CN114957406B - 一种犬细小病毒的三价抗原和三价病毒样颗粒以及应用 - Google Patents
一种犬细小病毒的三价抗原和三价病毒样颗粒以及应用 Download PDFInfo
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- CN114957406B CN114957406B CN202210584627.0A CN202210584627A CN114957406B CN 114957406 B CN114957406 B CN 114957406B CN 202210584627 A CN202210584627 A CN 202210584627A CN 114957406 B CN114957406 B CN 114957406B
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- canine parvovirus
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Abstract
本发明公开一种犬细小病毒的三价抗原和三价病毒样颗粒以及应用，属于生物医学技术领域。为了提供一种安全性高，保护期长，免疫效果好的犬细小病毒的三价疫苗。本发明提供一种犬细小病毒的三价抗原是由犬细小病毒new CPV‑2a的VP2的氨基酸序列、犬细小病毒new CPV‑2b的VP2的氨基酸序列和犬细小病毒CPV‑2c的VP2的氨基酸序列组成的，以及提供病毒样颗粒的构建，制备犬细小病毒三价亚单位疫苗，为预防犬细小病毒病奠定基础，该方法显著提高产物表达量、抗原免疫原性高、保护范围广、成本低且具有规模化生产的优点。
Description
技术领域
本发明属于生物医药技术领域，具体涉及一种犬细小病毒的三价抗原和三价病毒样颗粒以及应用。
背景技术
犬细小病毒性肠炎是由犬细小病毒(Canine parvovirus,CPV)引起的一种急性、高度接触性传染性肠炎疾病，主要以呕吐剧烈、腹泻、粪便有恶臭味、出血性肠炎和白细胞数量减少为特征。
1978年第一次分离出CPV，被命名为CPV-2型。CPV-2型可感染犬但不能感染猫。1980年，Parrish等发现新的变异株CPV-2a，CPV-2a与CPV-2的VP2有5个氨基酸变异，导致病毒与单克隆抗体的反应特性、与猫转铁蛋白受体亲和性和猫体内复制能力均发生改变，CPV-2a逐渐取代了CPV-2在世界范围内流行。1984年，CPV-2b型被发现，CPV-2a和CPV-2b逐渐取代了CPV-2，CPV-2a和CPV-2b对犬的细胞更适应并且扩大了宿主的范围，不仅可以感染犬，在一定情况下还可以感染猫。2000年Ikeda等在豹猫中分离到一种新的细小病毒变异株，但不和CPV的另几种单抗反应，因此被认为该病毒是新的抗原型并命名为CPV-2c。CPV-2c比起CPV-2a和CPV-2b对猫的致病能力强。此外，还有new CPV-2a、new CPV-2b、CPV-2c(a)、CPV-2c(b)毒株陆续被发现，均是VP2关键氨基酸位点发生变异。我国目前主要是以newCPV-2a、new CPV-2b为主，偶发CPV-2c，这提示CPV在长期进化和免疫压力下已经发生了变异，这可能也是临床上CPV感染病例逐渐增多和免疫犬发病的重要原因之一。
犬细小病毒病没有有效的治疗办法，主要依靠接种疫苗进行免疫预防。现有疫苗为弱毒疫苗和灭活疫苗。弱毒苗免疫犬后，病毒能在犬体内增殖，可能将疫苗毒排出传递给未免疫的犬，或毒株突变、毒力返强而爆发疾病；灭活疫苗由于病毒的核酸已被破坏，使得病毒已不具有感染性，所以其安全性高且不存在散毒的风险，但免疫持续期短。且目前商品化的疫苗只有以CPV-2型或CPV-2b型病毒基础上制备的，均为弱毒疫苗，这导致国内流行毒株与疫苗毒株不匹配，且现有毒株疫苗对新型突变株的保护性明显降低，国内外均有犬群接种疫苗后仍暴发CPV的报道。因此，制备新型CPV疫苗的呼声渐长，以流行毒株分离株为疫苗用毒研制出新型的犬细小病毒三价疫苗是非常必要的。
发明内容
本发明的目的是为了提供一种安全性高，保护期长，免疫效果好的犬细小病毒的三价疫苗。
本发明提供一种犬细小病毒的三价抗原，所述三价抗原是由犬细小病毒new CPV-2a的VP2的氨基酸序列、犬细小病毒new CPV-2b的VP2的氨基酸序列和犬细小病毒CPV-2c的VP2的氨基酸序列组成的。
进一步地限定，所述犬细小病毒new CPV-2a的VP2的氨基酸序列如SEQ ID NO.18所示；所述犬细小病毒new CPV-2b的VP2的氨基酸序列如SEQ ID NO.19所示；所述犬细小病毒CPV-2c的VP2的氨基酸序列如SEQ ID NO.20所示。
进一步地限定，犬细小病毒new CPV-2a的VP2的基因序列如SEQ ID NO.5所示；犬细小病毒new CPV-2b的VP2的基因序列如SEQ ID NO.6所示；犬细小病毒CPV-2c的VP2的基因序列如SEQ ID NO.7所示。
本发明提供一种重组杆状病毒，其特征在于，所述重组杆状病毒是以E.coli DH10Bac为出发菌，表达上述三价抗原获得的重组微生物，然后利用重组微生物感染Sf9细胞、Sf21细胞或High Five细胞获得的。
本发明提供一种犬细小病毒的三价病毒样颗粒，其特征在于，所述三价病毒样颗粒是利用上述的重组杆状病毒感染Sf9细胞、Sf21细胞或High Five细胞，经过离心后收集细胞培养物经过细胞裂解处理后获得的。
本发明提供上述的三价病毒样颗粒的制备方法，其特征在于，所述方法如下：
步骤1：将CP-2A-opt基因序列、CP-2B-opt基因序列和CP-2C-opt基因序列连接至转移载体pFF获得重组载体pFF-CP-ABC；所述CP-2A-opt基因序列如SEQ ID NO.2所示；CP-2B-opt基因序列如SEQ ID NO.3所示；CP-2C-opt基因序列如SEQ ID NO.4所示；
步骤2：将重组载体pFF-CP-ABC利用热休克方法转化E.coli DH10 Bac感受态获得重组杆状病毒Bac-CPV-ABC；
步骤3：将重组杆状病毒Bac-CPV-ABC转染贴壁Sf9细胞，收集细胞培养物，离心10min，分离上清液和细胞，细胞加入NaHCO3悬浮处理后，离心后获取上清液。
进一步地限定，步骤1中获得转移载体pFF的方法如下：
步骤1.1：pFastBacDual载体p10启动子下的MCS中，将Kpn I至Nhe I的序列删除；将pH启动子下MCS中的Rsr II至Spe I的序列删除获得质粒pFD(k+n)(r+s)，将基因序列10H通过BstZ17 I酶切位点连入质粒pFD(k+n)(r+s)；
步骤1.2：将合成的序列10H的基因序列如SEQ ID NO.1所示，通过BstZ17 I酶切位点连入质粒pFD(k+n)(r+s)获得转移载体pFF，转移载体pFF的序列如SEQ ID NO.21所示。
本发明一种犬细小病毒的三价疫苗，所述三价疫苗由上述的三价病毒样颗粒和Montanide Gel 02PR佐剂组成。
本发明提供上述的三价抗原或上述的三价疫苗在制备预防或治疗犬细小病毒性肠炎的疫苗或药物中的应用。
本发明提供上述述的三价抗原或上述的三价病毒样颗粒在制备犬细小病毒性肠炎的三价卵黄抗体中的应用。
有益效果：利用分子生物学技术对new CPV-2a、new CPV-2b、CPV-2c型流行毒株进行病毒样颗粒的构建，制备犬细小病毒三价亚单位疫苗，免疫犬细小三价疫苗后可在犬血清中可检测针对new CPV-2a、new CPV-2b、CPV-2c株的HI效价。加强免疫后，抗体效价进一步升高，在免疫后3、4周达到最高，均不低于1:212。
附图说明
图1为pFF载体示意图；
图2为转移载体构建模式图；
图3为PCR图，其中，M：Marker 2000；1：CP-A-opt；2：CP-B-opt；3：CP-C-opt；
图4为转移载体pFF-CP-ABC的酶切鉴定图，其中，M：Marker 15000；1：Spe I+Mlu I双酶切鉴定；2：Not I+Hind III双酶切鉴定；3：Sma I+Xho I双酶切鉴定；
图5为重组bacmid的PCR鉴定图，其中，M：Marker 15000；1：p10表达框引物鉴定图；2：pH表达框引物鉴定图；
图6为重组病毒感染Sf9细胞的形态(100×)，其中，A为CPV-ABC株感染的Sf9细胞；B为正常Sf9细胞；
图7为CPV抗原检测试纸条；
图8为免疫荧光染色鉴定VP2表达(100×)，其中，A为重组杆状病毒Bac-CPV-ABC株感染的Sf9细胞；B为空白对照Sf9细胞；
图9为SDS-PAGE鉴定，其中，1为细胞培养物，M为预染Marker，2为细胞处理并离心后的上清；
图10为电镜负染观察病毒样颗粒；
图11为CPV HI抗体效价，其中，横坐标为免疫天数，纵坐标为CPV HI抗体效价。
具体实施方式
new CPV-2a、new CPV-2b、CPV-2c(记载在文章Buonavoglia D C.Canineparvovirus—A review of epidemiological and diagnostic aspects,with emphasison type 2c[J].Veterinary Microbiology,2012.)
实施例1.pFastBacDual载体改造
将pFastBacDual进行载体改造，获得含有4个多克隆酶切位点(MCS)的转移载体。
1.序列合成
选取pFastBacDual载体3960-4989区域，包含p10启动子、MCS、HSV tk多聚腺苷酸化信号、pH启动子、MCS、SV40多聚腺苷酸化信号，并进行一系列改动：在4644nt处，引入序列GCTAAGCTAGATCGAT；将4461nt的C改为G；将p10处MCS，从Bbs II至Nco I删除，并引入Pme I；将pH处MCS，从Not I至Hind III删除，并引入Mlu I。将改动后的序列拼接，命名为10H(基因序列如SEQ ID NO.1所示)，送至上海生工合成在pUC载体上。
2.pFastBacDual载体酶切位点删除
pFastBacDual载体p10启动子下的MCS中，将Kpn I至Nhe I的序列删除；将pH启动子下MCS中的Rsr II至Spe I的序列删除，方法如下：
将pFastBacDual载体经KpnI+NheI双酶切处理，酶切体系50μl组成：模板2.5μg，Kpn I和Nhe I各2.5μl，10×Green Buffer 5μl，补dd H2O至50μl。混匀后，在37℃条件下酶切3h。然后，将纯化后的酶切载体使用Kellow大片段进行处理，体系组成：模板25ng，10×Kellow Buffer 2.5μl，dNTP 2.5μl，Kellow 1μl，补dd H2O至25μl。混匀后，在37℃条件下反应3h，65℃5min。再使用T4连接酶连接，连接体系5μl组成：T4 buffer 0.5μl，T4连接酶0.5μl，模板4μl，混匀后，20℃连接5h。最后，将连接产物与感受态细胞(E.coli DH5a)采用热休克方法进行转化，筛选获得质粒pFD(k+n)。用同样方法，将pFD(k+n)经Rsr II+Spe I处理、连接，获得质粒pFD(k+n)(r+s)。
3.获得拼接pFF载体
将合成的序列10H(SEQ ID NO.1)通过BstZ17 I酶切位点连入质粒pFD(k+n)(r+s)中，酶切体系50μl组成：模板2.5μg，BstZ17 I 2.5μl，10×Green Buffer 5μl，补dd H2O至50μl。混匀后，在37℃条件下酶切3h。将纯化后的载体、片段使用T4连接酶连接，连接体系5μl组成：T4 buffer 0.5μl，T4连接酶0.5μl，载体0.5μl，片段3.5μl，混匀后，20℃连接5h。将连接产物与感受态细胞(E.coli DH5a)采用热休克方法进行转化，筛选获得转移载体pFF。转移载体pFF有4套启动子、相应的多克隆酶切位点和多聚腺苷酸化信号，以便于外源片段的连入，分别命名为：p101、p102、pH1、pH2。其中，p101、pH2启动子分别为pFastBacDual载体的p10、pH启动子，p102、pH1启动子分别为通过合成序列10H引入载体的p10、pH启动子。转移载体pFF的载体示意图见图1，转移载体pFF的序列如SEQ ID NO.21所示。
实施例2.重组杆状病毒的构建与鉴定
1.引物设计
将分离CPV流行毒株时获得的new CPV-2a(new CPV-2a的VP2基因序列如SEQ IDNO.5所示)、new CPV-2b(new CPV-2b的VP2基因序列如SEQ ID NO.6所示)、CPV-2c(CPV-2c的VP2基因序列如SEQ ID NO.7所示)株的VP2序列根据密码子偏爱性优化，优化后的序列命名为CP-2A-opt(SEQ ID NO.2)、CP-2B-opt(SEQ ID NO.3)、CP-2C-opt(SEQ ID NO.4)，由上海生工合成。根据转移载体pFF的MCS，设计引物，见表1。
表1引物信息
2.转移载体的构建与鉴定
将CP-2A-opt、CP-2B-opt、CP-2C-opt序列分别连接至转移载体pFF的表达盒中，示意图见图2。先通过PCR获得CP-2A-opt、CP-2B-opt、CP-2C-opt的PCR片段，引物见表1，PCR产物见图3。再将pFF和CP-2A-opt VP2 PCR产物分别进行双酶切处理，酶切体系50μl组成：模板2.5μg，Spe I和Mlu I各2.5μl，10×Green Buffer 5μl，补dd H2O至50μl。混匀后，在37℃条件下酶切3h。然后，将纯化后的VP2片段连入双酶切处理的载体中，连接体系5μl组成：T4连接酶buffer 0.5μl，T4连接酶0.5μl，片段3.5μl，载体0.5μl，混匀后，20℃连接5h。最后，将连接产物与感受态细胞(E.coli DH5a)采用热休克方法进行转化，获得阳性质粒pFF-CP-A。同样，按照上述方法通过酶切连接将CP-2B-opt VP2 PCR产物基因连接至pFF-CP-A载体的Not I+Hind III酶切位点，获得pFF-CP-AB；将CP-2C-opt VP2 PCR产物基因连接至pFF-CP-AB载体的Sma I+Xho I酶切位点，获得pFF-CP-ABC。采用核酸内切酶对pFF-CP-ABC双酶切鉴定，均可获得约为1755bp和9663bp左右的两个片段，符合预期片段大小，见图4所示。
3.获得重组杆状病毒基因组(bacmid)
将转移载体pFF-CP-ABC利用热休克方法转化E.coli DH10 Bac感受态，条件如下：将感受态细胞从-70℃冰箱内取出至冰上融化，加入2μl pFF-CP-ABC混匀后，冰浴30min，42℃热应激50s，再冰浴3min，加入1ml无抗性LB培养基，在37℃200r/min条件下培养4h，菌液做10、100、1000倍比连续稀释，从每种稀释液中分别取出100μl涂布于含有三抗抗性细菌培养板上(含50μg/ml卡那霉素、7μg/ml庆大霉素、10μg/ml四环霉素、100μg/ml X-gal和40μg/ml IPTG)，37℃培养48h后，进行蓝白斑筛选。挑取白斑菌落，划线于三抗抗性细菌培养板上，继续培养48h之后，挑取白色菌落，接菌至含三抗抗性的液体LB培养基中，在37℃200r/min条件下培养12～16h之后，采用碱裂解与异丙醇沉淀结合的方法提取重组bacmid：Bac-CPV-ABC。根据pFF设计bacmid鉴定引物，见表2。以bacmid鉴定引物对Bac-CPV-ABC进行PCR鉴定，结果见图5，扩增产物均为阳性片段，与预期大小相符。以上结果表明外源基因片段正确重组到杆状病毒基因组的目的位点上。
表2 bacmid鉴定引物信息
犬细小病毒new CPV-2a的VP2的氨基酸序列如SEQ ID NO.18所示；所述犬细小病毒new CPV-2b的VP2的氨基酸序列如SEQ ID NO.19所示；所述犬细小病毒CPV-2c的VP2的氨基酸序列如SEQ ID NO.20所示。
实施例3.重组杆状病毒的获得
将提取的bacmid Bac-CPV-ABC转染贴壁Sf9细胞，转染过程如下：将2μg重组DNA加入到100μl和双抗的TNM-FH培养液中，混匀。将9μl Cellfectin Reagent加入到100μl TNM-FH培养液中，混匀。将重组DNA与脂质体混合，室温静止40min。将用6孔板培养好的昆虫细胞Sf9(覆盖孔底面积已经达到80％～90％)用TNM-FH培养液轻洗三次，然后，每孔加入1mlTNM-FH培养液，待备用。将重组DNA与脂质体的混合物轻轻加入到每孔细胞中，轻轻混匀，在27℃条件下静止培养5～6h。将孔中的液体弃掉，加入2ml完全TNM-FH培养液，27℃静止培养5～6天，待细胞肿胀，体积变大，脱落后，收集上清液，命名为CPV-ABC株P1代。用其感染新培养的Sf9悬浮细胞，连续传代2代后，收取细胞上清液，4℃或-70℃保存备用，获得重组杆状病毒CPV-ABC。
将重组杆状病毒CPV-ABC株感染贴壁培养的Sf9昆虫细胞，将细胞置于27℃温箱内静止培养。
接毒后2～4天观察细胞变化，正常未接毒的细胞为大小均一、折光性良好、轮廓清晰的透明球形细胞(见图6B)，而重组病毒感染的Sf9昆虫细胞出现明显的细胞病变，表现为：细胞体积明显增大、细胞脱落、细胞内颗粒增多(见图6A)。
实施例4.犬细小病毒三价病毒样颗粒
蛋白液的获取方法：将重组杆状病毒CPV-ABC株以MOI＝0.1感染悬浮的Sf9细胞，培养96h之后，收集细胞培养物，5000r/min离心10min，分离上清液和细胞，细胞加入与原收获细胞培养物等体积的25mM NaHCO3悬浮后，至冰上30min。5000r/min离心10min，获取上清液，即为犬细小病毒三价病毒样颗粒。
利用以下实验验证实验效果：
(1)试纸条检测：取犬细小病毒三价病毒样颗粒，使用犬细小抗原检测试纸条初步检测是否为细小病毒VP2蛋白。即将样品50倍稀释后，滴入加样孔内，5min后看结果。
结果表明(图7)，C线(对照线)有条带，说明试纸条有效；T线(检测线)处出现较强的阳性反应，表明重组杆状病毒CPV-ABC株可成功表达VP2蛋白。
(2)HA检测：取猪红细胞于阿氏液中保存，用PBS(0.015mol/L，pH 6.5)洗涤红细胞三次，最后将红细胞用PBS配成1％悬液，再加入0.5％兔血清。在V形板的每孔加入灭菌的PBS 25μl；用微量移液器取待测样品25μl加入第1孔内，混匀后，吸取25μl后加入到第二孔中，如此连续稀释至第23孔，第23孔吸取25μl液体弃掉；第24孔为PBS对照，阴性细胞培养物按相同方法稀释。每孔补加25μl PBS和50μl 1％猪红细胞悬液，混匀后，4℃下反应60min，待对照的红细胞完全沉积后观察结果。
结果显示，收获的犬细小病毒三价病毒样颗粒血凝效价可达到1：219，而new CPV-2a、new CPV-2b毒株、CPV-2c毒株感染哺乳动物细胞(F81细胞)培养获得的抗原HA效价均为1：210。表明制备的犬细小病毒三价病毒样颗粒保持了凝集猪红血球的血凝特性，并且表达量远大于哺乳动物细胞培养收获的抗原量。
(3)间接免疫荧光鉴定：采用间接免疫荧光试验检测VP2蛋白表达情况。取重组杆状病毒CPV-ABC株，将毒种用细胞培养液作20倍稀释，接种已长满80％单层的96孔Sf9细胞培养板，接种3孔，每孔50μl，同时设正常细胞对照。在27℃条件下，细胞静止培养48h之后，吸掉上清液，每孔加入100μl 80％丙酮溶液，在室温条件下，固定Sf9细胞30min，用PBST溶液洗涤3次后，细胞与犬细小病毒阳性血清(兔源)(1:200倍稀释)37℃反应1h；用PBST溶液洗涤3次后，加入FITC标记的羊抗兔IgG 37℃反应1h，用PBST溶液洗涤3次后，使用荧光显微镜检测结果。
实验结果显示，重组病毒感染的Sf9细胞展现出高强度的黄绿色荧光(见图8A)；对照组细胞无可见绿色荧光(见图8B)，表明重组杆状病毒CPV-ABC株感染Sf9细胞可表达CPVVP2蛋白。
(4)SDS-PAGE检测：将细小病毒三价病毒样颗粒取100μl加入25μl 5×上样缓冲液混匀，煮沸5min，用于SDS-PAGE检测。
结果显示，收获的细胞用NaHCO3处理后，目的蛋白可以分泌到上清中，获得的目的蛋白集中位于65kDa左右(见图9)。
(5)电镜观察：将细小病毒三价病毒样颗粒吸附到金属网上，用1％磷钨酸染色2～3min，然后，用滤纸吸掉金属网上的残余染色液，最后，利用电镜(JEM 1200EXII)观察。
电镜结果显示，从目的蛋白样品中可见到圆形球状病毒样颗粒(VLPs)(图10)，直径大小在20nm左右，其形态结构与天然CPV相似。表明重组杆状病毒CPV-ABC株感染的Sf9细胞能够正确表达CPV VP2蛋白，而且，表达的VP2蛋白能够装配成完整的CPV VLPs。
实施例5.犬细小病毒三价疫苗的制备与免疫原性评价
1.疫苗制备
将实施例4中制备的犬细小病毒病毒样颗粒与Montanide Gel 02PR佐剂按7:1比例混匀，制备成犬细小病毒三价疫苗，2～8℃保存备用。
2.犬免疫实验
用4～12周龄健康易感犬5只(CPV HI抗体效价不高于1:8)，每只皮下注射犬细小病毒三价疫苗1ml，21日后加强免疫一次。于首免后0、21、28、35、42日采血，分离血清，测定血清中犬细小病毒HI抗体效价。
3.犬细小病毒HI抗体测定
将采集的血清经56℃灭活30min，加入1/10体积猪红血球混匀，2～8℃吸附2h，3000r/min离心5min，取上清。在96孔V型微量血凝反应板的1～16孔加入PBS 25μl，然后在第1孔加入待检血清25μl，混匀，吸取25μl混合液加入第2孔，混匀，依次2倍系列稀释至第14孔，从第14孔吸取25μl弃去。在第1孔至第15孔每孔加入8单位new CPV-2a(CPV-SD06株经过鉴定是属于new CPV-2a毒株)、new CPV-2b(CPV-SY02株经过鉴定是属于new CPV-2b毒株)或CPV-2c(CPV-SY05株经过鉴定是属于CPV-2c毒株)25μl，第15孔作抗原对照，第16孔加入PBS 25μl作红细胞对照。振荡混匀后，37℃作用60min。每孔加入1％猪红细胞悬液(含0.5％兔血清)50μl，混匀，2～8℃作用60min，观察HI效价。
免疫结果表明，免疫犬细小三价疫苗后可在犬血清中可检测针对new CPV-2a、newCPV-2b、CPV-2c株的HI效价(图11)。加强免疫后，抗体效价进一步升高，在免疫后3、4周达到最高，均不低于1:212。实验结果表明，制备的犬细小三价疫苗有良好的免疫原性。
4.攻毒保护实验
将免疫犬分为6组，分为A组、B组、C组、D组、E组、F组，每组5只。A组、B组、C组皮下免疫犬细小三价疫苗，D组、E组、F组皮下免疫MEM培养液，剂量为1.0ml/只。免疫后21天加强免疫一次。加强免疫后21天进行攻毒，将A组、D组攻毒CPV-SD06株，将B组、E组攻毒CPV-SY02株，将C组、F组攻毒CPV-SY05株，口服毒株15ml/只(血凝素凝集价1：29)。观察记录试验动物的发病情况和死亡情况，连续记录9天。计算疫苗组和发病对照组的发病率，以评估制品的效力。
结果显示，D组、E组、F组对照组在攻毒后，均出现不同程度的精神状态差、食欲减退、呕吐、血便等临床症状，发病率100％。A组、B组、C组免疫组试验犬均未出现临床症状发病率均为0％，保护率为100％。说明制备的犬细小病毒三价疫苗具有良好的免疫攻毒保护力。
5.安全性实验
对健康幼犬(4～12周龄)进行一次单剂量注射，一次单剂量重复注射和一次超剂量注射，均安全性良好。对部分免疫的幼犬进行解剖学和组织学检查，均未出现肉眼可见的病理变化。表明犬细小三价疫苗灭活疫苗安全性良好。
实施例6.犬三价卵黄抗体的制备
1.疫苗制备：利用实施例4获得的犬细小病毒三价病毒样颗粒与佐剂(弗氏完全佐剂或弗氏不完全佐剂)1:1(v：v)混合，用乳化器进行乳化，乳化至液体滴入水中不化开为宜。将乳化好的疫苗装入疫苗瓶中，备用。
2.免疫鸡
选取20只90～110日龄健康海兰白蛋鸡，随机分为两组，一组为免疫组，一组为空白对照组。使用连续注射器进行免疫，腿部肌肉注射，14天加强免疫一次。首次免疫时，免疫组使用弗氏完全佐剂乳化制备的疫苗进行免疫，1ml/只；第2～4次免疫时，使用弗氏不完全佐剂乳化制备的疫苗进行免疫，分别为1.5ml/只、2ml/只和2ml/只。
3.卵黄抗体制备与效价测定：第3次免疫、第4次免疫后第14天收集鸡蛋，分离蛋黄，使用50mM乙酸-乙酸钠缓冲液进行10倍稀释后，检测犬细小病毒HI抗体效价。
结果表明，在第三次免疫后2周(W6)，免疫组均有HI抗体效价产生。且随着免疫次数的增加，抗体效价逐渐升高，且均可产生针对犬细小病毒三种型别的HI抗体(表3)。表明使用犬细小三价抗原可成功制备CPV三价卵黄抗体。
表3 CPV HI抗体效价
SEQUENCE LISTING
<110> 吉林大学；长春西诺生物科技有限公司
<120> 一种犬细小病毒的三价抗原和三价病毒样颗粒以及应用
<160> 21
<170> PatentIn version 3.5
<210> 1
<211> 990
<212> DNA
<213> 人工合成
<400> 1
tatagttcta gtggttggcc tacgtacccg tagtggctat ggcagggctt gccgccccga 60
cgttggctgc gagccctggg ccttcacccg aacttggggg ttggggtggg gaaaaggaag 120
aaacgcgggc gtattggtcc caatggggtc tcggtggggt atcgacagag tgccagccct 180
gggaccgaac cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt 240
tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc 300
ctcccccatc tcccgtttaa acggtaccgc atgctatgca tcagctgcta gcagtcttcg 360
tcgagtgatt gtaaataaaa tgtaatttac agtatagtat tttaattaat atacaaatga 420
tttgataata attcttattt aactataata tattgtgttg ggttgaatta aaggtccgta 480
tagtcgctaa gctagatcga tcggaatatt aatagatcat ggagataatt aaaatgataa 540
ccatctcgca aataaataag tattttactg ttttcgtaac agttttgtaa taaaaaaacc 600
tataaatatt ccggattatt cataccgtcc caccatcggg cgcggatccc ggtccgaagc 660
gcgcggaatt caaaggccta cgtcgacgag ctcactagta cgcgtgtcga gaagtactag 720
aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca 780
cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 840
gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 900
ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 960
atctgatcac tgcttgagcc taggagatcc 990
<210> 2
<211> 1755
<212> DNA
<213> 人工合成
<400> 2
atgagtgacg gtgccgtgca gcctgatggc ggccagccag ctgtgcgtaa cgagcgcgcc 60
accggtagcg gtaacggcag cggcggtgga ggcggcggcg gtagcggtgg tgtgggcatc 120
tccaccggca cattcaacaa ccagaccgag ttcaagtttc tggagaacgg ttgggtggaa 180
atcaccgcta actccagccg cctcgtccac ctgaacatgc cagaatcaga aaactaccgc 240
cgtgtggtgg tgaacaacct ggacaagact gccgtgaacg gcaacatggc cctggacgac 300
acccacgccc agatcgtcac tccatggagc ctggtggacg ctaacgcttg gggcgtttgg 360
ttcaaccctg gtgactggca gctgatcgtg aacaccatgt ctgaactgca cctggtgagc 420
ttcgagcagg aaatcttcaa cgtggtgctg aagacagtga gcgaatcagc cactcagcca 480
cctacaaagg tgtacaacaa cgacctgacc gctagcctga tggtggccct cgacagcaac 540
aacaccatgc ccttcacccc tgccgctatg cgtagcgaga ctctcggctt ctacccatgg 600
aagcctacca tccctacccc ttggcgctac tacttccagt gggaccgcac cctgatcccc 660
tcccacacag gtacatccgg tacccccacc aacatctacc acggcaccga ccctgacgac 720
gtgcagttct acaccatcga aaacagcgtg cccgtgcacc tgctgcgcac tggtgacgag 780
ttcgccactg gcacattcta cttcgactgt aagccctgcc gcctgaccca cacctggcag 840
actaaccgtg ccctgggcct gcctcctttc ttgaacagcc tcccccaggc tgaaggaggt 900
actaacttcg gctacatcgg cgtgcagcag gacaagcgtc gtggtgtgac ccagatggga 960
aacactaaca tcatcaccga agccaccatc atgcgtcctg ccgaggtggg ctacagcgct 1020
ccctactact ccttcgaggc tagcacccag ggcccattca agacccctat cgccgccggc 1080
cgcggcggtg cccagacaga cgagaaccag gctgctgacg gcgaccctcg ttacgctttc 1140
ggccgtcagc acggccagaa gaccacaaca accggtgaaa cccctgaacg cttcacttac 1200
atcgctcacc aggacactgg caggtaccct gagggtgact ggatccagaa catcaacttc 1260
aacctgcctg tgaccaacga caacgtgctc ctccccactg acccaatcgg cggcaaggcc 1320
ggtatcaact acactaacat cttcaacaca tacggccccc tgacagccct gaacaacgtc 1380
ccacccgtct accccaacgg ccagatctgg gacaaggagt tcgacaccga cctgaagccc 1440
cgcctgcacg tcaacgctcc tttcgtgtgc cagaacaact gtcccggcca gctcttcgtg 1500
aaggtggccc ctaacctgac aaacgagtac gacccagacg ctagcgctaa catgtctcgt 1560
atcgtgacat actccgactt ctggtggaag ggcaagctgg tgttcaaggc taagctgcgt 1620
gcctcacaca cttggaaccc tatccagcag atgagcatca acgtggacaa ccagttcaac 1680
tacgtgccta gcaacatcgg tggtatgaag atcgtttacg aaaagtccca gctcgctcca 1740
cgtaagctgt actaa 1755
<210> 3
<211> 1755
<212> DNA
<213> 人工合成
<400> 3
atgtctgacg gtgctgtgca gcctgacggc ggtcagccag ctgtgcgtaa cgaacgcgcc 60
actggtagcg gcaacggctc cggtggaggt ggcggcggcg gtagcggcgg tgtgggcatc 120
agcaccggca ccttcaacaa ccagactgaa ttcaagttcc tcgaaaacgg ctgggttgaa 180
atcaccgcca actcctctcg tctggtgcac ctgaacatgc cagagagcga gaactaccgt 240
cgtgtggtgg tgaacaacct ggataagact gctgtgaacg gtaacatggc tctggacgac 300
acccacgctc agatcgtgac cccttggtcc ctggttgacg ctaacgcctg gggagtgtgg 360
ttcaaccctg gtgactggca gctgatcgtg aacaccatgt ccgagctgca cctggtgtcc 420
ttcgaacagg aaatcttcaa cgtggtgctg aagaccgtgt ccgaaagcgc cactcagccc 480
cctactaagg tctacaacaa cgacctgacc gcttccctga tggttgctct ggacagcaac 540
aacactatgc ctttcacccc cgctgctatg cgctccgaga cactcggctt ctacccatgg 600
aagcctacca tccccactcc ctggcgctac tacttccagt gggaccgtac cctgatccca 660
tcccacactg gcacttccgg taccccaact aacatctacc acggcactga cccagacgac 720
gtccagttct acacaatcga aaacagcgtg ccagtgcacc tgctgcgtac tggtgacgag 780
ttcgccaccg gaactttcta cttcgactgt aagccttgtc gcctgaccca cacatggcag 840
acaaaccgtg ctctgggact gccccccttc ctgaacagcc tgcctcaggc tgaaggtggt 900
acaaacttcg gttacatcgg cgtgcagcag gacaagcgtc gcggtgtgac ccagatgggt 960
aacactaaca tcatcaccga agccactatc atgcgccctg ccgaagtggg ttacagcgct 1020
ccctactaca gcttcgaagc ttccactcag ggtccattca agacacctat cgctgccggt 1080
aggggtggtg cccagacaga cgagaaccaa gccgctgacg gtgaccctcg ctacgccttc 1140
ggccgccagc acggccagaa gaccactacc acaggtgaga cccctgagcg tttcacttac 1200
atcgcccacc aagacacagg ccgctaccct gagggcgact ggatccagaa catcaacttc 1260
aacctccccg tgaccgacga caacgtgctg ctgcctaccg accccatcgg aggaaaggcc 1320
ggcatcaact acacaaacat cttcaacacc tacggcccac tcacagctct caacaacgtg 1380
ccacccgtgt accctaacgg ccagatctgg gacaaggagt tcgacaccga cctcaagcca 1440
cgcctgcacg tgaacgcccc tttcgtttgc cagaacaact gtcctggcca gctgttcgtg 1500
aaggtggccc ctaacctcac taacgaatac gaccctgacg ctagcgctaa catgtcccgc 1560
atcgtgacct actctgactt ctggtggaag ggcaagctcg tgttcaaggc caagctccgc 1620
gcttcccata cctggaaccc tatccagcaa atgtcaatca acgttgataa ccaattcaac 1680
tacgttccct ccaacatcgg cggtatgaag atcgtctacg aaaagagtca actggctcca 1740
cgtaagctgt actaa 1755
<210> 4
<211> 1755
<212> DNA
<213> 人工合成
<400> 4
atgagcgacg gtggtgtgca acctgacggc ggtcagcctg ctgtgcgtaa cgaacgtgct 60
actggtagcg gcaacggcag tggaggcggc ggcggtggcg gctccggcgg tgtgggcatc 120
tccaccggca ctttcaacaa ccagaccgaa ttcaagttcc tggagaacgg ttgggtggag 180
atcacagcta actccagccg cctggttcac ctgaacatgc ctgagtcaga aaactatcgt 240
cgcgtcgtcg tgaacaacct cgacaagacc gccgtgaacg gtaacatggc tctggacgac 300
acccacgccc agatcgtgac cccctggtca ctcgtcgacg ccaacgcttg gggtgtgtgg 360
ttcaacccag gcgactggca attgatcgtt aataccatgt cagagctgca cctcgtgtca 420
ttcgaacagg aaatcttcaa cgtggtgctg aagaccgtga gcgagtccgc tacccagccc 480
cctaccaagg tgtacaacaa cgacctgacc gctagcctga tggtggccct ggactccaac 540
aacaccatgc ctttcacacc cgccgccatg cgctccgaaa ccctgggttt ctacccctgg 600
aagccaacca tccctacccc ctggcgctac tacttccagt gggaccgtac cctgatccct 660
agccacactg gtacctccgg cactcccact aacatctacc acggcactga ccccgacgac 720
gtccagttct acaccatcga aaactccgtg cccgtccacc tgctgcgcac cggcgacgag 780
ttcgccaccg gcactttcta cttcgactgc aagccatgcc gcctcacaca cacttggcaa 840
acaaacagag ccctgggcct cccccctttc ctgaactccc tccctcaagc tgaaggtgga 900
acaaacttcg gatacatcgg agtgcagcag gacaagcgcc gcggcgtcac ccagatgggc 960
aacaccaaca tcatcaccga agctaccatc atgcgcccag ctgaagtggg ctacagcgcc 1020
ccctactact cattcgaggc ctccactcag ggccctttca agacccctat cgctgccggc 1080
cgtggtggcg ctcagaccga cgagaaccgt gctgctgacg gtgaccctcg ctacgctttc 1140
ggacgccagc acggccagaa gacaaccaca acaggtgaga cccctgagcg tttcacctac 1200
atcgcccacc aggacaccgg tcgctaccca gaaggtgact ggatccagaa catcaacttc 1260
aacctgccag tcacagagga caacgtgctg ctgcctaccg accccatcgg cggcaagacc 1320
ggtatcaact acaccaacat cttcaacacc tacggccctc tgacagccct gaacaacgtc 1380
ccacctgtgt accctaacgg tcagatctgg gacaaggagt tcgacaccga cctcaagcca 1440
cgtctgcacg tgaacgctcc tttcgtgtgt cagaacaact gtccaggcca gctgttcgtg 1500
aaggttgccc ctaacctgac taacgagtac gacccagacg cttccgctaa catgtcccgt 1560
atcgtgacat acagcgactt ctggtggaag ggcaagctgg tgttcaaggc caagctccgt 1620
gcctctcaca cctggaaccc tatccagcag atgtccatca acgtggacaa ccagttcaac 1680
tacgtgccta gcaacatcgg cggcatgaag atcgtgtacg aaaagtccca gctggcccct 1740
cgtaagctgt actaa 1755
<210> 5
<211> 1755
<212> DNA
<213> 人工合成
<400> 5
atgagtgatg gagcagttca accagacggt ggtcagcctg ctgtcagaaa tgaaagagct 60
acaggatctg ggaacgggtc tggaggcggg ggtggtggtg gttctggggg tgtggggatt 120
tctacgggta ctttcaataa tcagacggaa tttaaatttt tggaaaacgg atgggtggaa 180
atcacagcaa actcaagcag acttgtacat ttaaatatgc cagaaagtga aaattataga 240
agagtggttg taaataattt ggataaaact gcagttaacg gaaacatggc tttagatgat 300
acccatgcac aaattgtaac accttggtca ttggttgatg caaatgcttg gggagtttgg 360
tttaatccag gagattggca actaattgtt aatactatga gtgagttgca tttagttagt 420
tttgaacaag aaatttttaa tgttgtttta aagactgttt cagaatctgc tactcagcca 480
ccaactaaag tttataataa tgatttaact gcatcattga tggttgcatt agatagtaat 540
aatactatgc catttactcc agcagctatg agatctgaga cattgggttt ttatccatgg 600
aaaccaacca taccaactcc atggagatat tattttcaat gggatagaac attaatacca 660
tctcatactg gaactagtgg cacaccaaca aatatatacc atggtacaga tccagatgat 720
gttcaatttt acactattga aaattctgtg ccagtacact tactaagaac aggtgatgaa 780
tttgctacag gaacatttta ttttgattgt aaaccatgta gactaacaca cacatggcaa 840
acaaatagag cattgggctt accaccattt ctaaattctt tgcctcaagc tgaaggaggt 900
actaactttg gttatatagg agttcaacaa gataaaagac gtggtgtaac tcaaatggga 960
aatacaaaca ttattactga agctactatt atgagaccag ctgaggttgg ttatagtgca 1020
ccatattatt cttttgaggc gtctacacaa gggccattta aaacacctat tgcagcagga 1080
cgggggggag cgcaaacaga tgaaaatcaa gcagcagatg gtgatccaag atatgcattt 1140
ggtagacaac atggtcaaaa aactaccaca acaggagaaa cacctgagag atttacatat 1200
atagcacatc aagatacagg aagatatcca gaaggagatt ggattcaaaa tattaacttt 1260
aaccttcctg taacaaatga taatgtattg ctaccaacag atccaattgg aggtaaagca 1320
ggaattaact atactaatat atttaatact tatggtcctt taactgcatt aaataatgta 1380
ccaccagttt atccaaatgg tcaaatttgg gataaagaat ttgatactga cttaaaacca 1440
agacttcatg taaatgcacc atttgtttgt caaaataatt gtcctggtca attatttgta 1500
aaagttgcgc ctaatttaac aaatgaatat gatcctgatg catctgctaa tatgtcaaga 1560
attgtaactt actcagattt ttggtggaaa ggtaaattag tatttaaagc taaactaaga 1620
gcctctcata cttggaatcc aattcaacaa atgagtatta atgtagataa ccaatttaac 1680
tatgtaccaa gtaatattgg aggtatgaaa attgtatatg aaaaatctca actagcacct 1740
agaaaattat actaa 1755
<210> 6
<211> 1755
<212> DNA
<213> 人工合成
<400> 6
atgagtgatg gagcagttca accagacggt ggtcagcctg ctgtcagaaa tgaaagagct 60
acaggatctg ggaacgggtc tggaggcggg ggtggtggtg gttctggggg tgtggggatt 120
tctacgggta ctttcaataa tcagacggaa tttaaatttt tggaaaacgg atgggtggaa 180
atcacagcaa actcaagcag acttgtacat ttaaatatgc cagaaagtga aaattataga 240
agagtggttg taaataattt ggataaaact gcagttaacg gaaacatggc tttagatgat 300
acccatgcac aaattgtaac accttggtca ttggttgatg caaatgcttg gggagtttgg 360
tttaatccag gagattggca actaattgtt aatactatga gtgagttgca tttagttagt 420
tttgaacaag aaatttttaa tgttgtttta aagactgttt cagaatctgc tactcagcca 480
ccaactaaag tttataataa tgatttaact gcatcattga tggttgcatt agatagtaat 540
aatactatgc catttactcc agcagctatg agatctgaga cattgggttt ttatccatgg 600
aaaccaacca taccaactcc atggagatat tattttcaat gggatagaac attaatacca 660
tctcatactg gaactagtgg cacaccaaca aatatatacc atggtacaga tccagatgat 720
gttcaatttt acactattga aaattctgtg ccagtacact tactaagaac aggtgatgaa 780
tttgctacag gaacatttta ttttgattgt aaaccatgta gactaacaca cacatggcaa 840
acaaatagag cattgggctt accaccattt ctaaattctt tgcctcaagc tgaaggaggt 900
actaactttg gttatatagg agttcaacaa gataaaagac gtggtgtaac tcaaatggga 960
aatacaaaca ttattactga agctactatt atgagaccag ctgaggttgg ttatagtgca 1020
ccatattatt cttttgaggc gtctacacaa gggccattta aaacacctat tgcagcagga 1080
cgggggggag cgcaaacaga tgaaaatcaa gcagcagatg gtgatccaag atatgcattt 1140
ggtagacaac atggtcaaaa aactaccaca acaggagaaa cacctgagag atttacatat 1200
atagcacatc aagatacagg aagatatcca gaaggagatt ggattcaaaa tattaacttt 1260
aaccttcctg taacagatga taatgtattg ctaccaacag atccaattgg aggtaaagca 1320
ggaattaact atactaatat atttaatact tatggtcctt taactgcatt aaataatgta 1380
ccaccagttt atccaaatgg tcaaatttgg gataaagaat ttgatactga cttaaaacca 1440
agacttcatg taaatgcacc atttgtttgt caaaataatt gtcctggtca attatttgta 1500
aaagttgcgc ctaatttaac aaatgaatat gatcctgatg catctgctaa tatgtcaaga 1560
attgtaactt actcagattt ttggtggaaa ggtaaattgg tatttaaagc taaactaaga 1620
gcctctcata cttggaatcc aattcaacaa atgagtatta atgtagataa ccaatttaac 1680
tatgtaccaa gtaatattgg aggtatgaaa attgtatatg aaaaatctca actagcacct 1740
agaaaattat actaa 1755
<210> 7
<211> 1755
<212> DNA
<213> 人工合成
<400> 7
atgagtgatg gaggagttca accagacggt ggtcaacctg ctgtcagaaa tgaaagagct 60
acaggatctg ggaacgggtc tggaggcggg ggtggtggtg gttctggggg tgtggggatt 120
tctacgggta cttttaataa tcagacggaa tttaaatttt tggaaaacgg atgggtggaa 180
atcacagcaa actcaagcag acttgtgcat ttaaatatgc cagaaagtga aaattataga 240
agagtggttg taaataattt ggataaaact gcagttaacg gaaacatggc tttagatgat 300
actcatgcac aaattgtaac accttggtca ttggttgatg caaatgcttg gggagtttgg 360
tttaatccag gagattggca actaattgtt aatactatga gtgagttgca tttagttagt 420
tttgaacaag aaatttttaa tgttgtttta aagactgttt cagaatctgc tactcagcca 480
ccaactaaag tttataataa tgatttaact gcatcattga tggttgcatt agatagtaat 540
aatactatgc catttactcc agcagctatg agatctgaga cattgggttt ttatccatgg 600
aaaccaacca taccaactcc atggagatat tattttcaat gggatagaac attaatacca 660
tctcatactg gaactagtgg cacaccaaca aatatatacc atggtacaga tccagatgat 720
gttcaatttt acactattga aaattctgtg ccagtacact tactaagaac aggtgatgaa 780
tttgctacag gaacatttta ttttgattgt aaaccatgta gactaacaca tacatggcaa 840
acaaatagag cattgggctt accaccattt ctaaattctt tgcctcaagc tgaaggaggt 900
actaactttg gttatatagg agttcaacaa gataaaagac gtggtgtaac tcaaatggga 960
aacacaaaca ttattactga agctactatt atgagaccag ctgaggttgg ttatagtgca 1020
ccatattatt cttttgaggc gtctacacaa gggccattta aaacacctat tgcagcagga 1080
cgggggggag cgcaaacaga tgaaaatcga gcagcagatg gtgatccaag atatgcattt 1140
ggtagacaac atggtcaaaa aactaccaca acaggagaaa cacctgagag atttacatat 1200
atagcacatc aagatacagg aagatatcca gaaggagatt ggattcaaaa tattaacttt 1260
aaccttcctg taacagaaga taatgtattg ctaccaacag atccaattgg aggtaaaaca 1320
ggaattaact atactaatat atttaatact tatggtcctt taactgcatt aaataatgta 1380
ccaccagttt atccaaatgg tcaaatttgg gataaagaat ttgatactga cttaaaacca 1440
agacttcatg taaatgcacc atttgtttgt caaaataatt gtcctggtca attatttgta 1500
aaagttgcac ctaatttaac aaatgaatat gatcctgatg catctgctaa tatgtcaaga 1560
attgtaactt actcagattt ttggtggaaa ggtaaattag tatttaaagc taaactaaga 1620
gcctctcata cttggaatcc aattcaacaa atgagtatca atgtagataa ccaatttaac 1680
tatgtaccaa gtaatattgg aggtatgaaa attgtatatg aaaaatctca actagcacct 1740
agaaaattat attaa 1755
<210> 8
<211> 28
<212> DNA
<213> 人工合成
<400> 8
tatactagta tgagtgacgg tgccgtgc 28
<210> 9
<211> 32
<212> DNA
<213> 人工合成
<400> 9
tatacgcgtt tagtacagct tacgtggagc ga 32
<210> 10
<211> 30
<212> DNA
<213> 人工合成
<400> 10
tatgcggccg catgtctgac ggtgctgtgc 30
<210> 11
<211> 34
<212> DNA
<213> 人工合成
<400> 11
tataagcttt tagtacagct tacgtggagc cagt 34
<210> 12
<211> 28
<212> DNA
<213> 人工合成
<400> 12
tatcccggga tgagcgacgg tggtgtgc 28
<210> 13
<211> 31
<212> DNA
<213> 人工合成
<400> 13
tatctcgagt tagtacagct tacgaggggc c 31
<210> 14
<211> 17
<212> DNA
<213> 人工合成
<400> 14
gttttcccag tcacgac 17
<210> 15
<211> 19
<212> DNA
<213> 人工合成
<400> 15
ccgatcgatc tagcttagc 19
<210> 16
<211> 19
<212> DNA
<213> 人工合成
<400> 16
gtcgctaagc tagatcgat 19
<210> 17
<211> 17
<212> DNA
<213> 人工合成
<400> 17
caggaaacag ctatgac 17
<210> 18
<211> 584
<212> PRT
<213> 人工合成
<400> 18
Met Ser Asp Gly Ala Val Gln Pro Asp Gly Gly Gln Pro Ala Val Arg
1 5 10 15
Asn Glu Arg Ala Thr Gly Ser Gly Asn Gly Ser Gly Gly Gly Gly Gly
20 25 30
Gly Gly Ser Gly Gly Val Gly Ile Ser Thr Gly Thr Phe Asn Asn Gln
35 40 45
Thr Glu Phe Lys Phe Leu Glu Asn Gly Trp Val Glu Ile Thr Ala Asn
50 55 60
Ser Ser Arg Leu Val His Leu Asn Met Pro Glu Ser Glu Asn Tyr Arg
65 70 75 80
Arg Val Val Val Asn Asn Leu Asp Lys Thr Ala Val Asn Gly Asn Met
85 90 95
Ala Leu Asp Asp Thr His Ala Gln Ile Val Thr Pro Trp Ser Leu Val
100 105 110
Asp Ala Asn Ala Trp Gly Val Trp Phe Asn Pro Gly Asp Trp Gln Leu
115 120 125
Ile Val Asn Thr Met Ser Glu Leu His Leu Val Ser Phe Glu Gln Glu
130 135 140
Ile Phe Asn Val Val Leu Lys Thr Val Ser Glu Ser Ala Thr Gln Pro
145 150 155 160
Pro Thr Lys Val Tyr Asn Asn Asp Leu Thr Ala Ser Leu Met Val Ala
165 170 175
Leu Asp Ser Asn Asn Thr Met Pro Phe Thr Pro Ala Ala Met Arg Ser
180 185 190
Glu Thr Leu Gly Phe Tyr Pro Trp Lys Pro Thr Ile Pro Thr Pro Trp
195 200 205
Arg Tyr Tyr Phe Gln Trp Asp Arg Thr Leu Ile Pro Ser His Thr Gly
210 215 220
Thr Ser Gly Thr Pro Thr Asn Ile Tyr His Gly Thr Asp Pro Asp Asp
225 230 235 240
Val Gln Phe Tyr Thr Ile Glu Asn Ser Val Pro Val His Leu Leu Arg
245 250 255
Thr Gly Asp Glu Phe Ala Thr Gly Thr Phe Tyr Phe Asp Cys Lys Pro
260 265 270
Cys Arg Leu Thr His Thr Trp Gln Thr Asn Arg Ala Leu Gly Leu Pro
275 280 285
Pro Phe Leu Asn Ser Leu Pro Gln Ala Glu Gly Gly Thr Asn Phe Gly
290 295 300
Tyr Ile Gly Val Gln Gln Asp Lys Arg Arg Gly Val Thr Gln Met Gly
305 310 315 320
Asn Thr Asn Ile Ile Thr Glu Ala Thr Ile Met Arg Pro Ala Glu Val
325 330 335
Gly Tyr Ser Ala Pro Tyr Tyr Ser Phe Glu Ala Ser Thr Gln Gly Pro
340 345 350
Phe Lys Thr Pro Ile Ala Ala Gly Arg Gly Gly Ala Gln Thr Asp Glu
355 360 365
Asn Gln Ala Ala Asp Gly Asp Pro Arg Tyr Ala Phe Gly Arg Gln His
370 375 380
Gly Gln Lys Thr Thr Thr Thr Gly Glu Thr Pro Glu Arg Phe Thr Tyr
385 390 395 400
Ile Ala His Gln Asp Thr Gly Arg Tyr Pro Glu Gly Asp Trp Ile Gln
405 410 415
Asn Ile Asn Phe Asn Leu Pro Val Thr Asn Asp Asn Val Leu Leu Pro
420 425 430
Thr Asp Pro Ile Gly Gly Lys Ala Gly Ile Asn Tyr Thr Asn Ile Phe
435 440 445
Asn Thr Tyr Gly Pro Leu Thr Ala Leu Asn Asn Val Pro Pro Val Tyr
450 455 460
Pro Asn Gly Gln Ile Trp Asp Lys Glu Phe Asp Thr Asp Leu Lys Pro
465 470 475 480
Arg Leu His Val Asn Ala Pro Phe Val Cys Gln Asn Asn Cys Pro Gly
485 490 495
Gln Leu Phe Val Lys Val Ala Pro Asn Leu Thr Asn Glu Tyr Asp Pro
500 505 510
Asp Ala Ser Ala Asn Met Ser Arg Ile Val Thr Tyr Ser Asp Phe Trp
515 520 525
Trp Lys Gly Lys Leu Val Phe Lys Ala Lys Leu Arg Ala Ser His Thr
530 535 540
Trp Asn Pro Ile Gln Gln Met Ser Ile Asn Val Asp Asn Gln Phe Asn
545 550 555 560
Tyr Val Pro Ser Asn Ile Gly Gly Met Lys Ile Val Tyr Glu Lys Ser
565 570 575
Gln Leu Ala Pro Arg Lys Leu Tyr
580
<210> 19
<211> 584
<212> PRT
<213> 人工合成
<400> 19
Met Ser Asp Gly Ala Val Gln Pro Asp Gly Gly Gln Pro Ala Val Arg
1 5 10 15
Asn Glu Arg Ala Thr Gly Ser Gly Asn Gly Ser Gly Gly Gly Gly Gly
20 25 30
Gly Gly Ser Gly Gly Val Gly Ile Ser Thr Gly Thr Phe Asn Asn Gln
35 40 45
Thr Glu Phe Lys Phe Leu Glu Asn Gly Trp Val Glu Ile Thr Ala Asn
50 55 60
Ser Ser Arg Leu Val His Leu Asn Met Pro Glu Ser Glu Asn Tyr Arg
65 70 75 80
Arg Val Val Val Asn Asn Leu Asp Lys Thr Ala Val Asn Gly Asn Met
85 90 95
Ala Leu Asp Asp Thr His Ala Gln Ile Val Thr Pro Trp Ser Leu Val
100 105 110
Asp Ala Asn Ala Trp Gly Val Trp Phe Asn Pro Gly Asp Trp Gln Leu
115 120 125
Ile Val Asn Thr Met Ser Glu Leu His Leu Val Ser Phe Glu Gln Glu
130 135 140
Ile Phe Asn Val Val Leu Lys Thr Val Ser Glu Ser Ala Thr Gln Pro
145 150 155 160
Pro Thr Lys Val Tyr Asn Asn Asp Leu Thr Ala Ser Leu Met Val Ala
165 170 175
Leu Asp Ser Asn Asn Thr Met Pro Phe Thr Pro Ala Ala Met Arg Ser
180 185 190
Glu Thr Leu Gly Phe Tyr Pro Trp Lys Pro Thr Ile Pro Thr Pro Trp
195 200 205
Arg Tyr Tyr Phe Gln Trp Asp Arg Thr Leu Ile Pro Ser His Thr Gly
210 215 220
Thr Ser Gly Thr Pro Thr Asn Ile Tyr His Gly Thr Asp Pro Asp Asp
225 230 235 240
Val Gln Phe Tyr Thr Ile Glu Asn Ser Val Pro Val His Leu Leu Arg
245 250 255
Thr Gly Asp Glu Phe Ala Thr Gly Thr Phe Tyr Phe Asp Cys Lys Pro
260 265 270
Cys Arg Leu Thr His Thr Trp Gln Thr Asn Arg Ala Leu Gly Leu Pro
275 280 285
Pro Phe Leu Asn Ser Leu Pro Gln Ala Glu Gly Gly Thr Asn Phe Gly
290 295 300
Tyr Ile Gly Val Gln Gln Asp Lys Arg Arg Gly Val Thr Gln Met Gly
305 310 315 320
Asn Thr Asn Ile Ile Thr Glu Ala Thr Ile Met Arg Pro Ala Glu Val
325 330 335
Gly Tyr Ser Ala Pro Tyr Tyr Ser Phe Glu Ala Ser Thr Gln Gly Pro
340 345 350
Phe Lys Thr Pro Ile Ala Ala Gly Arg Gly Gly Ala Gln Thr Asp Glu
355 360 365
Asn Gln Ala Ala Asp Gly Asp Pro Arg Tyr Ala Phe Gly Arg Gln His
370 375 380
Gly Gln Lys Thr Thr Thr Thr Gly Glu Thr Pro Glu Arg Phe Thr Tyr
385 390 395 400
Ile Ala His Gln Asp Thr Gly Arg Tyr Pro Glu Gly Asp Trp Ile Gln
405 410 415
Asn Ile Asn Phe Asn Leu Pro Val Thr Asp Asp Asn Val Leu Leu Pro
420 425 430
Thr Asp Pro Ile Gly Gly Lys Ala Gly Ile Asn Tyr Thr Asn Ile Phe
435 440 445
Asn Thr Tyr Gly Pro Leu Thr Ala Leu Asn Asn Val Pro Pro Val Tyr
450 455 460
Pro Asn Gly Gln Ile Trp Asp Lys Glu Phe Asp Thr Asp Leu Lys Pro
465 470 475 480
Arg Leu His Val Asn Ala Pro Phe Val Cys Gln Asn Asn Cys Pro Gly
485 490 495
Gln Leu Phe Val Lys Val Ala Pro Asn Leu Thr Asn Glu Tyr Asp Pro
500 505 510
Asp Ala Ser Ala Asn Met Ser Arg Ile Val Thr Tyr Ser Asp Phe Trp
515 520 525
Trp Lys Gly Lys Leu Val Phe Lys Ala Lys Leu Arg Ala Ser His Thr
530 535 540
Trp Asn Pro Ile Gln Gln Met Ser Ile Asn Val Asp Asn Gln Phe Asn
545 550 555 560
Tyr Val Pro Ser Asn Ile Gly Gly Met Lys Ile Val Tyr Glu Lys Ser
565 570 575
Gln Leu Ala Pro Arg Lys Leu Tyr
580
<210> 20
<211> 584
<212> PRT
<213> 人工合成
<400> 20
Met Ser Asp Gly Gly Val Gln Pro Asp Gly Gly Gln Pro Ala Val Arg
1 5 10 15
Asn Glu Arg Ala Thr Gly Ser Gly Asn Gly Ser Gly Gly Gly Gly Gly
20 25 30
Gly Gly Ser Gly Gly Val Gly Ile Ser Thr Gly Thr Phe Asn Asn Gln
35 40 45
Thr Glu Phe Lys Phe Leu Glu Asn Gly Trp Val Glu Ile Thr Ala Asn
50 55 60
Ser Ser Arg Leu Val His Leu Asn Met Pro Glu Ser Glu Asn Tyr Arg
65 70 75 80
Arg Val Val Val Asn Asn Leu Asp Lys Thr Ala Val Asn Gly Asn Met
85 90 95
Ala Leu Asp Asp Thr His Ala Gln Ile Val Thr Pro Trp Ser Leu Val
100 105 110
Asp Ala Asn Ala Trp Gly Val Trp Phe Asn Pro Gly Asp Trp Gln Leu
115 120 125
Ile Val Asn Thr Met Ser Glu Leu His Leu Val Ser Phe Glu Gln Glu
130 135 140
Ile Phe Asn Val Val Leu Lys Thr Val Ser Glu Ser Ala Thr Gln Pro
145 150 155 160
Pro Thr Lys Val Tyr Asn Asn Asp Leu Thr Ala Ser Leu Met Val Ala
165 170 175
Leu Asp Ser Asn Asn Thr Met Pro Phe Thr Pro Ala Ala Met Arg Ser
180 185 190
Glu Thr Leu Gly Phe Tyr Pro Trp Lys Pro Thr Ile Pro Thr Pro Trp
195 200 205
Arg Tyr Tyr Phe Gln Trp Asp Arg Thr Leu Ile Pro Ser His Thr Gly
210 215 220
Thr Ser Gly Thr Pro Thr Asn Ile Tyr His Gly Thr Asp Pro Asp Asp
225 230 235 240
Val Gln Phe Tyr Thr Ile Glu Asn Ser Val Pro Val His Leu Leu Arg
245 250 255
Thr Gly Asp Glu Phe Ala Thr Gly Thr Phe Tyr Phe Asp Cys Lys Pro
260 265 270
Cys Arg Leu Thr His Thr Trp Gln Thr Asn Arg Ala Leu Gly Leu Pro
275 280 285
Pro Phe Leu Asn Ser Leu Pro Gln Ala Glu Gly Gly Thr Asn Phe Gly
290 295 300
Tyr Ile Gly Val Gln Gln Asp Lys Arg Arg Gly Val Thr Gln Met Gly
305 310 315 320
Asn Thr Asn Ile Ile Thr Glu Ala Thr Ile Met Arg Pro Ala Glu Val
325 330 335
Gly Tyr Ser Ala Pro Tyr Tyr Ser Phe Glu Ala Ser Thr Gln Gly Pro
340 345 350
Phe Lys Thr Pro Ile Ala Ala Gly Arg Gly Gly Ala Gln Thr Asp Glu
355 360 365
Asn Arg Ala Ala Asp Gly Asp Pro Arg Tyr Ala Phe Gly Arg Gln His
370 375 380
Gly Gln Lys Thr Thr Thr Thr Gly Glu Thr Pro Glu Arg Phe Thr Tyr
385 390 395 400
Ile Ala His Gln Asp Thr Gly Arg Tyr Pro Glu Gly Asp Trp Ile Gln
405 410 415
Asn Ile Asn Phe Asn Leu Pro Val Thr Glu Asp Asn Val Leu Leu Pro
420 425 430
Thr Asp Pro Ile Gly Gly Lys Thr Gly Ile Asn Tyr Thr Asn Ile Phe
435 440 445
Asn Thr Tyr Gly Pro Leu Thr Ala Leu Asn Asn Val Pro Pro Val Tyr
450 455 460
Pro Asn Gly Gln Ile Trp Asp Lys Glu Phe Asp Thr Asp Leu Lys Pro
465 470 475 480
Arg Leu His Val Asn Ala Pro Phe Val Cys Gln Asn Asn Cys Pro Gly
485 490 495
Gln Leu Phe Val Lys Val Ala Pro Asn Leu Thr Asn Glu Tyr Asp Pro
500 505 510
Asp Ala Ser Ala Asn Met Ser Arg Ile Val Thr Tyr Ser Asp Phe Trp
515 520 525
Trp Lys Gly Lys Leu Val Phe Lys Ala Lys Leu Arg Ala Ser His Thr
530 535 540
Trp Asn Pro Ile Gln Gln Met Ser Ile Asn Val Asp Asn Gln Phe Asn
545 550 555 560
Tyr Val Pro Ser Asn Ile Gly Gly Met Lys Ile Val Tyr Glu Lys Ser
565 570 575
Gln Leu Ala Pro Arg Lys Leu Tyr
580
<210> 21
<211> 6163
<212> DNA
<213> 人工合成
<400> 21
ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga 60
ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc 120
attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 180
agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 240
tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 300
ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 360
ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 420
aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 480
ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat 540
attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 600
tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 660
gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 720
tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 780
aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 840
cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 900
agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 960
ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 1020
tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 1080
tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 1140
caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1200
accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1260
attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1320
ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1380
taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1440
taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1500
aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1560
agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1620
ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1680
ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1740
cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1800
tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1860
tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1920
tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1980
tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 2040
ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 2100
acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 2160
ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2220
gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2280
ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2340
ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2400
taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2460
cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2520
tctgtgcggt atttcacacc gcagaccagc cgcgtaacct ggcaaaatcg gttacggttg 2580
agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa 2640
caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga 2700
aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact 2760
cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg 2820
gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc 2880
gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac 2940
ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc 3000
ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag 3060
cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat 3120
ctcactacgc ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc 3180
ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta 3240
atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag 3300
atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat 3360
gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt 3420
gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg 3480
cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga 3540
aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg 3600
agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc 3660
gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg 3720
aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg 3780
cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc 3840
aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag 3900
tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct 3960
atagttctag tggttggcct acgtacccgt agtggctatg gcagggcttg ccgccccgac 4020
gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga 4080
aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg 4140
ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt 4200
ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc 4260
tcccccatct cccctagcac catggctcga gatcccgggt gatcaagtct tcgtcgagtg 4320
attgtaaata aaatgtaatt tacagtatag tattttaatt aatatacaaa tgatttgata 4380
ataattctta tttaactata atatattgtg ttgggttgaa ttaaaggtcc gtatactata 4440
gttctagtgg ttggcctacg tacccgtagt ggctatggca gggcttgccg ccccgacgtt 4500
ggctgcgagc cctgggcctt cacccgaact tgggggttgg ggtggggaaa aggaagaaac 4560
gcgggcgtat tggtcccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga 4620
ccgaaccccg cgtttatgaa caaacgaccc aacacccgtg cgttttattc tgtcttttta 4680
ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc 4740
cccatctccc gtttaaacgg taccgcatgc tatgcatcag ctgctagcag tcttcgtcga 4800
gtgattgtaa ataaaatgta atttacagta tagtatttta attaatatac aaatgatttg 4860
ataataattc ttatttaact ataatatatt gtgttgggtt gaattaaagg tccgtatagt 4920
cgctaagcta gatcgatcgg aatattaata gatcatggag ataattaaaa tgataaccat 4980
ctcgcaaata aataagtatt ttactgtttt cgtaacagtt ttgtaataaa aaaacctata 5040
aatattccgg attattcata ccgtcccacc atcgggcgcg gatcccggtc cgaagcgcgc 5100
ggaattcaaa ggcctacgtc gacgagctca ctagtacgcg tgtcgagaag tactagagga 5160
tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc 5220
tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag 5280
cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5340
cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct 5400
gatcactgct tgagcctagg agatccgtat actccggaat attaatagat catggagata 5460
attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg 5520
taataaaaaa acctataaat attccggatt attcataccg tcccaccatc gggcgcggat 5580
cctagtcgcg gccgctttcg aatctagagc ctgcagtctc gacaagcttg tcgagaagta 5640
ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct 5700
cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt 5760
tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5820
atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 5880
ctggatctga tcactgcttg agcctaggag atccgaacca gataagtgaa atctagttcc 5940
aaactatttt gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc 6000
tattttgtca ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc 6060
caactatttt gtccgcccac agcggggcat ttttcttcct gttatgtttt taatcaaaca 6120
tcctgccaac tccatgtgac aaaccgtcat cttcggctac ttt 6163
Claims (8)
1.一种犬细小病毒的三价抗原，其特征在于，所述三价抗原是由犬细小病毒new CPV-2a的VP2的氨基酸序列、犬细小病毒new CPV-2b的VP2的氨基酸序列和犬细小病毒CPV-2c的VP2的氨基酸序列组成的；所述犬细小病毒new CPV-2a的VP2的氨基酸序列如SEQ ID NO.18所示；所述犬细小病毒new CPV-2b的VP2的氨基酸序列如SEQ ID NO.19所示；所述犬细小病毒CPV-2c的VP2的氨基酸序列如SEQ ID NO.20所示；将犬细小病毒new CPV-2a的VP2的基因序列连接至转移载体PFF的Spe I和Mlu I酶切位点、犬细小病毒new CPV-2b的VP2的基因序列连接至转移载体PFF的Not I 和Hind III酶切位点、犬细小病毒CPV-2c的VP2的基因序列连接至转移载体PFF的Sma I 和Xho I酶切位点；转移载体pFF的序列如SEQ ID NO.21所示。
2.一种重组杆状病毒，其特征在于，所述重组杆状病毒是以E.coli DH10 Bac为出发菌，表达权利要求1所述三价抗原获得的重组微生物，然后利用重组微生物感染Sf9细胞、Sf21细胞或High Five细胞获得的。
3.一种犬细小病毒的三价病毒样颗粒，其特征在于，所述三价病毒样颗粒是利用权利要求2所述的重组杆状病毒感染Sf9细胞、Sf21细胞或High Five细胞，经过离心后收集细胞培养物经过细胞裂解处理后获得的。
4.权利要求所3述的三价病毒样颗粒的制备方法，其特征在于，所述方法如下：
步骤1：将犬细小病毒new CPV-2a的VP2的基因序列、犬细小病毒new CPV-2b的VP2的基因序列和犬细小病毒CPV-2c的VP2的基因序列连接至转移载体pFF获得重组载体pFF-CP-ABC；所述犬细小病毒new CPV-2a的VP2的基因序列如SEQ ID NO.2所示；犬细小病毒newCPV-2b的VP2的基因序列如SEQ ID NO.3所示；犬细小病毒CPV-2c的VP2的基因序列如SEQID NO.4所示；将犬细小病毒new CPV-2a的VP2的基因序列连接至转移载体PFF的Spe I和Mlu I酶切位点、犬细小病毒new CPV-2b的VP2的基因序列连接至转移载体PFF的Not I 和Hind III酶切位点、犬细小病毒CPV-2c的VP2的基因序列连接至转移载体PFF的Sma I 和Xho I酶切位点；转移载体pFF的序列如SEQ ID NO.21所示；
步骤2：将重组载体pFF-CP-ABC利用热休克方法转化E.coli DH10 Bac感受态获得重组杆状病毒CPV-ABC；
步骤3：将重组杆状病毒CPV-ABC转染贴壁Sf9细胞，收集细胞培养物，离心后分离上清液和细胞，向细胞加入NaHCO3悬浮处理后，离心后获取上清液。
5.根据权利要求4所述的制备方法，其特征在于，步骤1中获得转移载体pFF的方法如下：
步骤1.1：pFastBacDual载体p10启动子下的MCS中，将Kpn I至Nhe I的序列删除；将pH启动子下MCS中的Rsr II至Spe I的序列删除获得质粒pFD(k+n)(r+s)；
步骤1.2：将SEQ ID NO.1所示的10H的基因序列，通过BstZ17 I酶切位点连入质粒pFD(k+n)(r+s)获得转移载体pFF，转移载体pFF的序列如SEQ ID NO.21所示。
6.一种犬细小病毒的三价疫苗，其特征在于，所述三价疫苗由权利要求3所述的三价病毒样颗粒和Montanide Gel 02 PR佐剂组成。
7.权利要求1所述的三价抗原或权利要求6所述的三价疫苗在制备预防或治疗犬细小病毒性肠炎的疫苗或药物中的应用。
8.权利要求1所述的三价抗原或权利要求3所述的三价病毒样颗粒在制备犬细小病毒性肠炎的三价卵黄抗体中的应用。
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012089748A1 (en) * | 2010-12-29 | 2012-07-05 | Intervet International B.V. | Canine babesiosis vaccine antigen |
| CN102791288A (zh) * | 2010-03-05 | 2012-11-21 | 英特维特国际股份有限公司 | 重组的减毒细小病毒 |
| CN108273055A (zh) * | 2018-01-03 | 2018-07-13 | 复旦大学 | 犬细小病毒疫苗组合物及其制备方法与应用 |
| CN108348594A (zh) * | 2015-09-29 | 2018-07-31 | 梅里亚股份有限公司 | 犬科动物细小病毒(cpv)病毒样颗粒(vlp)疫苗及其用途 |
| CN110157687A (zh) * | 2019-05-25 | 2019-08-23 | 青岛易邦生物工程有限公司 | 一种犬细小病毒三价亚单位疫苗 |
-
2022
- 2022-05-26 CN CN202210584627.0A patent/CN114957406B/zh active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102791288A (zh) * | 2010-03-05 | 2012-11-21 | 英特维特国际股份有限公司 | 重组的减毒细小病毒 |
| WO2012089748A1 (en) * | 2010-12-29 | 2012-07-05 | Intervet International B.V. | Canine babesiosis vaccine antigen |
| CN108348594A (zh) * | 2015-09-29 | 2018-07-31 | 梅里亚股份有限公司 | 犬科动物细小病毒(cpv)病毒样颗粒(vlp)疫苗及其用途 |
| CN108273055A (zh) * | 2018-01-03 | 2018-07-13 | 复旦大学 | 犬细小病毒疫苗组合物及其制备方法与应用 |
| CN110157687A (zh) * | 2019-05-25 | 2019-08-23 | 青岛易邦生物工程有限公司 | 一种犬细小病毒三价亚单位疫苗 |
Non-Patent Citations (6)
| Title |
|---|
| "Baculovirus-Produced Influenza Virus-like Particles in Mammalian Cells Protect Mice from Lethal Influenza Challenge";Xian-Chun Tang 等;《Viral Immunology》;第24卷(第4期);摘要 * |
| "HPV-16-11 L1双价重组杆状病毒的构建及VLP形成";庄敏 等;《中华微生物学和免疫学杂志》(第9期);第771-776页 * |
| "Recombinant canine parvovirus-like particles express foreign epitopes in silkworm pupae";Hao Feng 等;《Vet Microbiol》;第154卷(第1-2期);第49-57页 * |
| Liu,D.F.等."VP2 [Canine parvovirus 2b]".《genbank》.2012,ACCESSION NO.AFK94137. * |
| Zhao,H.等."capsid protein VP2 [Canine parvovirus]".《genbank》.2015,ACCESSION NO.AKN46016. * |
| Zhao,H.等."capsid protein VP2 [Canine parvovirus]".《genbank》.2015,ACCESSION NO.AKN46049. * |
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