【摘要】：目的研究狂犬病病毒CTN-1株在雞胚細(xì)胞馴化過程中的特性。方法利用RT-PCR反應(yīng)擴(kuò)增CTN-1株19個(gè)代次的G蛋白基因,獲得cDNA進(jìn)行序列測(cè)定;并以免疫熒光法測(cè)定各代次毒株在細(xì)胞中的滴度。結(jié)果序列測(cè)定顯示:CTN-1株在雞胚細(xì)胞馴化過程中G蛋白基因序列發(fā)生不同程度變異且逐步積累,從第30代開始,G蛋白基因序列趨于穩(wěn)定。與CTN-1株相比,CTN-1雞胚細(xì)胞馴化株在G蛋白的第147、333、389、421和485位氨基酸發(fā)生了突變。與此相對(duì)應(yīng),CTN-1株雞胚細(xì)胞馴化株滴度逐漸趨于穩(wěn)定,并于第33-58代滴度維持在10~101g細(xì)胞熒光轉(zhuǎn)化灶單位/mL左右。G蛋白同源性分析表明,CTN-1株雞胚細(xì)胞馴化株與我國近期不同區(qū)域分離得到的街毒株具有較高同源性,其G蛋白氨基酸同源性為89.3%~99.0%。結(jié)論 CTN-1株在雞胚細(xì)胞馴化過程中G蛋白基因發(fā)生穩(wěn)定變異,滴度在傳代后33-58代達(dá)到較高水平,穩(wěn)定性較好,適于我國狂犬病防治的實(shí)際應(yīng)用。
[Abstract]:Objective to study the characteristics of rabies virus CTN-1 strain during the domestication of chicken embryo cells. Methods the G protein genes of 19 generations of CTN-1 strain were amplified by RT-PCR reaction and sequenced by cDNA, and the titers of each generation were determined by immunofluorescence method. Results the sequence analysis showed that the G protein gene sequence of CTN-1 strain changed and accumulated gradually in the process of chicken embryo cell acclimation. From the 30th generation, the G protein gene sequence tended to be stable. Compared with CTN-1 strain, CTN-1 chicken embryo cell domesticated strain was mutated at the amino acid 147333389421 and 485 of G protein. Correspondingly, the titer of the domesticated strain of chicken embryo cells of CTN-1 strain tended to be stable, and the titer of 33-58 generation cells was maintained at about 10 ~ 101g / mL. The homology analysis of G protein showed that, The domesticated strain of chicken embryo cells of CTN-1 strain had high homology with the street virus strain isolated from different regions of China recently, and the amino acid homology of G protein was 89.33% 99.0%. Conclusion the G protein gene of CTN-1 strain has stable variation during the process of chicken embryo cell acclimation, and the titer reaches a higher level in the 33-58 passages after passage, which is suitable for the practical application of rabies prevention and treatment in China.
【作者單位】： 深圳市衛(wèi)光生物制品股份有限公司;
【分類號(hào)】：R512.99

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