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題名 | 臺灣雞傳染性喉頭氣管炎野外株與疫苗株病毒醣蛋白之分析= |
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作者 | 沈瑞鴻; 謝快樂; |
期刊 | 臺灣畜牧獸醫學會會報 |
出版日期 | 19921200 |
卷期 | 60 1992.12[民81.12] |
頁次 | 頁1-13 |
分類號 | 437.717 |
語文 | chi |
關鍵詞 | 疫苗; 株; 氣管炎; 病毒; 野外; 喉頭; 傳染性; 臺灣; 醣蛋白; 雞; |
中文摘要 | 自本省田間分離雞傳染性喉頭氣管炎(ILT)病毒株和本省普遍使用之3株疫苗 株,經篩選,分別於雞胚腎臟細胞增殖、純化,以十二烷硫酸鈉(Sodium dodecyl sulfate) -聚丙烯醯膠體(Polyacrylamide gel)電泳分析(SDS-PAGE),顯示各病毒株結構蛋白質的分 子量,分別為250kDa、16OkDa、ll5kDa、9OkDa、6OkDa、34kDa、和26KDa,比較在膠體 上的移動圖譜,顯示株間,並無明顯差異。同時亦分別就野外分離株及疫苗株,純化之完整 病毒顆粒,經Triton X-l00處理後,通過Con A親和性管柱,以純化病毒醣蛋白,以 SDS-PAGE分析,並經西方轉印後與抗ILT病毒之多價抗體作用,顯示純化所得為分子量 205kDa、115kDa、9OkDa、6OkDa之病毒醣蛋白。 另以純化之ILT病毒為抗原,研製得3株抗此病毒之單株抗體融合瘤,以西方轉印法證 明他們均可與分子量為6OkDa之病毒蛋白結合。 |
英文摘要 | Attemps of this study were made to compare viral glycoprotein among field ILTV and vaccine ILT strains; to purify the viral glycoprotein by suing the Con-A sepharose chromatography and to produce their antibodies in mice; to establish hybridoma which produce antibodies against the viral glycoprotein. Field isolate and vaccine ILTV were propagated in CEK cell cultures, purified by means of sucrose gradient centrifugation, then subjected to SDS-PAG electrophoresis analysis and Coomassie blue stain. Protein bands with molecular weight 205, 160, 115, 90, 60, 34 and 26 Kd were found in all the ILTV strains analysed. Cloned field ILTV and vaccine strains were propagated and purified as mentioned above, then treated with Triton X-100, respectively. The suspension were then ultracentrifugated. The viral glycoprotein contained supernatants were further passed through Con-A sepharose chromatography. The eluted pure viral proteins were subjected to SDS-PAGE, then analyzed by means of Coomassie blue, or by means of western blotting then reacted with ILTV hyperimmune serum and detected with peroxidase conjugated anti-α-globulin. The purified viral glycoprotein were noted at molecular weights 205, 115, 90 and 60 Kd bands. Spleen cells of pure ILTV vaccinated BALB/c mice were harvested and fused with NS-1 myeloma cells. Antibodies from these fused hybridoma were detected by means of ELISA and indirect fluorescent antibody stain by using pure ILTV or ILTV cultured CEK, respectively. Twenty-two hybridoma were positive in ILTV antibody production. However, only three out of the 22 hybridoma could continuously grow in HAT maintenance medium and continuously or in vivo mice ascites were subjected to SDS-PAGE and western blotting analysis. The molecular weight of the antibodies produced by all three clones of hybridoma was all the band 60 Kd |
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