狂犬病病毒CTN-1株在鸡胚细胞驯化过程中G基因和滴度分析

发布时间：2018-10-26 18:18

【摘要】：目的研究狂犬病病毒CTN-1株在鸡胚细胞驯化过程中的特性。方法利用RT-PCR反应扩增CTN-1株19个代次的G蛋白基因,获得cDNA进行序列测定;并以免疫荧光法测定各代次毒株在细胞中的滴度。结果序列测定显示:CTN-1株在鸡胚细胞驯化过程中G蛋白基因序列发生不同程度变异且逐步积累,从第30代开始,G蛋白基因序列趋于稳定。与CTN-1株相比,CTN-1鸡胚细胞驯化株在G蛋白的第147、333、389、421和485位氨基酸发生了突变。与此相对应,CTN-1株鸡胚细胞驯化株滴度逐渐趋于稳定,并于第33-58代滴度维持在10~101g细胞荧光转化灶单位/mL左右。G蛋白同源性分析表明,CTN-1株鸡胚细胞驯化株与我国近期不同区域分离得到的街毒株具有较高同源性,其G蛋白氨基酸同源性为89.3%~99.0%。结论 CTN-1株在鸡胚细胞驯化过程中G蛋白基因发生稳定变异,滴度在传代后33-58代达到较高水平,稳定性较好,适于我国狂犬病防治的实际应用。
[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.
【作者单位】：深圳市卫光生物制品股份有限公司;
【分类号】：R512.99

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