河南农业科学
河南農業科學
하남농업과학
JOURNAL OF HENAN AGRICULTURAL SCIENCES
2009年
7期
114-119
,共6页
曹素芳%李明%王岩%刘长斗%朱赞梅%唐桂芬%肖松云
曹素芳%李明%王巖%劉長鬥%硃讚梅%唐桂芬%肖鬆雲
조소방%리명%왕암%류장두%주찬매%당계분%초송운
猪繁殖与呼吸综合征病毒%ORF5基因%克隆%序列分析
豬繁殖與呼吸綜閤徵病毒%ORF5基因%剋隆%序列分析
저번식여호흡종합정병독%ORF5기인%극륭%서렬분석
PRRSV%ORF5 gene%Clone%Genetic analysis
从河南信阳和新乡两地区猪场疑似猪繁殖与呼吸综合征病料中分离出能引起Marc145 细胞病变的病毒,命名为HN1和HN2.采用RT-PCR 方法从分离的2株PRRSV中均分别扩增出ORF5基因,并将其克隆、测序.用DNAStar软件分析所测序列,并与北美洲型VR-2332株及国内分离株进行核苷酸和推导的氨基酸同源性比较,绘制系统进化树,结果表明,ORF 5核苷酸与北美洲型的同源性为81.2%~99.1%,与欧洲型Lv株的同源性分别为35.9%和35.3%,推导氨基酸与北美洲型的同源性为84.5%~99.5%,与欧洲型Lv株的同源性分别为46.0%和45.5%.证明新分离到的PRRSV毒株仍属北美洲型.与国内分离的高致病性PRRSV毒株JXA1、HUB1、JX0612、GDZC1、ZQ、SX-1、HB1、HEB1、Hnyz核苷酸同源性达96.5%~99.1%,氨基酸同源性为96.0%~99.5%.分析结果表明,所获得的2个毒株GP5蛋白氨基酸序列中的抗原表位已经发生变异.
從河南信暘和新鄉兩地區豬場疑似豬繁殖與呼吸綜閤徵病料中分離齣能引起Marc145 細胞病變的病毒,命名為HN1和HN2.採用RT-PCR 方法從分離的2株PRRSV中均分彆擴增齣ORF5基因,併將其剋隆、測序.用DNAStar軟件分析所測序列,併與北美洲型VR-2332株及國內分離株進行覈苷痠和推導的氨基痠同源性比較,繪製繫統進化樹,結果錶明,ORF 5覈苷痠與北美洲型的同源性為81.2%~99.1%,與歐洲型Lv株的同源性分彆為35.9%和35.3%,推導氨基痠與北美洲型的同源性為84.5%~99.5%,與歐洲型Lv株的同源性分彆為46.0%和45.5%.證明新分離到的PRRSV毒株仍屬北美洲型.與國內分離的高緻病性PRRSV毒株JXA1、HUB1、JX0612、GDZC1、ZQ、SX-1、HB1、HEB1、Hnyz覈苷痠同源性達96.5%~99.1%,氨基痠同源性為96.0%~99.5%.分析結果錶明,所穫得的2箇毒株GP5蛋白氨基痠序列中的抗原錶位已經髮生變異.
종하남신양화신향량지구저장의사저번식여호흡종합정병료중분리출능인기Marc145 세포병변적병독,명명위HN1화HN2.채용RT-PCR 방법종분리적2주PRRSV중균분별확증출ORF5기인,병장기극륭、측서.용DNAStar연건분석소측서렬,병여북미주형VR-2332주급국내분리주진행핵감산화추도적안기산동원성비교,회제계통진화수,결과표명,ORF 5핵감산여북미주형적동원성위81.2%~99.1%,여구주형Lv주적동원성분별위35.9%화35.3%,추도안기산여북미주형적동원성위84.5%~99.5%,여구주형Lv주적동원성분별위46.0%화45.5%.증명신분리도적PRRSV독주잉속북미주형.여국내분리적고치병성PRRSV독주JXA1、HUB1、JX0612、GDZC1、ZQ、SX-1、HB1、HEB1、Hnyz핵감산동원성체96.5%~99.1%,안기산동원성위96.0%~99.5%.분석결과표명,소획득적2개독주GP5단백안기산서렬중적항원표위이경발생변이.
Two strains of virus causing CPE of Marc-145 cells were isolated from the tissues of dead pigs,which collected from Xinyang and Xinxiang areas of Henan province and primarily diagnosed as swine porcine reproductive and respiratory syndrome (PRRS).These two strains were named as HN1 and HN2, and the ORF5 genes of PRRSV were cloned and sequenced,respectively. Compared with those of VR-2332,Lv and other strains isolated in China, the ORF5 genes of HN1 and HN2 strains shared 81.2% to 99.1% nucleotide homologies with the North American type, and shared 35.9% and 35.3% with European Lv strain. The deduced amino acid homology shared 84.5% to 99.5% with North American type and shared 46.0% and 45.5% with European Lv strain. This indicated that the isolated strains were related to the genotype of North American PRRSV.The nucleotide sequence homologies were 96.5% to 99.1% and amino acid homologies were 96.0% to 99.5% comparing with the highly pathogenic PRRSV JXA1,HUB1,JX0612,GDZC1,ZQ,SX-1,HB1,HEB1,and Hnyz strains.The results showed that the antigen epitope of GP5 proteins of HN1 and HN2 strains have mutated.