广西狂犬病病毒L基因聚合酶活性部位的克隆与序列分析-预防兽医学专业论文.docxVIP

广西狂犬病病毒L基因聚合酶活性 广西狂犬病病毒L基因聚合酶活性 部位的克隆与序列分析 摘要 本实验对广西毒株狂犬病病毒L基因聚合酶活性部位进行克隆 和测序，截取L基因聚合酶活性部位核苷酸和推定的氨基酸序列与国 外固定毒株和类狂犬病毒株进行比较分析。 结果显示广西毒株与国外固定毒株的L基因聚合酶活性部位的 核苷酸和推定的氨基酸同源性分别为84．8％．87．5％和94．5％．99％， 与类狂犬病病毒株的核苷酸和推定的氨基酸同源性分别为75．5％ ．79．2％和93．5％．97％，广西各毒株之间的核苷酸和推定的氨基酸同 源性较高，分别为88．7％．99．8％和96．5％．100％。 根据L基因聚合酶活性部位的核苷酸序列对广西狂犬病毒株作 遗传进化分析，可以将广西毒株分成3个群，群I：GX08、GX09、 GX014、GX091、GXl95、GX260、GXHX、GXLA、GXSL、 GX、ⅣX； 群II：GX01、GX074、GX219、GX304、GXPX、GXBM为；群III- GXNll9。 比较了L基因聚合酶活性部位氨基酸的变异情况，广西毒株L基 因聚合酶活性部位的特有变异为第660位、745位和771位，其中在 L基因的第745位点上，广西毒株全部为精氨酸，而在该位点的国外 L基因的第745位点上，广西毒株全部为精氨酸，而在该位点的国外 参考毒株为丝氨酸、亮氨酸或组氨酸；在660位点上，广西毒株中 GX01、GX074、GX219、GX304、GⅪ7X和GXBM株为缬氨酸；771 位点上，GX08、GX09、GX014、GX091、GXl95、GX260、GXHX、 GXLA、GXSL和GXWX株为丙氨酸。 比较L基因聚合酶活性部位，发现其4个基序高度保守，其中基 序A保持不变；基序B、C、D只有少数氨基酸发生变异，基序C中 的核心序列GDNQ在各毒株中都不变。 关键词：狂犬病病毒聚合酶同源性序列分析 CLONING CLONING AND SEQUENCE ANAI YSIS OF L GENE POLYMERASE ACnVITY MODULE OF RABIES VIRUS ISOI。pdES FROM GUANGXI ABSTRACT In this study the eDNA fragments containing the L gene polymerase activity module of rabies virus isolates from Guangxi were doned and sequenced．The nucleotide sequence of L gene activity module and their deduced amino acid sequence were compared with those of other rabies virus strains and rabies-related virus strains published previously． Comparison of the nucleotide sequence and deduced amino acid sequences of polymerase activity module of rabies virus isolates from Guangxi with that of other fixed rabies virus strains and rabies—related virus strains．The nucleotide homologies and deduced amino acid homologies between isolates from Guangxi and fixed strains were 84．8％ 一87．5％and 94．5％一99％respectively,while Guangxi isolates and rabies—related strains were 75．5％-79．2％and 93．5％-97％，respectively． The nucleotide homologies and deduced amino acid homologies between Guangxi isolates were 88．7％一99．8％and 96．5％一100％respectively． According to the nucleotide sequence of L gene polymerase activity HI module，We module，We analyzed the homology identity