重组动物疱疹病毒活载体疫苗的构建与应用研究进展

doi:10.16431/j.cnki.1671-7236.2022.01.036

摘要/Abstract

摘要： 重组病毒活载体疫苗是通过基因工程方法构建的活载体疫苗，规避了传统疫苗的一些缺点，具有极大的优势和应用前景。目前被广泛用作重组病毒载体的动物疱疹病毒有火鸡疱疹病毒（Herpesvirus of turkey，HVT）、伪狂犬病病毒（Pseudorabies virus，PRV）、鸭瘟病毒（Duck enteritis virus，DEV）、鸡传染性喉气管炎病毒（Infectious laryngotracheitis virus，ILTV）等。对疱疹病毒基因组序列进行分析显示，其基因组大，复制非必需区域多，容纳外源基因的能力强。同时疱疹病毒具有宿主范围广泛、免疫持续期长、安全性较好及相关基因操作技术成熟等优点。笔者总结了当前构建重组动物疱疹病毒的主要方法，包括传统同源重组技术、细菌人工染色体（BAC）技术、Fosmid文库及CRISPR/Cas9基因编辑技术，简述了各个方法的基本原理及技术特点，比较了各个方法的优缺点，并分析了各个方法的最适运用条件，从而更准确地为研究工作提供理论及技术参考；对外源基因的表达方式进行分析，论证了不同启动子对外源基因表达的调控作用不同，同时对不同疱疹病毒的常用插入位点进行了研究成果的列举总结，并对当前重组病毒HVT、PRV、DEV及ILTV活载体疫苗的研究进展进行总结，对相关生物制品的注册应用信息进行了归纳，介绍了高选择率外源基因及高成功率插入位点，从而为相关生物制品的研制提供理论参考。

关键词: 重组动物疱疹病毒; 活载体疫苗; 火鸡疱疹病毒(HVT); 伪狂犬病病毒(PRV); 鸭瘟病毒(DEV); 传染性喉气管炎病毒(ILTV)

Abstract: Recombinant virus live vector vaccines are live vector vaccines constructed through genetic engineering methods, avoiding some of the shortcomings of traditional vaccines, and have great advantages and application prospects. Animal Herpesviruses currently widely used as recombinant virus vectors include Herpesvirus of turkey (HVT), Pseudorabies virus (PRV), Duck enteritis virus (DEV), and avian Infectious laryngotracheitis virus (ILTV), etc. Analysis of the Herpesvirus genome sequence shows that its genome is large, there are many unnecessary regions for replication, and the ability to accommodate exogenous gene is strong. At the same time, the Herpesvirus has the advantages of wide host range, long duration of immunity, good safety and mature related gene manipulation technology. This article summarized the main methods of constructing recombinant animal Herpesviruses, including traditional homologous recombination technology, bacterial artificial chromosome technology (BAC), Fosmid library and CRISPR/Cas9 gene editing technology, and briefly described the basic principles and technical characteristics of each method, compared the advantages and disadvantages of each method, analyzed the optimal application conditions of each method, so as to provide more accurate theoretical and technical references for research work. The expression of exogenous genes were analyzed, and examples were given to demonstrate the different regulatory effects of different promoters on the expression of exogenous genes. At the same time, the common insertion sites of different Herpesviruses were listed and summarized. The research progress and the registration application information of related biological products of the current recombinant viruses HVT, PRV, DEV and ILTV live vector vaccines were summarized, and the high-selectivity exogenous genes and high-success rate insertion sites were induced, so as to provide a theoretical reference for the development of related biological products.

Key words: recombinant animal Herpesvirus; live vector vaccine; Herpesvirus of turkey (HVT); Pseudorabies virus (PRV); Duck enteritis virus (DEV); Infectious laryngotracheitis virus (ILTV)

中图分类号:

S852.65⁺9.1

杨明珠, 陈晓春, 宋佳诚, 侯亚欣, 李俊平. 重组动物疱疹病毒活载体疫苗的构建与应用研究进展[J]. 中国畜牧兽医, 2022, 49(1): 338-351.

YANG Mingzhu, CHEN Xiaochun, SONG Jiacheng, HOU Yaxin, LI Junping. Research Progress on Construction and Application of Recombinant Animal Herpesvirus Live Vector Vaccine[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 338-351.

参考文献

[1] 刘乃芝,亓秀晔,郭杨丽,等.基因工程重组乳酸菌防控动物疫病的研究进展[J].中国畜牧兽医,2020,47(4):1199-1208. LIU N Z,QI X Y,GUO Y L,et al.Research progress of genetic engineering recombinant lactic acid bacteria on epidemic prevention and control of animal diseases[J].China Animal Husbandry & Veterinary Medicine,2020,47(4):1199-1208.(in Chinese)
[2] 胡林玲,郭松林,关瑞章,等.爱德华氏菌疫苗的研究进展[J].安徽农业科学,2013,41(25):10353-10355. HU L L,GUO S L,GUAN R Z,et al.Research advance of the vaccine against Edwardsiella spp.[J].Journal of Anhui Agricultural Sciences,2013,41(25):10353-10355.(in Chinese)
[3] 高映雪,薛瑞,林敏,等.表达猪圆环病毒2型的多拷贝P28-Cap重组猪痘病毒的构建及表达量分析[J].中国畜牧兽医,2021,48(3):818-828. GAO Y X,XUE R,LIN M,et al.Construction and expression analysis of multicopies P28-Cap recombinant Swine pox virus expressing Porcine circovirus type 2[J].China Animal Husbandry & Veterinary Medicine,2021,48(3):818-828.(in Chinese)
[4] 程坚,刘秀梵,彭大新,等.表达鸡Ⅱ型干扰素基因的重组鸡痘病毒的构建[J].农业生物技术学报,2002,2:152-155. CHENG J,LIU X F,PENG D X,et al.Construction of recombinant Fowlpox virus expressing chicken type Ⅱ interferon[J].Journal of Agricultural Biotechnology,2002,2:152-155.(in Chinese)
[5] 许晨旭.共表达H5亚型AIV HA基因和鸡IL-6基因重组鸡痘病毒的构建及免疫效力评价[D].扬州:扬州大学,2014. XU C X.Construction and immune efficacy of recombinant Fowlpox virus co-expressing HA gene of H5 subtype AIV and chicken IL-6 genes[D].Yangzhou:Yangzhou University,2014.(in Chinese)
[6] KIM D H,PARK B J,AHN H S,et al.Canine interferon lambda 3 expressed using an adenoviral vector effectively induces antiviral activity against Canine influenza virus[J].Virus Research,2021,296:198342.
[7] 陈伟伟,崔海燕,温晔,等.水痘带状疱疹病毒重组腺病毒载体疫苗的构建及评价[J].中国生物制品学杂志,2019,32(1):1-6. CHEN W W,CUI H Y,WEN Y,et al.Construction and evaluation of recombinant Adenovirus expressing glycoprotein E gene against Varicella zoster virus[J].Chinese Journal of Biologicals,2019,32(1):1-6.(in Chinese)
[8] 王梦阁,吴英,程安春,等.禽疱疹病毒活载体疫苗研究进展[J].中国农业科技导报,2020,22(3):85-93. WANG M G,WU Y,CHENG A C,et al.Research progress on live vector vaccine of Avian herpes virus[J].Journal of Agricultural Science and Technology,2020,22(3):85-93.(in Chinese)
[9] ERTL H C.Viral vectors as vaccine carriers[J].Current Opinion in Virology,2016,21:1-8.
[10] 赵军,黄剑波,朱玲,等.伪狂犬病毒表达载体的研究进展[J].猪业科学,2016,33(2):102-104. ZHAO J,HUANG J B,ZHU L,et al.Research progress of Pseudorabies virus expression vector[J].Swine Industry Science,2016,33(2):102-104.(in Chinese)
[11] 赵银龙,李健,朱海霞,等.同源重组介导的重组病毒研究进展[J].江苏农业科学,2016,44(11):1-6. ZHAO Y L,LI J,ZHU H X,et al.Research progress of recombinant viruses mediated by homologous recombination[J].Jiangsu Agricultural Sciences,2016,44(11):1-6.(in Chinese)
[12] 刘召明,张丽萍,王秀丽,等.重组马立克病毒通用转移载体的构建及初步应用[J].中国动物检疫,2008,25(1):26-28. LIU Z M,ZHANG L P,WANG X L,et al.Construction and preliminary application of the universal transferring vector of Marek's disease virus expressing GFP gene[J].China Animal Health Inspection,2008,25(1):26-28.(in Chinese)
[13] 时庆贺,马宁宁,刘占,等.共表达PCV2 Cap蛋白和IL-2重组变异株伪狂犬病病毒构建及对小鼠的免疫原性[J].中国兽医学报,2021,41(3):480-486. SHI Q H,MA N N,LIU Z,et al.Construction of recombinant mutated Pseudorabies virus co-expressing PCV2 Cap protein and IL-2 and its immunogenicity in mice[J].Chinese Journal of Veterinary Science,2021,41(3):480-486.(in Chinese)
[14] ANDOH K,YAMAZAKI K,HONDA Y,et al.Turkey herpesvirus with an insertion in the UL3-4 region displays an appropriate balance between growth activity and antibody-eliciting capacity and is suitable for the establishment of a recombinant vaccine[J].Archives of Virology,2017,162(4):931-941.
[15] 张华伟,周明光,侯真真,等.基于CRISPR/Cas9介导的同源重组技术构建TK、gE和gI基因缺失的伪狂犬病病毒[J].华中农业大学学报,2021,40(2):206-212. ZHANG H W,ZHOU M G,HOU Z Z,et al.Construction of Pseudorabies virus with TK,gE and gI gene deletions based on CRISPR/Cas9-mediated homologous recombination techniques[J].Journal of Huazhong Agricultural University,2021,40(2):206-212.(in Chinese)
[16] 兰德松,石星明,王云峰,等.利用Red/ET技术构建表达H5亚型禽流感病毒HA基因的重组火鸡疱疹病毒[J].微生物学报,2009,49(1):78-84. LAN D S,SHI X M,WAN Y F,et al.Construction of a recombinant HVT virus expressing the HA gene of Avian influenza virus H5N1via Red/ET recombination system[J].Acta Microbiologica Sinica,2009,49(1):78-84.(in Chinese)
[17] ARNUAD R,ANDREI P,DEVIKA N,et al.ESI mutagenesis:A one-step method for introducing mutations into bacterial artificial chromosomes[J].Life Science Alliance,2021,4(2):e202000836.
[18] 陈柳,倪征,余斌,等.重组鸭瘟病毒载体中筛选高效表达鸭坦布苏病毒E蛋白启动子[J].中国农业科学,2020,53(24):5125-5134. CHEN L,NI Z,YU B,et al.Optimized promoter regulating of Duck Tembusu virus E protein expression delivered by a vectored Duck enteritis virus in vitro[J].Scientia Agricultura Sinica,2020,53(24):5125-5134.(in Chinese)
[19] 朱佳.表达IBDV VP2蛋白的重组火鸡疱疹病毒疫苗的构建及其免疫效力试验[D].扬州:扬州大学,2015. ZHU J.Construction of a recombinant Herpesvirus vaccine strain of turkey expressing IBDV VP2 protein and the its protection efficacy[D].Yangzhou:Yangzhou University,2015.(in Chinese)
[20] 刘娅梅,乔永峰,顾一奇,等.猪源伪狂犬病病毒变异株AH02LA株细菌人工染色体的构建与鉴定[J].中国预防兽医学报,2018,40(10):880-885. LIU Y M,QIAO Y F,GU Y Q,et al.Construction and characterization of bacterial artificial chromosome of a Pseudorabies virus strain AH02LA isolated from a dead piglet[J].Chinese Journal of Preventive Veterinary Medicine,2018,40(10):880-885.(in Chinese)
[21] 王志胜,刘芳,许梦微,等.一种新型火鸡疱疹病毒细菌人工染色体的构建与鉴定[J].畜牧兽医学报,2016,47(10):2071-2080. WANG Z S,LIU F,XU M W,et al.Construction and identification of a novel bacterial artificial chromosome of Herpesvirus of turkey[J].Acta Veterinaria et Zootechnica Sinica,2016,47(10):2071-2080.(in Chinese)
[22] 李奇蒙,陈普成,柳金雄,等.表达传染性法氏囊病毒VP2蛋白的重组火鸡疱疹病毒的构建[J].中国预防兽医学报,2014,36(7):515-518. LI Q M,CHEN P C,LIU J X,et al.Construction of recombinant Herpesvirus of turkeys expressing the VP2 protein of Infectious bursal disease virus[J].Chinese Journal of Preventive Veterinary Medicine,2014,36(7):515-518.(in Chinese)
[23] 吴红霞.伪狂犬病病毒变异株和经典株神经传导和毒力差异的研究[D].北京:中国农业科学院,2021. WU H X.Differences in nerve conduction and virulence between variant and classical Pseudorabies virus strains[D].Beijing:Chinese Academy of Agricultural Sciences,2021.(in Chinese)
[24] ABID M,TEKLUE T,LI Y F,et al.Generation and immunogenicity of a recombinant Pseudorabies virus co-expressing Classical swine fever virus E2 protein and Porcine circovirus type 2 capsid protein based on Fosmid Library Platform[J].Pathogens,2019,8(4):279.
[25] 徐长江,王晓朋,徐奎,等.利用CRISPR/Cas9编辑系统构建pAPN基因敲除的IPI-2I细胞系[J].中国畜牧兽医,2021,48(7):2282-2290. XU C J,WANG X P,XU K,et al.Estabilshment of pAPN gene knockout IPI-2I cell lines mediated by CRISPR/Cas9 system[J].China Animal Husbandry & Veterinary Medicine,2021,48(7):2282-2290.(in Chinese)
[26] 檀克勤,马现永,崔艺燕,等.利用CRISPR/Cas9和λ-Red级联技术构建产肠毒素大肠杆菌LT敲除菌株[J].中国畜牧兽医,2020,47(3):666-675. TAN K Q,MA X Y,CUI Y Y,et al.Construction of enterotoxigenic E.coli LT knockout strain using CRISPR/Cas9 and λ-Red cascaded technology[J].China Animal Husbandry & Veterinary Medicine,2020,47(3):666-675.(in Chinese)
[27] 白银荣.利用CRISPR/Cas9技术构建表达EGFP的两株重组火鸡疱疹病毒rHVT-US2/US10-EGFP[D].呼和浩特:内蒙古农业大学,2018. BAI Y R.Construction of two recombinant Herpesviruses of turkey expressing EGFP rHVT-US2/US10-EGFP using CRISPR/Cas9 technology[D].Hohhot:Inner Mongolia Agricultural University,2018.(in Chinese)
[28] CHANG P X,AMEEN F,SEALY J E,et al.Application of HDR-CRISPR/Cas9 and erythrocyte binding for rapid generation of recombinant Turkey herpesvirus-vectored Avian influenza virus vaccines[J].Vaccines,2019,7(4):192.
[29] 吴凤笋,李文刚,樊淑华.应用伪狂犬病毒表达载体研制重组疫苗的研究进展[J].郑州牧业工程高等专科学校学报,2006,26(3):21-23. WU F S,LI W G,FAN S H.Research progress of using Pseudorabies virus expression vector to develop recombinant vaccine[J].Journal of Zhengzhou College of Animal Husbandry Engineering,2006,26(3):21-23.(in Chinese)
[30] 彭芸芸,仲晓宁,李春蕾.伪狂犬病毒载体的研究进展[J].山东畜牧兽医,2013,34(5):75-76. PENG Y Y,ZHONG X N,LI C L,et al.Research progress of Pseudorabies virus vector[J].Shandong Journal of Animal Science and Veterinary Medicine,2013,34(5):75-76.(in Chinese)
[31] 杜文娟.表达猪圆环病毒2型(PCV2)Cap蛋白重组伪狂犬病病毒的构建及其生物学特性鉴定[D].北京:中国农业科学院,2016. DU W J.Construction and biological characterization of a recombinant Pseudorabies virus expressing capsid protein of Porcine circovirus type 2[D].Beijing:Chinese Academy of Agricultural Sciences,2016.(in Chinese)
[32] 王子书,王萍,许健,等.共表达鸡传染性法氏囊病病毒LX株VP2基因及鸡新城疫病毒PX02/3株F基因的重组火鸡疱疹病毒的构建[J].中国兽医杂志,2020,56(11):26-30. WANG Z S,WANG P,XU J,et al.Construction of recombinant Turkey herpesvirus co-expressing Infectious bursal disease virus LX strain VP2 gene and chicken Newcastle disease virus PX02/3 strain F gene[J].Chinese Journal of Veterinary Medicine,2020,56(11):26-30.(in Chinese)
[33] 刘杉杉,黄秀芬,姜良,等.利用细菌人工染色体技术构建表达鹦鹉热衣原体PmpD基因的重组HVT[A].中国畜牧兽医学会家畜传染病学分会第八届全国会员代表大会暨第十五次学术研讨会论文集[C].2013. LIU S S,HUANG X F,JIANG L,et al.Construction of recombinant HVT expressing PmpD gene of Chlamydia psittaci using bacterial artificial chromosome technology[A].The 8th National Congress of the Livestock Infectious Diseases Branch of Chinese Association of Animal Science and Veterinary Medicine and the 15th Academic Seminar Papers Collection[C].2013.(in Chinese)
[34] IQBAL M.Progress toward the development of polyvalent vaccination strategies against multiple viral infections in chickens using Herpesvirus of turkeys as vector[J].Bioengineered,2012,3(4):222-226.
[35] 李延鹏,皮向成.一种共表达NDV HN和IBDV VP2基因的HVT的构建及其应用[P].CN107099513B,2020-07-14. LI Y P,PI X C.Construction and application of an HVT that expresses the NDV HN and IBDV VP2 genes[P].CN107099513B,2020-07-14.(in Chinese)
[36] 孙燕.表达传染性喉气管炎病毒gB、gD基因的重组火鸡疱疹病毒疫苗毒株构建及其免疫效力试验[D].扬州:扬州大学,2015. SUN Y.Construction and protective efficacy of two recombinant Herpesvirus of turkey vaccine strains expressing glycoprotein B and D of Infectious laryngotracheitis virus[D].Yangzhou:Yangzhou University,2015.(in Chinese)
[37] 钞安军.表达PCV2 CAP蛋白的重组猪伪狂犬病病毒的构建及小鼠免疫效力观察[D].郑州:河南农业大学,2013. CHAO A J.Construction of recombinant Pseudorabies virus expressing CAP protein of PCV2 and its immune efficacy in mice[D].Zhengzhou:Henan Agricultural University,2013.(in Chinese)
[38] 毛汐语,周雪珂,殷鑫欢,等.表达PEDV S和PoRV VP7蛋白的重组伪狂犬毒株构建[J].微生物学通报,2019,46(12):3345-3354. MAO X Y,ZHOU X K,YIN X H,et al.Construction of recombinant Pseudorabies virus strain expressing PEDV S and PoRV VP7 protein[J].Microbiology China,2019,46(12):3345-3354.(in Chinese)
[39] 夏德利.表达PCV2-Cap蛋白的TK^-/gE^-/gG^-重组伪狂犬病毒的构建及其免疫原性研究[D].北京:中国农业科学院,2017. XIA D L.Construction and immunogenicity of TK^-/gE^-/gG^- recombinant Pseudorabies virus expressing Porcine circovirus type 2 capsid protein[D].Beijing:Chinese Academy of Agricultural Sciences,2017.(in Chinese)
[40] 陈瑾.表达高致病性猪蓝耳病病毒(HuN4株)GP5重组伪狂犬病毒的构建及鉴定[D].北京:中国农业科学院,2008. CHEN J.Construction and identification of the recombination Pseudorabies virus expressing GP5 of highly pathogenic Porcine reproductive and respiratory syndrome virus (HuN4)[D].Beijing:Chinese Academy of Agricultural Sciences,2008.(in Chinese)
[41] TONG W,ZHENG H,LI G,et al.Recombinant Pseudorabies virus expressing E2 of Classical swine fever virus (CSFV) protects against both virulent Pseudorabies virus and CSFV[J].Antiviral Research,2020,173:104652.
[42] TONG W,LI G X,LIANG C,et al.A live,attenuated Pseudorabies virus strain JS-2012 deleted for gE/gI protects against both classical and emerging strains[J].Antiviral Research,2016,130:110-117.
[43] 王波,赵玉博,胡玉珍,等.表达2.3.2.1d分支H5亚型禽流感病毒HA基因重组鸭瘟病毒的构建和评价[J].中国预防兽医学报,2020,42(12):1262-1267. WANG B,ZHAO Y B,HU Y Z,et al.Construction and evaluation on recombinant Duck enteritis virus expressing HA gene of H5 subtype Avian influenza virus of clade 2.3.2.1d[J].Chinese Journal of Preventive Veterinary Medicine,2020,42(12):1262-1267.(in Chinese)
[44] 陈柳,余斌,倪征,等.表达小鹅瘟病毒VP2蛋白重组鸭瘟病毒的构建及其生物学特性[J].中国农业科学,2016,49(14):2813-2821. CHEN L,YU B,NI Z,et al.Construction and characterization of a recombinant Duck enteritis virus expressing VP2 gene of Goose parvovirus[J].Scientia Agricultura Sinica,2016,49(14):2813-2821.(in Chinese)
[45] DING L,CHEN P,BAO X,et al.Recombinant Duck enteritis viruses expressing the Newcastle disease virus (NDV) F gene protects chickens from lethal NDV challenge[J].Veterinary Microbiology,2019,232:146-150.
[46] LIU J X,CHEN P C,JIANG Y P,et al.Recombinant Duck enteritis virus works as a single-dose vaccine in broilers providing rapid protection against H5N1 influenza infection[J].Antiviral Research,2013,97(3):329-333.
[47] LIU J X,CHEN P C,JIANG Y P,et al.A Duck enteritis virus-vectored bivalent live vaccine provides fast and complete protection against H5N1 Avian influenza virus infection in ducks[J].Journal of Virology,2011,85(21):10989-10998.
[48] 胡玉珍,陈普成,丁蕾蕾,等.鸭瘟病毒疫苗株基因组复制非必需区的筛选[J].中国预防兽医学报,2018,40(10):897-901. HU Y Z,CHEN P C,DING L L,et al.Selection of nonessential regions for viral replication of Duck enteritis virus (vaccine strain) genome[J].Chinese Journal of Preventive Veterinary Medicine,2018,40(10):897-901.(in Chinese)
[49] 赵玉博,陈普成,胡玉珍,等.鸭瘟病毒疫苗株UL44.5和UL44基因间隔区作为复制非必需区的鉴定[J].中国预防兽医学报,2019,41(8):795-799. ZHAO Y B,CHEN P C,HU Y Z,et al.Identification of UL44.5 and UL44 gene spacers of Duck plague virus vaccine strains as non-essential regions for viral replication[J].Chinese Journal of Preventive Veterinary Medicine,2019,41(8):795-799.(in Chinese)
[50] SUN Y,YANG C H,LI J P,et al.Construction of a recombinant Duck enteritis virus vaccine expressing hemagglutinin of H9N2 Avian influenza virus and evaluation of its efficacy in ducks[J].Archives of Virology,2017,162(1):171-179.
[51] SHAO Y H,SUN J F,HAN Z X,et al.Recombinant Infectious laryngotracheitis virus expressing Newcastle disease virus F protein protects chickens against Infectious laryngotracheitis virus and Newcastle disease virus challenge[J].Vaccine,2018,36(52):7975-7986.
[52] 魏潇.表达新城疫病毒F蛋白传染性喉气管炎病毒gC缺失株的构建及评价[D].北京:中国农业科学院,2020. WEI X.Construction and evaluation of gC deletion Infectious laryngotracheitis virus strain expressing Newcastle disease virus F protein[D].Beijing:Chinese Academy of Agricultural Sciences,2020.(in Chinese)
[53] LI K,LIU Y,LIU C,et al.Effects of different promoters on the protective efficacy of recombinant Marek's disease virus type 1 expressing the VP2 gene of Infectious bursal disease virus[J].Vaccine,2016,34(47):5744-5750.
[54] ŚMIETANKA K,TYBOROWSKA J,OLSZEWSKA-TOMCZYK M,et al.A recombinant Turkey herpesvirus expressing F and HN genes of Avian avulavirus-1(AAvV-1) genotype Ⅵ confers cross-protection against challenge with virulent AAvV-1 genotypes Ⅳ and Ⅶ in chickens[J].Viruses,2019,11(9):784.
[55] 许曾焜.表达新城疫病毒F蛋白重组火鸡疱疹病毒的构建及免疫原性初步评价[D].北京:中国农业科学院,2020. XU Z K.Construction and preliminary immunogenicity evaluation of recombinant HVT expressing Newcastle disease virus F protein[D].Beijing:Chinese Academy of Agricultural Sciences,2020.(in Chinese)
[56] FENG Z H,CHEN J H,LIANG W W,et al.The recombinant Pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice[J].Virology Journal,2020,17(1):180.
[57] ZHENG H H,WANG L Q,FU P F,et al.Characterization of a recombinant Pseudorabies virus expressing Porcine parvovirus VP2 protein and porcine IL-6[J].Virology Journal,2020,17(1):19.
[58] NIU Y,LIU B,SUN C,et al.Construction of the recombinant Duck enteritis virus delivering capsid protein VP0 of the Duck hepatitis A virus[J].Veterinary Microbiology,2020,249:108837.
[59] 刘金华,刘立涛,孙洪磊.一种表达H9N2亚型禽流感病毒H9蛋白的重组火鸡疱疹病毒[P].CN109402071B,2021-04-27. LIU J H,LIU L T,SUN H L.A recombinant Turkey herpes virus expressing H9 protein of H9N2 subtype Avian influenza virus[P].CN109402071B,2021-04-27.(in Chinese)
[60] 亓文宝,苏冠铭,廖明,等.表达H7N9亚型高致病性禽流感病毒HA蛋白的重组火鸡疱疹病毒的构建与应用[P].CN110218706B,2021-03-16. QI W B,SU G M,LIAO M,et al.Construction and application of recombinant Turkey herpes virus expressing HA protein of H7N9 subtype Highly pathogenic avian influenza virus[P].CN110218706B,2021-03-16.(in Chinese)
[61] 蒋桃珍,张勇,宋亚芬,等.一种表达传染性法氏囊病病毒VP2蛋白重组病毒载体的构建与应用[P].CN111909965A,2020-11-10. JIANG T Z,ZHANG Y,SONG Y F,et al.Construction and application of a recombinant viral vector of the Infectious bursal disease virus VP2 protein[P].CN111909965A,2020-11-10.(in Chinese)
[62] 毕建敏.表达鸡新城疫病毒F基因的重组火鸡疱疹病毒的构建及其应用[P].CN106929483B,2020-09-25. BI J M.The construction and application of recombinant Turkey herpes virus to express the F gene of Newcastle disease virus[P].CN106929483B,2020-09-25.(in Chinese)
[63] 陈柳,张存,倪征,等.表达鸭坦布苏病毒E蛋白的重组鸭瘟病毒及其构建方法和应用[P].CN105039268A,2015-11-11. CHEN L,ZHANG C,NI Z,et al.Recombinant Duck plague virus expressing Duck Tembusu virus E protein and its construction method and application[P].CN105039268A,2015-11-11.(in Chinese)
[64] 杨承槐,李俊平,侯力丹,等.H5亚型禽流感重组鸭肠炎病毒rDEVΔUL2-H5株的构建及其应用[P].CN106995806A8,2019-08-30. YANG C H,LI J P,HOU L D,et al.Construction and application of recombinant Duck enteritis virus rDEVΔUL2-H5 strain of H5 subtype Avian influenza[P].CN106995806A8,2019-08-30.(in Chinese)
[65] 陈化兰,柳金雄,陈普成,等.表达分泌型鸭坦布苏病毒M/E蛋白的重组鸭病毒性肠炎病毒疫苗的构建和应用[P].CN103667197A,2014-03-26. CHEN H L,LIU J X,CHEN P C,et al.Construction and application of recombinant Duck viral enteritis virus vaccine expressing secreted Duck Tambusu virus M/E protein[P].CN103667197A,2014-03-26.(in Chinese)
[66] 陈化兰,柳金雄,步志高,等.表达禽流感病毒血凝素(HA)基因的重组鸭病毒性肠炎病毒疫苗株(rDEVus78Ha)及其构建方法和应用[P].CN102559610A,2012-07-11. CHEN H L,LIU J X,BU Z G,et al.Recombinant Duck viral enteritis virus vaccine strain (rDEVus78Ha) expressing Avian influenza virus hemagglutinin (HA) gene and its construction method and application[P].CN102559610A,2012-07-11.(in Chinese)
[67] 徐高原,张华伟,周明光,等.一种伪狂犬病病毒TK、gE、gI和gG基因缺失株及其制备方法和应用[P].CN112280753A,2021-01-29. XU G Y,ZHANG H W,ZHOU M G,et al.A Pseudorabies virus TK,gE,gI and gG gene deletion strain and its preparation method and application[P].CN112280753A,2021-01-29.(in Chinese)
[68] 陈红英,杨明凡,王林青,等.一种表达PPV VP2基因和猪IL-18基因的重组猪伪狂犬病毒株[P].CN105368796A,2016-03-02. CHEN H Y,YANG M F,WANG L Q,et al.A recombinant Porcine pseudorabies virus strain expressing PPV VP2 gene and porcine IL-18 gene[P].CN105368796A,2016-03-02.(in Chinese)
[69] 陈红英,崔保安,钞安军,等.表达猪圆环病毒Ⅱ型ORF2基因的重组猪伪狂犬病毒毒株及其制备方法[P].CN103509761A,2014-01-15. CHEN H Y,CUI B A,CHAO A J,et al.Recombinant Porcine pseudorabies virus strain expressing Porcine circovirus type Ⅱ ORF2 gene and its preparation method[P].CN103509761A,2014-01-15.(in Chinese)
[70] 马静云,符军,郑瑶瑶,等.表达猪繁殖与呼吸综合征病毒GP3/GP5/M基因的重组伪狂犬病毒、构建方法和应用[P].CN111875678A,2020-11-03. MA J Y,FU J,ZHENG Y Y,et al.Construction method and application of recombinant Pseudorabies virus expressing Porcine reproductive and respiratory syndrome virus GP3/GP5/M gene[P].CN111875678A,2020-11-03.(in Chinese)
[71] 王继春,张传健,郭仕琦,等.猪流行性腹泻病毒伪狂犬病病毒载体重组病毒、制备方法及其应用[P].CN112481223A,2021-03-12. WANG J C,ZHANG C J,GUO S Q,et al.Preparation method and application of Porcine epidemic diarrhea virus recombinant Pseudorabies virus[P].CN112481223A,2021-03-12.(in Chinese)
[72] 刘胜旺,邵昱昊,韩宗玺,等.一种表达新城疫病毒F蛋白的传染性喉气管炎重组病毒株及其构建方法和应用[P].CN111041004A,2020-04-21. LIU S W,SHAO Y H,HAN Z X,et al.Construction method and application of Infectious laryngotracheitis recombinant virus strain that expressing Newcastle disease virus F protein[P].CN111041004A,2020-04-21.(in Chinese)