牛流行熱是一種藉由節肢動物為媒介所引起的病毒性疾病，可引發乳牛、黃牛及水牛的急性發熱感染症，造成重大經濟損失。台灣曾於西元1967、1983、1989、1996、1999、2001及2002年發生過七次牛流行熱大流行，而且疫情爆發間隔漸漸縮短、每次疫情之發病期也逐漸延長、臨床症狀也較過去嚴重，因此加強牛流行熱預防接種是極為重要。台灣雖然於1984年開始使用由呂榮修等人製成的死毒疫苗迄今，但疫情依然不斷發生，檢討原因可能與疫苗接種率低、接種時機不對或病毒株變異有關。牛流行熱之病原為牛流行熱病毒(bovine ephemeral fever virus, BEFV)，在分類上屬於子彈型病毒科(rhabdoviridae)的暫時熱病毒屬(ephemerovirus)。BEFV病毒具有外套膜(envelope)，基因組為負性單股RNA，有6個基因：N、P、M、G、L及GNS，其中前5個是存在於病毒顆粒中，GNS醣蛋白則只在受病毒感染的細胞中偵測到，不存於病毒顆粒中，其功能未知，有文獻認為它是G基因之重複(gene duplication)(Wang and Walker, 1993)。G為病毒外套膜醣蛋白，具型別專一性及中和性抗原簇，並可引發中和性免疫反應。

　　本實驗室曾對數種不同年代BEFV台灣分離株G基因進行變異分析，本研究即擬進一步分析GNS基因之變異及其可能扮演之角色，結果顯示不同年代分離之病毒GNS基因序列亦會與G基因類似具小幅度改變。1984、1996、1999及2001年分離株GNS基因之間核酸及胺基酸序列相似性分別為96.6％〜99.3％、97.8％〜99.3％，G基因則為97.6％〜99.7％、97.9％〜99.5％，皆呈微小差異。而GNS與G之序列比較發現各分離株核酸及胺基酸序列相似度僅為25.4％〜27.0％、13.5％〜14.0％，相似性差異很大。接著取2001年分離株於BHK-21細胞培養，檢定不同時間點之GNS與G基因的表現情形，結果發現G基因表現訊息比GNS基因早，GNS之表現似乎不是G基因表現之所必須。綜合上述二者結果顯示GNS之功能似與G無關，也不太可能是G基因重複而來。但GNS確實亦隨年代在變異，值得進一步探討其功能。

　Bovine ephemeral fever is an arthropod-borne viral disease which causes an acute febrile infection in dairy cattle, yellow cattle and water buffalo. The disease always generates significant economic impairment. There were 7 epizootics occurring in 1967, 1983, 1989, 1996, 1999, 2001, and 2002 in Taiwan. The interval of outbreak is becoming shortened, the duration of infection is gradually prolonged, and the clinical signs are more severe in recent epidemics, it’s very important to strengthen vaccination. These epizootics still occurred despite vaccination strategy with inactivated virus since 1984. The reasons may be the vaccination policy was not efficiently fulfilled or a variant virus appeared. Bovine ephemeral fever virus (BEFV) has been classified as the genus Ephemerovirus in Rhabdoviridae family. The virions are bullet-shaped, enveloped, and contain a negative single-stranded RNA genome with six genes: N, P, M, G, L and GNS. The first five genes are present inside virion. The GNS protein is only found inside BEFV-infected cells with function unknown. The G protein is an envelope-associated glycoprotein containing type-specific and neutralizing epitopes. A gene duplicate of G gene has been proposed (Wang and Walker, 1993).

　The G gene variation in Taiwan isolates were analyzed in our previous study. This study was aimed to analyze the GNS gene variation and it’s probable function. Our results indicated that the GNS genes of Taiwan isolates (isolated in 1984, 1996, 1999, 2001) also showed small variation as the G genes did. Similarities for the nucleotide and amino acid sequences of GNS genes were ranged from 96.6％-99.3％ and 97.8％-99.3％, respectively. Those for G genes were 97.6％-99.7％ and 97.9％-99.5％, respectively. Both GNS and G genes of different isolates showed small variations according to epidemic times. Similarities for the nucleotide and amino acid sequences between GNS and G gene were, however, ranged from 25.4％-27.0％ and 13.5％-14.0％. Big difference was found. Furthermore, we investigated the GNS and G gene expression in BEFV-infected BHK-21 cells. We found that the G gene expression is always earlier than GNS gene indicating that the GNS gene pre-expression is not necessary for G gene expression. In summary, it seems that the GNS gene is not a gene duplicate of G gene. Further study of GNS gene function is needed.

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