tva受体基因起始密码子突变对鸡感染A亚群禽白血病病毒的影响  

Effect of initiation codon mutation within tva receptor gene on chicken resistance to infection by avian leukemia virus subgroup A

作　　者：陈伟国 邝智祥 张翔宇 李文雪 徐慧娟 朱学凤 谢青梅[1] CHEN Weiguo;KUANG Zhixiang;ZHANG Xiangyu;LI Wenxue;XU Huijuan;ZHU Xuefeng;XIE Qingmei(College of Animal Science,South China Agricultural University/Guangdong Provincial Key Laboratory of Animal Health Aquaculture and Environmental Control,Guangzhou 510642,China;Guangdong Love-Health Biotechnology Co.,Ltd.,Qingyuan 511800,China)

机构地区：[1]华南农业大学动物科学学院/广东省畜禽健康养殖与环境控制重点实验室,广东广州510642 [2]广东爱健康生物科技有限公司,广东清远511800

出　　处：《华南农业大学学报》2023年第3期367-373,共7页Journal of South China Agricultural University

基　　金：广东省重点领域研发计划(2020B020222001);国家重点研发计划(2021YFD1300100);清远市科技计划项目(2021DZX019)。

摘　　要：[目的]探索tva受体基因起始密码子突变(tva c.3G>A)对鸡感染A亚群禽白血病病毒(Avian leukemia virus subgroup A, ALV-A)的影响。[方法]利用Sanger测序和RT-PCR验证我国黄羽肉鸡存在tva c.3G>A突变。利用流式细胞术检测tva c.3G>A突变对鸡体外感染RCASBP(A)-GFP荧光报告病毒的影响。通过ALV-A体内攻毒试验,探究tva c.3G>A突变对鸡体内感染ALV-A的影响。利用直接测序方法对我国黄羽肉鸡品系tva c.3G>A突变位点进行基因分型。[结果]Sanger测序和RT-PCR结果鉴定我国黄羽肉鸡品系tva基因编码区第3位碱基由G突变为A,该突变引起tva基因起始密码子序列由ATG突变为ATA。流式细胞术检测结果显示野生型tva c.3G/G鸡胚成纤维细胞(Chicken embryo fibroblast, CEF)对RCASBP(A)-GFP易感,纯合突变型tva c.3A/A CEF抗RCASBP(A)-GFP感染,表明tva c.3G>A突变导致鸡体外抗RCASBP(A)-GFP的感染。ALVA体内攻毒试验的结果表明,tva c.3G>A突变导致鸡体内抗ALV-A的感染。tva c.3G>A突变位点基因分型发现,CB01、CB08、CB10和CB15品系存在纯合抗性基因型tva c.3A/A,频率分别为0.10、0.15、0.23和0.08。[结论]tva c.3G>A突变引起鸡在体外、体内抗ALV-A感染,tva c.3G>A突变位点可作为ALV-A的遗传抗性位点。【Objective】To explore the effect of initiation codon mutation within tva receptor gene(tva c.3G>A)on resistance of chickens to infection by avian leukemia virus subgroup A(ALV-A).【Method】Sanger sequencing and RT-PCR were used to verify the presence of tva c.3G>A mutation in Chinese yellow-feathered broilers.The effect of tva c.3G>A mutation on infection of chickens by RCASBP(A)-GFP fluorescence reporter virus in vitro was detected using flow cytometry.The effect of tva c.3G>A mutation on infection of chickens by ALV-A was investigated using ALV-A challenge test in vivo.Direct sequencing was used to genotype tva c.3G>A mutation site within Chinese yellow-feathered broiler lines.【Result】The results of Sanger sequencing and RT-PCR identified the mutation from G to A on the third base in the coding region of tva gene of Chinese yellow-feathered broilers,which caused the mutation from ATG to ATA in the initial codon sequence of tva gene.The result of flow cytometry showed that chicken embryo fibroblasts(CEFs)of wild-type tva c.3G/G were susceptible to infection by RCASBP(A)-GFP,while the homozygous mutant tva c.3A/A CEFs were resistant to infection by RCASBP(A)-GFP,indicating that tva c.3G>A mutation led to chicken resistance to infection by RCASBP(A)-EGFP in vitro.The results of ALV-A challenge test in vivo also indicated that tva c.3G>A mutation led to chicken resistance to infection by ALV-A.Genotyping of tva c.3G>A revealed that homozygous resistance genotype tva c.3A/A was present in lines CB01,CB08,CB10 and CB15,with the frequencies of 0.10,0.15,0.23 and 0.08,respectively.【Conclusion】The tva c.3G>A mutation causes chicken resistance to infection by ALV-A in vitro and in vivo,and the tva c.3G>A mutation site can be used as the genetic resistance site of ALV-A.

关 键 词：A亚群禽白血病病毒 tva受体基因 起始密码子 突变 遗传抗性 黄羽肉鸡 

分 类 号：S855.3[农业科学—临床兽医学]