。用虹鳟Mx探针进行的Southern印迹分析表明,在其他四种鲑科鱼类中存在Mx同源基因,包括奇努克鲑、银鲑、红大马哈鱼和溪鳟。对RTG - 2细胞和奇努克鲑细胞(CHSE - 214)进行聚(I)·(C)处理,诱导产生了两种转录本,在处理后24小时首次观察到它们的出现,并且在处理后长达72小时都能观察到。用鲑科弹状病毒传染性造血坏死病毒(IHNV)感染虹鳟,也诱导了Mx mRNA的合成。将虹鳟Mx蛋白与其他已报道的Mx蛋白进行比较表明,鱼类Mx与哺乳动物Mx蛋白高度同源。