CAJ | 학술논문

目的测定6代次CTN-1V株病毒糖蛋白基因组序列,并研究其致病力稳定性.方法 RT-PCR方法扩增6代次CTN-1V株病毒糖蛋白基因组序列,分别将产物克隆入pGEM-T载体中,测定并拼接序列,应用DNAStar和Mega4.0软件包中的相应软件对基因组序列进行分析,并与近期我国分离且具有地域代表性的狂犬病病毒街毒株进行基因同源性分析；测定6代次CTN-1V株病毒滴度(LD50/ml)和细胞荧光滴度(FFU/ml),采用FFU/LD50指标来对病毒的毒力进行评价.结果 6代次CTN-1V株病毒序列测定结果表明,仅第1222位的G突变为A,导致408位氨基酸由谷氨酸突变为赖氨酸,而1320位核苷酸由A突变为G,但显示为无义突变；5代次CTN-1V株病毒致病力指标均在1.00～1.07之间；糖蛋白生物信息学分析表明,CTN-1V株病毒与我国近期分离的街毒株均具有较高同源性.结论 CTN-1V株病毒糖蛋白传代稳定,且与我国近期分离的街毒株高度同源,各代次CTN-1V株病毒糖蛋白基因组序列的测定及致病力研究,为完善该毒株的质量控制提供数据支持.
목적 측정6대차CTN-1V주병독당단백기인조서렬,병연구기치병력은정성.방법 RT-PCR방법확증6대차CTN-1V주병독당단백기인조서렬,분별장산물극륭입pGEM-T재체중,측정병병접서렬,응용DNAStar화Mega4.0연건포중적상응연건대기인조서렬진행분석,병여근기아국분리차구유지역대표성적광견병병독가독주진행기인동원성분석；측정6대차CTN-1V주병독적도(LD50/ml)화세포형광적도(FFU/ml),채용FFU/LD50지표래대병독적독력진행평개.결과 6대차CTN-1V주병독서렬측정결과표명,부제1222위적G돌변위A,도치408위안기산유곡안산돌변위뢰안산,이1320위핵감산유A돌변위G,단현시위무의돌변；5대차CTN-1V주병독치병력지표균재1.00～1.07지간；당단백생물신식학분석표명,CTN-1V주병독여아국근기분리적가독주균구유교고동원성.결론 CTN-1V주병독당단백전대은정,차여아국근기분리적가독주고도동원,각대차CTN-1V주병독당단백기인조서렬적측정급치병력연구,위완선해독주적질량공제제공수거지지.
Objective To sequence the genes encoding glycoprotein (G) of rabies virus strain CTN-1V after 6 passages,and to determine the stability of its viral virulence.Methods The G protein genes of CTN-1V strain at passage 6 were amplified by RT-PCR.The products were cloned into pGEM-T vector and then were further sequenced.The DNAStar and Mega4.0 software were used for sequencing and homology analysis of glycoprotein gene.The virus titers of CTN-1V strain at passage 6 were calculated in LD50/ml and FFU/ml respectively,and the viral virulence was evaluated with FFU/LD50 index.Results The sequence analysis of glycoprotein genes of multiple passages of CTN-1V strain showed an only mutation from Guanine to Adenine at 1222 nt,leading to an amino acid change from Glu to Lys at 408 aa,but a nucleotide mutation from "A" to "G" at 1320 nt was nonsense.The virulence indices of CTN-1V strains at 5 passages were between 1.00 and 1.07.The Bioinformatics analysis showed that there were high homologies between CTN-1V strain and the street rabies virus isolated from different regions in China.Conclusion The glycoprotein (G) genes and the virulence of CTN-1V strain appeared to be stable during passaging.The CTN-1V strain showed high homologies with the street rabies viruses isolated from different regions in China.The results provided a supportive evidence for the better quality control of this virus strain as a reference for further research on rabies vaccine for human use in China.