CN114292823A - 携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株及其构建方法和应用 - Google Patents
携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株及其构建方法和应用 Download PDFInfo
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Abstract
本发明属于动物基因工程疫苗领域,具体涉及携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株及其构建方法和应用。所述重组LaSota疫苗株是用基因VII型NDV的HN基因和碱性蛋白酶裂解位点附近氨基酸序列突变的F基因分别替换LaSota疫苗株的HN和F基因后得到的。具体的,该重组LaSota疫苗株是在宿主菌中利用同源重组技术和反向筛选标记系统制备得到。该重组LaSota疫苗株为研制防控基因VII型NDV感染的新型、高效单联或多联的弱毒活苗和灭活疫苗提供制苗毒株和合适载体。
Description
技术领域
本发明属于动物基因工程疫苗领域,具体涉及携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株及其构建方法和应用。
背景技术
新城疫(Newcastle disease,ND)是由新城疫病毒(Newcastle disease virus,NDV)感染引起的能够感染各种禽类的急性、高度接触性和致死性传染病,严重危害我国和世界养禽业的健康发展。NDV基因组编码6个结构蛋白,即核衣壳蛋白(NP)、磷蛋白(P)、基质蛋白(M)、融合蛋白(F)、血凝素-神经氨酸酶(HN)和大聚合酶(L)。其中,F和HN 是NDV的两种功能性囊膜表面糖蛋白。F蛋白介导病毒囊膜与细胞膜融合,且F蛋白中的蛋白酶裂解位点与NDV的毒力密切相关。HN将病毒吸附到宿主细胞受体上,触发F蛋白将病毒与宿主细胞膜的融合,从而使病毒核衣壳复合物进入宿主细胞。HN蛋白的神经氨酸酶活性可以降解唾液酸受体,从而避免子代病毒的自我吸附和聚集,使得子代病毒能顺利脱离宿主细胞。NDV虽然只有一个血清型,但根据F基因的核苷酸序列同源性,NDV 可被划分为I和II两类共18个基因型,目前我国NDV的流行株以基因VII型为主。
疫苗免疫是防控传染病的重要手段。现有用于防控ND的商品化疫苗主要包括单联弱毒活苗和多联灭活疫苗。与单苗相比较,多联疫苗在控制病原的混合感染和降低疫苗接种劳动强度、减少动物应激等方面具有显著优势。传统的多联疫苗生产需要分别制备多种抗原,生产工艺繁琐,生产成本高。利用合适载体制备的基因工程多联疫苗能够克服传统多联疫苗制备工艺的缺陷。NDVLaSota株作为防控ND的弱毒活疫苗株,在防控ND的发生和流行过程中发挥了重要作用。NDVLaSota株具有良好的安全性和在鸡胚中高滴度复制等优点。LaSota株属于基因II型NDV,除了作为疫苗株之外,还可以作为基因工程疫苗的载体。国内外利用属于基因II型的LaSota疫苗株作为载体,采用传统的酶切和连接技术构建了表达多种外源基因的基因工程多联疫苗。
但由于我国鸡群中普遍存在高水平抗NDV LaSota株的母源抗体,使得基于LaSota株的重组活载体疫苗的临床应用受到很大限制。此外,我国ND的流行发生了很大变化,目前我国流行的NDV毒株主要是基因VII型。而LaSota株和目前流行的VII型NDV毒株在F和HN基因上存在很多差异,从而导致基于LaSota株的疫苗不能有效控制我国目前流行的基因VII型NDV的感染。虽然我国科研工作者已经利用基于VII型NDV的反向遗传技术,通过突变F基因的碱性蛋白酶裂解位点附近的氨基酸,使基因VII型NDV强毒株的毒力减弱,制备出用于制备灭活疫苗用的VII型NDV种毒,并获得了防控基因VII型NDV 的单联灭活疫苗新兽药证书,但由于新兽药证书的限制和该疫苗毒株是基于VII型NDV流行株,只是F蛋白的碱性蛋白酶裂解位点附近的几个氨基酸发生了替换,作为弱毒活疫苗使用,存在毒力返强的安全性的问题等限制。
申请公布号为CN102533676A的中国发明专利申请,公开了表达基因VII型F和HN重组新城疫弱毒疫苗A-NDV-LX/I4,其构建方法为:将中国主要流行的NDV基因VII型分离株JS-5-05-Go的囊膜糖蛋白基因F基因的强毒裂解位点突变为弱毒位点,再将JS-5-05-Go 的致弱的囊膜糖蛋白基因F基因和HN基因片段与NDV/LX株基因组的对应部分进行替换,拯救构建出致弱的表达基因VII型F和HN重组新城疫弱毒疫苗A-NDV-LX/I4。不过其在拯救病毒的过程中,是将pCI-NP、pCI-P和pCI-L3个真核表达质粒与含有NDV基因组 cDNA全长的转录载体(pA-NDV-LX/I4)共转染BSR-T7/5细胞,即其需要用4个质粒共转染细胞,只有当4个质粒同时进入一个细胞时才能拯救出病毒,拯救病毒的效率低。而且在基因替换过程中采用的是传统的基因酶切和连接方法,不但效率低,而且还在病毒基因组中引入额外的非病毒序列,由此导致未知的重组病毒生物学性状的改变和潜在危害。
发明内容
本发明提供了一种含有基因VII型的表达融合蛋白(Fusion,F)和血凝素-神经氨酸酶 (Hemagglutinin-neuraminidase,HN)的基因的嵌合型LaSota弱毒株—rLaSota-7F/HN。本发明以LaSota疫苗株为骨架,在宿主菌大肠杆菌中利用同源重组技术和高效的反向筛选标记系统,将LaSota疫苗株的F和HN基因分别替换为基因VII型NDV流行株的HN基因和碱性蛋白酶裂解位点附近氨基酸突变为NDV弱毒株序列的基因VII型F基因,构建了带有基因VII型F和HN基因的嵌合型LaSota弱毒株——rLaSota-7F/HN,为研制防控基因 VII型NDV感染的新型、高效单联或多联的弱毒活苗和灭活疫苗提供制苗毒株和合适载体。
本发明嵌合型LaSota弱毒株—rLaSota-7F/HN的构建采用以下技术方案:
(1)LaSota的感染性克隆质粒的构建。
具体的,利用重叠PCR将涵盖整个基因组的7个cDNA片段克隆到转录载体pOLTV5中,获得重组质粒pLaSota。
(2)分别构建含有VII型NDV流行毒株的F和HN基因的重组载体。
具体的,分别合成基因VII型NDV流行毒株全长的F和HN基因,并分别克隆在pUC57载体中,得到载体pUC-7F和pUC-7HN;在合成F基因过程中,将基因VII型NDV流行株的F基因中的碱性蛋白酶裂解位点氨基酸序列从112RRQKR↓F117突变为 112GRQGR↓L117。NDV的F基因与毒力相关,而且碱性蛋白酶裂解位点附近的氨基酸序列发挥决定作用,突变这些氨基酸是防止重组病毒的毒力增强。
(3)利用带有LaSota株的F和HN基因两侧同源序列的引物,以步骤(2)中制备的重组载体pUC-7F和pUC-7HN为模板,分别扩增优化突变后的基因VII型NDV的F基因和HN基因,以含有抗性筛选标记amp以及大肠杆菌自杀基因ccdB的重组质粒 p15A-ccdB-amp为模板,扩增得到氨苄青霉素抗性筛选标记(amp)和大肠杆菌自杀基因 (ccdB)表达盒amp-ccdB。
(4)将带有LaSota株F基因两侧同源臂的amp-ccdB表达盒与LaSota的感染性克隆质粒pLaSota共电转化进宿主菌,利用氨苄青霉素抗性筛选F基因被amp-ccdB替换的阳性克隆,得到重组质粒pLaSota-ΔF-amp-ccdB。
(5)利用Redαβ重组酶介导的同源重组技术,将pLaSota-ΔF-amp-ccdB中的amp-ccdB 替换为优化突变后的基因VII型NDV的F基因,得到重组质粒pLaSota-7F。
(6)利用带有LaSota株HN基因两侧同源臂的引物扩增的amp-ccdB表达盒片段与重组质粒pLaSota-7F共电转化进宿主菌,利用氨苄青霉素抗性筛选HN基因被amp-ccdB替换的筛选阳性克隆,得到重组质粒pLaSota-7F-ΔHN-amp-ccdB。
(7)用Redαβ重组酶介导的同源重组技术,将重组质粒pLaSota-7F-ΔHN-amp-ccdB中的amp-ccdB替换为基因VII型NDV的HN基因,得到重组质粒pLaSota-7F-HN。
(8)将重组质粒pLaSota-7F-HN与辅助质粒pCI-NP-P-L共转染BHK-21细胞,收集转染后的细胞裂解液,接种10日龄SPF鸡胚,接种72小时后收获血凝活性阳性的鸡胚尿囊液即为重组病毒rLaSota-7F-HN。
本发明的有益效果为:
本发明采用同源重组的方法构建含有基因VII型F和HN基因的嵌合型LaSota弱毒株,该方法快捷高效,而且不引入任何非病毒基因序列。
本发明在病毒拯救时,重组质粒pLaSota-7F-HN与辅助质粒pCI-NP-P-L共转染BHK-21 细胞,相比现有技术中需要将3个辅助质粒和1个重组质粒共同转染细胞相比,2个质粒被共同转染进一个细胞的几率远远高于4个质粒被共同转染进一个细胞的几率,本发明拯救病毒的效率更高。
附图说明
图1为重组质粒pLaSota的结构示意图。
图2为重组质粒pLaSota-7F的ApaLI酶切鉴定结果图。
图3为重组质粒pLaSota-7F-7HN的图谱。
图4为重组质粒pLaSota-7F-7HN的ApaL I酶切鉴定结果图。
图5为重组质粒pCI-NP-P-L的图谱。
图6为重组病毒rLaSota-7F/HN在SPF鸡胚中的复制动态图。
图7为血凝抑制试验(HI)检测rLaSota-7F/HN疫苗免疫后的抗体水平图。
图8为rLaSota-7F/HN活疫苗对基因VII型NDV攻毒的保护情况。
图9为rLaSota-7F/HN活疫苗免疫后基因VII型NDV攻毒的组织显微病变情况。
图10为rLaSota-7F/HN活疫苗免疫后基因VII型NDV攻毒的病毒载量图。
保藏信息:
鸡新城疫病毒基因VII型中国分离株FJSW2021:
保藏时间:2021年12月13日;
保藏单位名称:中国微生物菌种保藏管理委员会普通微生物中心;
保藏编号:CGMCC NO.45060;
保藏单位地址:北京市朝阳区北辰西路1号院3号;
分类命名:基因VII型新城疫病毒。
重组病毒rLaSota-7F/HN:
保藏时间:2021年12月13日;
保藏单位名称:中国微生物菌种保藏管理委员会普通微生物中心;
保藏编号:CGMCC NO.45059;
保藏单位地址:北京市朝阳区北辰西路1号院3号;
分类命名:携带基因VII型F和HN基因重组Lasota毒株。
具体实施方式
下面通过具体实施方式对本发明进行更加详细的说明,以便于对本发明技术方案的理解,但并不用于对本发明保护范围的限制。
实施例中所用pOLTV5、pUC57、GBred-gyrA462感受态细胞、GB05-dir感受态细胞、top10感受态细胞、BHK-21细胞均为市售产品。
实施例1带有基因VII型NDV流行株HN基因和碱性蛋白酶裂解位点氨基酸突变的F基因的LaSota株感染性克隆的构建
1.1NDV LaSota疫苗株感染性克隆质粒的构建
根据GenBank中公布的NDV LaSota株序列(GenBank登录号AF077761),利用重叠PCR将涵盖整个基因组的7个cDNA片段克隆到转录载体pOLTV5中,获得重组质粒 pLaSota,重组质粒pLaSota如图1所示;需要说明的是:完整的LaSota基因组cDNA上游为T7RNA聚合酶启动子,下游带有δ肝炎病毒(HDV)的核酶序列(ribozyme)和T7转录终止序列。
1.2构建含有VII型NDV流行毒株的F和HN基因的重组载体
根据GenBank数据库中公布的基因VII型NDV流行毒株的基因组序列,分别优化合成全长的F和HN基因,并分别克隆在pUC57载体中,得到载体pUC-7F和pUC-7HN;在合成F基因过程中,将基因VII型NDV流行株的F基因编码的碱性蛋白酶裂解位点氨基酸序列从112RRQKR↓F117突变为112GRQGR↓L117;优化突变后的F基因的序列如SEQ ID NO:3所示,优化合成的HN基因的序列如SEQ ID NO:4所示。优化突变后的F基因编码的蛋白序列见SEQ IDNO:1,优化合成的HN基因编码的蛋白质序列见SEQ ID NO:2。
1.3基因VII型NDV的F基因和HN基因、抗性筛选标记的扩增
利用带有LaSota株的F和HN基因两侧同源序列的引物(见表1),以pUC-7F和 pUC-7HN为模板,分别扩增优化突变后的基因VII型NDV的F基因和优化后的HN基因,以p15A-ccdB-amp质粒(ccdB-amp是插在p15A的NdeI和EcoRI位点之间)为模板,扩增氨苄青霉素抗性筛选标记(amp)和大肠杆菌自杀基因(ccdB)表达盒amp-ccdB。
表1扩增基因VII型NDV毒株F和HN基因以及相应amp-ccdB表达盒的引物
1.4转化筛选重组质粒pLaSota-ΔF-ampccdB
将带有LaSota株的F基因两侧同源臂的amp-ccdB表达盒与LaSota的感染性克隆质粒 pLaSota共电转化进宿主菌GBred-gyrA462感受态细胞,在含有氨苄青霉素的LB琼脂平板上筛选阳性克隆,得到LaSota株的F基因被amp-ccdB替换的重组质粒pLaSota-Δ F-amp-ccdB。
1.5重组质粒pLaSota-7F的构建
使用PacI限制性内切酶37℃过夜酶切pLaSota-ΔF-amp-ccdB质粒,通过T4 DNA聚合酶将线性化的pLaSota-ΔF-amp-ccdB载体与优化突变后的基因VII型NDV的F基因片段进行聚合反应。聚合体系为200ng的优化突变后的基因VII型NDV的F基因片段、2μg 的线性化pLaSota-ΔF-amp-ccdB载体、2μL的10×NEB Buffer 2.1、0.2μL的T4 DNA聚合酶、加入双蒸水补足体系至20μL,反应程序为25℃1h、75℃20min、50℃30min;将反应体系电转化至10%L-阿拉伯糖诱导的GB05-dir感受态细胞中,复苏1h后涂布至带有氯霉素抗性的LB平板,37℃过夜培养;从平板上挑取单菌落扩大培养,提取质粒经ApaLI 限制性内切酶酶切和测序得到重组质粒pLaSota-7F,重组质粒的ApaLI酶切鉴定结果见图 2。
1.6重组质粒pLaSota-7F-ΔHN-amp-ccdB的构建
将利用带有HN基因两侧同源臂的引物扩增的amp-ccdB表达盒片段与重组质粒pLaSota-7F共电转化进宿主菌GBred-gyrA462感受态细胞,在含有氨苄青霉素的LB琼脂平板上筛选阳性克隆,得到LaSota株的HN基因被amp-ccdB替换的重组质粒pLaSota-7F- ΔHN-amp-ccdB。
1.7重组质粒pLaSota-7F-7HN的构建
使用PacI限制性内切酶37℃过夜酶切pLaSota-7F-ΔHN-amp-ccdB质粒,通过T4DNA 聚合酶将线性化的pLaSota-7F-ΔHN-amp-ccdB载体与优化合成的基因VII型NDV的HN基因片段进行聚合反应。聚合体系为200ng的优化合成的基因VII型NDV的HN基因片段、2μg的线性化pLaSota-7F-ΔHN-amp-ccdB载体、2μL的10×NEB Buffer 2.1、0.2μL 的T4 DNA聚合酶、加入双蒸水补足体系至20μL,反应程序为25℃1h、75℃20min、 50℃30min;将反应体系电转化至10%L-阿拉伯糖诱导的GB05-dir感受态细胞中,复苏1 h后涂布至带有氯霉素抗性的LB平板,37℃过夜培养;从平板上挑取单菌落扩大培养,提取质粒经ApaLI限制性内切酶酶切和测序得到重组质粒pLaSota-7F-7HN,质粒图谱见图 3,重组质粒的ApaLI酶切鉴定结果见图4。
实施例2带有基因VII型NDV流行株HN基因和碱性蛋白酶裂解位点氨基酸突变的F基因的重组LaSota株的拯救
2.1表达NDV LaSota疫苗株核蛋白(NP)、磷酸蛋白(P)和大聚合酶蛋白(L)的转录辅助质粒的构建
将编码NDV LaSota疫苗株的大聚合酶蛋白(L)的cDNA序列克隆到pCI-neo真核表达载体的CMV启动子下游;将编码核蛋白(NP)和磷酸蛋白(P)的cDNA通过2A肽序列串联后克隆到pCI-neo真核表达载体的SV40启动子下游,获得同时表达NP、P和L蛋白的转录质粒pCI-NP-P-L。具体采用以下方案:
a、NP-2A-P的制备
以pLaSota质粒为模板,采用针对NP的引物:
NP-F:5’-ATGTCTTCCGTATTTGATGAG-3’;
NP-R:5’-ACGTCACCGCATGTTAGAAGACTTCCTCTGCCCTCATACCCCCAGTCGG-3’;
针对P基因的引物:
P-F:5’-TAACATGCGGTGACGTGGAGGAGAATCCCGGCCCTATGGCCACCTTTAC-3’;
P-R:5’-TTAGCCATTTAGAGCAAGGCGC-3’;分别进行PCR扩增NP和P基因,分别以回收后的NP基因及P基因为模板,以NP-F/P-R引物进行融合PCR的扩增,并回收片段 NP-2A-P。
b、重组质粒pCI-NP-2A-P的构建
以pCI-neo载体为模板,以带有NP基因和P基因同源臂序列的上下游引物TY-pCIneo-F: 5’-CATCAAATACGGAAGACATGGTGGCTCTAGCCTTAAGTTCG-3’和TY-pCIneo-R: 5’-GCCTTGCTCTAAATGGCTAAGCGGGACTCTGGGGTTCGAAATG-3’进行扩增,将回收的扩增片段pCI-neo载体与片段NP-2A-P使用NEBuilder重组试剂盒进行聚合重组,产物转化top10感受态细胞,涂布氨苄抗性平板进行筛选阳性克隆,最终获得重组质粒 pCI-NP-2A-P。
c、重组质粒pCI-NP-P-L的构建
以L-XbaI-F:5’-ACGCGTTCTAGAAAGGCAAAACAGCTC-3’和L-NotI-R:5’-GAATTC GCGGCCGCCCGGGTCGACAATTGGCCAGAAAAG-3’为引物扩增L基因,将纯化的L基因和质粒pCI-NP-2A-P分别进行XbaI+NotI酶切,纯化后使用T4连接酶进行连接,转化top10 感受态细胞,涂布氨苄抗性平板进行筛选阳性克隆,最终得到重组质粒pCI-NP-P-L。重组质粒pCI-NP-P-L的质粒图谱见图5。
2.2重组病毒的拯救
a、转录质粒和辅助质粒转染BHK-21细胞
将BHK-21细胞培养于6孔细胞培养板内生长至80%单层时,用表达T7 RNA聚合酶的重组痘苗病毒vTF7-3以MOI=1感染细胞1h,吸弃重组痘苗病毒vTF7-3感染物,然后将2μg转录质粒pLaSota-7F-7HN与2μg辅助质粒pCI-NP-P-L按照Lipofectamine 3000转染试剂盒操作说明共转染BHK-21细胞。转染后24h,弃去转染混合物,用PBS洗涤细胞 2次,加入含有2%胎牛血清的MEM培养基继续孵育至96h。
b、重组病毒rLaSota-7F/HN的获得
收获培养物上清,接种10日龄的SPF鸡胚并培养120h,收获HA阳性尿囊液即为拯救的重组病毒rLaSota-7F/HN。
实施例3重组病毒rLaSota-7F/HN的生物学特性鉴定
3.1rLaSota-7F/HN的RT-PCR鉴定
利用病毒基因组RNA提取试剂盒分别提取LaSota株感染尿囊液和rLaSota-7F/HN感染鸡胚尿囊液的总RNA。采用针对F基因两侧基因序列设计鉴定引物7F-F: 5′-GCTGCGTCTC TGAGATTGCG-3′和7F-R:5′-GGCCTCTCTTACCGTTCTAC-3′;以及 HN基因两侧的引物7HN-F:5′-GTAGAACGGTAAGAGAGGCC-3′;和7HN-R: 5′-AGCACTGGCTGATTGTGGTC-3′进行RT-PCR;对PCR产物进行序列测定,鉴定显示F 和HN基因的正确插入。
3.2rLaSota-7F/HN的生物学特性分析
按照OIE标准测定第30代rLaSota-7F/-HN的鸡胚平均致死时间(MDT)、鸡胚半数感染量(EID50)、1日龄雏鸡脑内接种致病指数(ICPI)及6周龄鸡静脉接种致病指数(IVPI) 致病性指标。结果显示rLaSota-7F/HN的MDT为130小时,ICPI和IVPI指数均为0,符合弱毒株的标准。接种SPF鸡胚后96小时所收获尿囊液中重组病毒的EID50为108.5/0.1mL。
3.3rLaSota-7F/HN在鸡胚中的生长特性分析
将NDV LaSota疫苗株和第30代rLaSota-7F/HN按每胚102EID50的剂量分别接种10日龄SPF鸡胚,在接种后24h、48h、72h、96h和120h分别收获鸡胚尿囊液,测定EID50,分析LaSota株与rLaSota-7F/HN在鸡胚中的生长特性差异。结果显示,重组病毒 rLaSota-7F/HN与NDVLaSota疫苗株具有相似的在鸡胚中高复制特性,如图6所示。
实施例4重组病毒rLaSota-7F/HN的免疫效力评价
4.1重组rLaSota-7F/HN活疫苗接种
将购自北京梅里亚的60只1周龄白来航SPF鸡随机平均分成3组,各组鸡均分别饲养于负压隔离器中。组1通过点眼滴鼻途径接种100微升含107EID50(半数鸡胚感染剂量) 的重组rLaSota-7F/HN;组2通过点眼滴鼻途径接种100微升的正常鸡胚尿囊液;组3作为不接种的空白对照。疫苗免疫后每周采取各组鸡的血清样品,利用血凝抑制试验(Hemagglutinin inhibition,HI)测定疫苗免疫产生的抗鸡新城疫病毒抗体水平。在免疫后4 周,将组1和组2的鸡分别用105ELD50(半数鸡胚致死剂量)的鸡新城疫病毒基因VII型中国分离株FJSW2021通过点眼滴鼻途径进行攻毒,组3不攻毒作为对照。攻毒后每天观察各组鸡的临床表现并记录鸡的发病率和死亡率,连续观察7天。
在攻毒后每天采集鸡的口咽棉拭子和泄殖腔棉拭子,利用PCR测定攻毒鸡的排毒情况。同时,采集攻毒后发病死亡鸡的肺、气管、腺胃、十二指肠、盲肠扁桃体和法氏囊。将每个器官样品平均分成两份,一份利用荧光定量PCR测定组织中的NDV病毒载量,另一份用于制备组织病理切片,观察组织显微病变情况。
4.2血凝抑制试验(HI)检测疫苗免疫后的抗体水平
疫苗免疫后每周采取静脉血,分离血清后利用HI试验测定疫苗免疫后的抗NDV抗体水平。血凝抑制试验按照常规方法操作:将25微升血清样品进行2倍倍比稀释后转移到V型底的96孔血凝板中,每孔加入25微升的4个血凝单位的基因VII型NDV抗原,震荡混匀后室温静置30分钟,然后每孔加入50微升0.5%鸡红血球,轻微振荡混匀后,室温静置 30分钟观察血凝抑制效果。以能够完全抑制血球凝集的最大血清稀释度的倒数作为抗体的 HI抗体滴度。
4.3实时荧光定量PCR
利用基因VII型新城疫病毒的HN基因作为检测标记物设计引物,进行实时荧光定量 PCR检测组织中的病毒载量。
上、下游引物分别为HN-qF:5’-GCAGAGACCACTCACACTCACA-3’,HN-qR: 5’-TGCAGGACTTCCGATTTTGGGTG-3’。实时荧光定量PCR的反应体系包括10μL of 2x ChamQUniversal SYBR qPCR Master Mix(诺唯赞,南京),上下游引物(10μM)各1μL,1μL cDNA模板,用无核酸酶的纯水补足体积20μL。反应条件为95℃预变性30秒,然后进行95℃10秒和60℃15秒扩增40个循环。将扩增的129-bp NDV HN基因片段克隆入 pMD18-T载体(宝生物工程(大连)有限公司)制备标准质粒。利用109~103拷贝/微升的 10倍倍比稀释的标准质粒借助CFX Maestro软件建立标准曲线。病毒载量计算为每mg组织中的HN基因拷贝数结果报告为三次重复试验的平均值加上标准差。
4.4攻毒鸡的口咽和泄殖腔排毒检测
采用常规RT-PCR检测攻毒鸡的口咽棉拭子和泄殖腔棉拭子中病毒核酸,以便确定排毒情况。RT-PCR引物采用针对基因VII型NDV的特异性引物NDV-F: 5’-GGAAGATCAAACGCCTTGC-3和NDV-R:5’-GACAATCGGGAATGCTAACAGG-3’,扩增产物的大小为325bp。
4.5重组病毒rLaSota-7F/HN活疫苗免疫原性和免疫效力
利用重组病毒作为活疫苗通过点眼滴鼻途径免疫后,采集血清样品进行HI试验,检测疫苗产出的抗基因VII型NDV的抗体水平。检测结果显示,单剂量点眼滴鼻免疫即可诱导鸡体产生高水平的HI抗体,最高抗体滴度达到28(图7)。
在免疫后4周,利用基因VII型NDV强毒分离株FJSW2021进行攻毒,未接种疫苗的鸡表现精神沉郁,羽毛凌乱,在攻毒后第2天表现呼吸困难,张口喘气。病鸡排出绿色水样粪便,且在攻毒后84h开始出现死亡。图8结果显示,rLaSota-7F/HN活疫苗接种组对基因VII型NDV强毒分离株FJSW2021的攻毒提供完全保护,而未免疫攻毒组在攻毒后第4天全部发病死亡。对病死鸡进行剖检发现,未免疫攻毒组所有病死鸡均有新城疫的典型剖检病变,包括腺胃坏死和出血,脾脏肿大。而rLaSota-7F/HN活疫苗接种组和空白对照组的鸡仍然表现临床健康状况,没有出现明显的肉眼病变。死鸡的组织学病变包括肺间质慢性炎症浸润;脾、盲肠扁桃体和法氏囊淋巴细胞坏死;肠上皮和腺胃上皮细胞坏死,而 rLaSota-7F/HN活疫苗免疫组和空白对照组的鸡没有出现上述组织学体征(图9)。
对攻毒后各组鸡的肺脏、气管、脾脏、十二指肠、腺胃、盲肠扁桃体和法氏囊等器官组织中的基因VII型病毒载量利用实时荧光定量PCR进行检测发现,rLaSota-7F/HN活疫苗免疫组和空白对照组的病毒载量没有明显差异,但显著低于未免疫攻毒组(图10)。
为了确定rLaSota-7F/HN活疫苗免疫能否阻止免疫鸡排毒,在攻毒后的第1至第7天分别采集鸡的口咽棉拭子和泄殖腔棉拭子,利用实时荧光定量PCR检测排毒情况。表1结果显示,rLaSota-7F/HN活疫苗免疫组鸡只在免疫后第3天即停止排毒,而未免疫攻毒组在攻毒后持续排毒直至死亡。
表1攻毒后不同天数的排毒情况
以上结果证明,利用本发明构建的重组病毒作为活疫苗免疫,可以为鸡提供针对基因 VII型新城疫病毒感染的良好保护。
以上所述之实施例,只是本发明的较佳实施例而已,并非限制本发明的实施范围,故凡依本发明专利范围所述的构造、特征及原理所做的等效变化或修饰,均应包括于本发明申请专利范围内。
SEQUENCE LISTING
<110> 河南农业大学
<120> 携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株及其构建方法和应
用
<130> 无
<160> 4
<170> PatentIn version 3.3
<210> 1
<211> 540
<212> PRT
<213> 人工序列
<400> 1
Met Leu Ile Thr Arg Ile Met Leu Ile Leu Gly Cys Ile Arg Pro Thr
1 5 10 15
Ser Ser Leu Asp Gly Arg Pro Leu Ala Ala Ala Gly Ile Val Val Thr
20 25 30
Gly Asp Lys Ala Val Asn Val Tyr Thr Ser Ser Gln Thr Gly Ser Ile
35 40 45
Ile Val Lys Leu Leu Pro Asn Met Pro Arg Asp Lys Glu Ala Cys Ala
50 55 60
Lys Ala Pro Leu Glu Ala Tyr Asn Arg Thr Leu Thr Thr Leu Leu Thr
65 70 75 80
Pro Leu Gly Asp Ser Ile Arg Lys Ile Gln Gly Ser Val Ser Thr Ser
85 90 95
Gly Gly Gly Arg Gln Gly Arg Leu Ile Gly Ala Val Ile Gly Ser Val
100 105 110
Ala Leu Gly Val Ala Thr Ala Ala Gln Ile Thr Ala Ala Ala Ala Leu
115 120 125
Ile Gln Ala Asn Arg Asn Ala Ala Asn Ile Leu Arg Leu Lys Glu Ser
130 135 140
Ile Ala Ala Thr Asn Glu Ala Val His Glu Val Thr Asp Gly Leu Ser
145 150 155 160
Gln Leu Ser Val Ala Val Gly Lys Met Gln Gln Phe Val Asn Asp Gln
165 170 175
Phe Asn Asn Thr Ala Arg Glu Leu Asp Cys Ile Lys Ile Thr Gln Gln
180 185 190
Val Gly Val Glu Leu Asn Leu Tyr Leu Thr Glu Leu Thr Thr Val Phe
195 200 205
Gly Pro Gln Ile Thr Ser Pro Ala Leu Thr Gln Leu Thr Ile Gln Ala
210 215 220
Leu Tyr Asn Leu Ala Gly Gly Asn Met Asp His Leu Leu Thr Arg Leu
225 230 235 240
Gly Ile Gly Asn Asn Gln Leu Ser Ser Leu Ile Gly Ser Gly Leu Ile
245 250 255
Thr Gly Tyr Pro Ile Leu Tyr Asp Ser Gln Thr Gln Leu Leu Gly Ile
260 265 270
Gln Val Asn Leu Pro Ser Val Gly Asn Leu Asn Asn Met Arg Ala Thr
275 280 285
Tyr Leu Glu Thr Leu Ser Val Ser Thr Thr Lys Gly Tyr Ala Ser Ala
290 295 300
Leu Val Pro Lys Val Val Thr Gln Val Gly Ser Val Ile Glu Glu Leu
305 310 315 320
Asp Thr Ser Tyr Cys Ile Glu Ser Asp Leu Asp Leu Tyr Cys Thr Arg
325 330 335
Ile Val Thr Leu Pro Met Ser Pro Gly Ile Tyr Ser Cys Leu Ser Gly
340 345 350
Asn Thr Ser Ala Cys Met Tyr Ser Lys Thr Glu Gly Ala Leu Thr Thr
355 360 365
Pro Tyr Met Ala Leu Lys Gly Ser Val Ile Ala Asn Cys Lys Ile Thr
370 375 380
Thr Cys Arg Cys Thr Asp Pro Pro Gly Ile Ile Ser Gln Asn Tyr Gly
385 390 395 400
Glu Ala Val Ser Leu Ile Asp Arg His Leu Cys Asn Val Leu Ser Leu
405 410 415
Asp Gly Ile Thr Leu Arg Leu Ser Gly Glu Phe Asp Ala Thr Tyr Gln
420 425 430
Lys Asn Ile Ser Ile Leu Asp Ser Gln Val Ile Val Thr Gly Asn Leu
435 440 445
Asp Ile Ser Thr Glu Leu Gly Asn Val Asn Asn Ser Ile Ser Asn Ala
450 455 460
Leu Asp Arg Leu Ala Glu Ser Asn Ser Lys Leu Glu Lys Val Asn Val
465 470 475 480
Arg Leu Thr Ser Thr Ser Ala Leu Ile Thr Tyr Ile Val Leu Thr Val
485 490 495
Ile Ser Leu Val Phe Gly Ala Leu Ser Leu Gly Leu Ala Cys Tyr Leu
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<210> 2
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<212> PRT
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Met Asp Arg Ala Val Asn Arg Val Val Leu Glu Asn Glu Glu Arg Glu
1 5 10 15
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20 25 30
Met Val Met Thr Leu Ala Ile Ser Ala Ala Ala Leu Ala Tyr Ser Thr
35 40 45
Gly Ala Ser Thr Pro His Asp Leu Ala Gly Ile Ser Thr Val Ile Ser
50 55 60
Lys Thr Glu Asp Lys Val Thr Ser Leu Leu Ser Leu Ser Gln Asp Val
65 70 75 80
Ile Asp Lys Ile Tyr Lys Gln Val Ala Leu Glu Ser Pro Leu Ala Leu
85 90 95
Leu Asn Thr Glu Ser Ile Ile Met Asn Ala Ile Thr Ser Leu Ser Tyr
100 105 110
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115 120 125
Asp Pro Asp Tyr Ile Gly Gly Ile Gly Lys Glu Leu Ile Val Asp Asp
130 135 140
Ile Ser Asp Val Thr Ser Phe Tyr Pro Ser Ala Tyr Gln Glu His Leu
145 150 155 160
Asn Phe Ile Pro Ala Pro Thr Thr Gly Ser Gly Cys Thr Arg Ile Pro
165 170 175
Ser Phe Asp Met Ser Thr Thr His Tyr Cys Tyr Thr His Asn Val Ile
180 185 190
Leu Ser Gly Cys Arg Asp His Ser His Ser His Gln Tyr Leu Ala Leu
195 200 205
Gly Val Leu Arg Thr Ser Ala Thr Gly Arg Val Phe Phe Ser Thr Leu
210 215 220
Arg Ser Thr Asn Leu Asp Asp Thr Gln Asn Arg Lys Ser Cys Ser Val
225 230 235 240
Ser Ala Thr Pro Leu Gly Cys Asp Met Leu Cys Ser Lys Val Thr Glu
245 250 255
Thr Glu Glu Glu Asp Tyr Lys Ser Val Ala Pro Thr Ser Met Val His
260 265 270
Gly Arg Leu Gly Phe Asp Gly Gln Tyr His Glu Lys Asp Leu Asp Thr
275 280 285
Thr Val Leu Phe Lys Asp Trp Val Ala Asn Tyr Pro Gly Ala Gly Gly
290 295 300
Gly Ser Phe Ile Asp Asp Arg Val Trp Phe Pro Val Tyr Gly Gly Leu
305 310 315 320
Lys Pro Asn Ser Pro Ser Asp Thr Ala Gln Glu Gly Lys Tyr Val Ile
325 330 335
Tyr Lys Arg His Asn Asn Thr Cys Pro Asp Glu Gln Asp Tyr Gln Ile
340 345 350
Arg Met Ala Lys Ser Ser Tyr Lys Pro Gly Arg Phe Gly Gly Lys Arg
355 360 365
Val Gln Gln Ala Ile Leu Ser Ile Lys Val Ser Thr Ser Leu Gly Lys
370 375 380
Asp Pro Val Leu Thr Ile Pro Pro Asn Thr Ile Thr Leu Met Gly Ala
385 390 395 400
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405 410 415
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465 470 475 480
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Ser Thr Cys Phe Lys Val Val Lys Thr Asn Lys Thr Tyr Cys Leu Ser
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<210> 3
<211> 1623
<212> DNA
<213> 人工序列
<400> 3
atgcttatca cacggatcat gttgatattg ggatgcatcc gcccaacttc ttctctcgat 60
ggtcgcccgc tcgcagcagc agggatagta gttaccggtg acaaagcagt aaacgtatat 120
acttcttccc agacgggtag catcatcgtt aaacttctcc ccaacatgcc gcgggacaag 180
gaggcatgtg ccaaagcccc cctcgaagcg tataaccgca cactcacaac actcctcacg 240
cctttgggag atagcatacg caaaattcag gggtccgttt caactagcgg gggaggacgg 300
caagggaggc tcataggggc tgtgataggt tccgtcgctc ttggagtagc cactgcggcc 360
caaatcacgg cagccgctgc acttatacag gcaaacagaa atgccgccaa catactgcgc 420
ctcaaagagt caatagccgc gacgaacgaa gcggtgcacg aggtgactga cggtctctct 480
cagttgagcg tcgccgtggg aaaaatgcag caattcgtca acgaccagtt taacaatacg 540
gcacgcgaac tcgactgcat aaaaattact cagcaagttg gggttgaact caatttgtat 600
ttgaccgagc ttaccactgt attcggaccc caaattacct ccccggctct tacgcaattg 660
acaattcagg ccctgtacaa tttggcaggt gggaatatgg accacttgct cactaggctg 720
gggattggaa acaatcaact ctcctcactg atagggtccg gtttgatcac tggctatccg 780
atactgtacg actctcaaac ccagctgctc ggtatccaag ttaaccttcc cagtgtggga 840
aatctcaata atatgcgggc aacttacctg gaaacacttt ccgtctctac cactaaaggc 900
tatgcgagcg ctctggttcc caaagtagtt acgcaagtgg gaagcgtcat cgaagaattg 960
gatacatcct attgcattga gagcgatttg gacttgtact gcacgcggat tgtcacactg 1020
ccgatgtcac caggaatcta ctcatgcctt tccggcaata ccagtgcctg tatgtactct 1080
aagacggagg gagccctcac taccccatac atggcgttga aaggtagcgt tatagcaaac 1140
tgcaaaataa caacatgtcg ctgtaccgac ccaccaggaa taataagtca aaactacggt 1200
gaagccgtga gcctgatcga cagacacctt tgcaatgtct tgagcttgga cgggatcact 1260
cttcgcctgt caggggaatt tgacgcgacg taccagaaga atatatccat tcttgatagc 1320
caagtcattg tgacagggaa tttggacatc agtaccgagc ttggcaatgt gaacaactct 1380
atttcaaacg ctcttgatag gttggcggaa agcaattcca agttggagaa agtaaacgtg 1440
cgccttacat caacgagcgc gctcataacg tacatcgttc ttacagtcat aagcctcgtc 1500
ttcggagcgt tgagcctggg tctcgcttgt tatctgatgt acaaacaaaa ggcgcaacaa 1560
aagacgcttc tgtggcttgg caacaataca ctcgaccaaa tgcgcgctac cactagggcg 1620
tga 1623
<210> 4
<211> 1716
<212> DNA
<213> 人工序列
<400> 4
atggatagag ctgtaaatag agttgtgctt gaaaatgagg agagagaggc aaaaaataca 60
tggagactcg tctttcgcat agcggtactc ttgcttatgg taatgaccct tgcgatttct 120
gcggcagccc ttgcatattc aacgggagca agcacaccgc acgaccttgc ggggatttcc 180
acagtcatat caaagacgga agataaagtg acttcactcc tgtcactcag tcaggacgta 240
atagacaaga tctacaagca agtcgccttg gagagcccac tggctctcct taacactgaa 300
agtattatta tgaacgctat cacgtcactc agttaccaaa taaacggagc cgctaacaac 360
agtgggtgcg gtgccccagt tcacgacccc gattatatag gcggtattgg aaaggagctg 420
atagtggacg acatttcaga tgttactagt ttctatccgt ctgcgtacca ggagcacttg 480
aatttcatac ctgccccaac tacggggagt ggttgtacca ggatccctag cttcgatatg 540
agcacgactc attattgtta tacacacaat gtcatccttt caggatgtag agaccatagc 600
cactctcatc aatatctcgc tttgggagtg ctgcgcacca gtgctacggg aagagtattc 660
tttagcactc tgaggagtac caatctggat gacacccaaa acagaaaaag ctgtagcgtc 720
tctgcgaccc cgttggggtg tgacatgctc tgtagcaagg tcacagagac ggaagaggaa 780
gattataaat ccgttgcgcc gacctcaatg gtacatggtc gcttgggttt cgacggacaa 840
taccacgaga aagacctcga tacaacggtg ttgttcaagg attgggttgc taattaccct 900
ggtgcgggag gagggagttt cattgacgat cgggtgtggt ttccggtgta cggcgggctg 960
aaaccaaact ctccctctga tactgcacag gaaggaaaat atgtaatcta taagcggcat 1020
aacaacactt gtccagacga acaagattat cagatcagaa tggcaaagtc aagttacaag 1080
cctgggcggt ttggggggaa gcgcgtacaa caggccatct tgagtattaa agttagcact 1140
tcattgggca aagacccagt gttgaccatt cctccgaaca ctatcacgct catgggtgca 1200
gagggacgga tactcaccgt gggaacctcc catttccttt atcaacgggg aagcagctac 1260
ttcagtcctg cactgctcta cccaatgacc gtgaataata agacagctac tctccattct 1320
ccctatacat ttaatgcttt cactagacca ggatccgtac catgccaggc atccgcgcgg 1380
tgccctaaca gctgcatcac aggagtgtac accgacccat atccacttat ttttcatcgc 1440
aaccatacac ttcgcggggt tttcggcacc atgctcgatg acgagcaggc aaggttgaac 1500
cccgtttctg cggtgtttga caacatctct cggagtaggg tcactcgcgt cagctcttct 1560
tccacgaagg cggcatatac aacgtccacg tgcttcaagg ttgtcaaaac caacaaaacc 1620
tattgcctgt ctattgcgga aatcagcaat acgttgtttg gtgagttcag aattgtccca 1680
ctgctggtag agatcttgaa agacgaccgg gtctaa 1716
Claims (10)
1.携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株,其特征在于,所述重组LaSota疫苗株是用基因VII型NDV的HN基因和碱性蛋白酶裂解位点附近氨基酸序列突变的F基因分别替换LaSota疫苗株的HN和F基因后得到的。
2.根据权利要求1所述的携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株,其特征在于,所述重组LaSota疫苗株是以表达LaSota全基因组的载体为骨架,在大肠杆菌中利用同源重组技术和反向筛选标记系统,将LaSota疫苗株的F和HN基因分别替换为基因VII型NDV流行株的HN基因和碱性蛋白酶裂解位点附近氨基酸优化突变后的基因VII型F基因,得到重组的转录质粒;然后将转录质粒和表达NDV LaSota疫苗株核蛋白、磷酸蛋白和大聚合酶蛋白的辅助质粒共转染细胞进行病毒拯救,进而得到重组LaSota疫苗株;优化突变后的基因VII型F基因编码蛋白的氨基酸序列如SEQ ID NO:1所示。
3.根据权利要求1所述的携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株,其特征在于,所述基因VII型NDV流行株的HN基因编码的蛋白的氨基酸序列如SEQ ID NO:2所示。
4.携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株的构建方法,其特征在于,包括以下步骤:
(1)利用pOLTV5构建LaSota株的感染性克隆质粒pLaSota;
(2)分别构建含有VII型NDV流行毒株的HN基因和优化突变后的F基因的重组载体;其中基因VII型NDV流行株的F基因中的碱性蛋白酶裂解位点氨基酸序列从112RRQKR↓F117突变为112GRQGR↓L117;
(3)利用带有LaSota株的F和HN基因两侧同源臂的引物,以步骤(2)中制备的重组载体、含有抗性筛选标记以及大肠杆菌自杀基因的重组质粒为模板,分别扩增优化突变后的基因VII型NDV的F基因和HN基因,以及抗性筛选标记和宿主菌自杀基因的筛选表达盒;
(4)将带有LaSota株F基因两侧同源臂的筛选表达盒与LaSota株的感染性克隆质粒共转化进宿主菌,筛选阳性克隆,得到重组质粒pLaSota-ΔF-筛选表达盒;
(5)利用Redαβ重组酶介导的同源重组技术,将pLaSota-ΔF-筛选表达盒中的筛选表达盒替换为优化突变后的基因VII型NDV的F基因,得到重组质粒pLaSota-7F;
(6)利用带有LaSota株HN基因两侧同源臂的筛选表达盒与重组质粒pLaSota-7F共转化进宿主菌,筛选阳性克隆,得到重组质粒pLaSota-7F-ΔHN-筛选表达盒;
(7)用Redαβ重组酶介导的同源重组技术,将重组质粒pLaSota-7F-ΔHN-筛选表达盒中的筛选表达盒替换为基因VII型NDV的HN基因,得到重组质粒pLaSota-7F-HN;
(8)将重组质粒pLaSota-7F-HN与表达NDV LaSota疫苗株核蛋白、磷酸蛋白和大聚合酶蛋白的辅助质粒共转染目标细胞,收集转染后的细胞裂解液,接种鸡胚,收获血凝活性阳性的鸡胚尿囊液,即得到携带基因VII型新城疫病毒F和HN基因的重组LaSota疫苗株rLaSota-7F/HN。
5.根据权利要求4所述的构建方法,其特征在于,步骤(1)中,利用重叠PCR将涵盖整个基因组的cDNA片段克隆到转录载体pOLTV5中,获得重组质粒pLaSota。
6.根据权利要求4所述的构建方法,其特征在于,所述抗性筛选标记为青霉素抗性筛选标记,大肠杆菌自杀基因为大肠杆菌自杀基因ccdB。
7.根据权利要求4所述的构建方法,其特征在于,所述辅助质粒的构建方法为:
将编码NDV LaSota疫苗株的大聚合酶蛋白的cDNA序列克隆到pCI-neo真核表达载体的CMV启动子下游;将编码核蛋白、磷酸蛋白的cDNA通过2A肽序列串联后克隆到pCI-neo真核表达载体的SV40启动子下游,获得同时表达核蛋白、磷酸蛋白和大聚合酶蛋白的辅助质粒。
8.权利要求1所述重组LaSota疫苗株在制备预防基因VII型NDV所致疫病的疫苗中的应用。
9.权利要求1所述重组LaSota疫苗株在制备基因VII型NDV诊断试剂和治疗药物中的应用。
10.权利要求1所述重组LaSota疫苗株作为基因工程载体的应用。
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---|---|---|---|---|
CN111575247A (zh) * | 2020-05-22 | 2020-08-25 | 浙江鼎持生物制品有限公司 | 一种新城疫嵌合病毒标记疫苗株及其构建方法和应用 |
WO2021103421A1 (zh) * | 2019-11-27 | 2021-06-03 | 青岛易邦生物工程有限公司 | 基因vii型新城疫病毒弱毒株及其应用 |
CN113736800A (zh) * | 2021-10-08 | 2021-12-03 | 中国动物卫生与流行病学中心 | 一种鸽源新城疫病毒重组疫苗株及其构建方法和应用 |
-
2021
- 2021-12-17 CN CN202111550232.0A patent/CN114292823A/zh not_active Withdrawn
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2021103421A1 (zh) * | 2019-11-27 | 2021-06-03 | 青岛易邦生物工程有限公司 | 基因vii型新城疫病毒弱毒株及其应用 |
CN111575247A (zh) * | 2020-05-22 | 2020-08-25 | 浙江鼎持生物制品有限公司 | 一种新城疫嵌合病毒标记疫苗株及其构建方法和应用 |
CN113736800A (zh) * | 2021-10-08 | 2021-12-03 | 中国动物卫生与流行病学中心 | 一种鸽源新城疫病毒重组疫苗株及其构建方法和应用 |
Non-Patent Citations (3)
Title |
---|
LIU,Y.L. ET AL.: "fusion protein [Avian orthoavulavirus 1],GenBank: AAL18935.3", 《GENBANK》, pages 1 - 2 * |
LIU,Y.L. ET AL.: "hemagglutinin-neuraminidase [Avian orthoavulavirus 1],GenBank: AAL18936.2", 《GENBANK》, pages 1 - 2 * |
QILONG QIAO ET AL.: "Single-Dose Vaccination of Recombinant Chimeric Newcastle Disease Virus (NDV) LaSota Vaccine Strain Expressing Infectious Bursal Disease Virus (IBDV) VP2 Gene Provides Full Protection against Genotype VII NDV and IBDV Challenge", 《VACCINES》, vol. 9, pages 1 - 17 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117757891A (zh) * | 2024-02-22 | 2024-03-26 | 潍坊华卓生物科技有限公司 | 预防h9亚型禽流感病毒的功能益生菌反向筛选方法及应用 |
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