中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 鸡痘病毒基因探针检测方法的建立及初步应用 中国预防兽医学报 2007,29 (2) 126-129
Position: Home > Articles > Application of gene probe for detection of fowl poxvirus infection Chinese Journal of Preventive Veterinary Medicine 2007,29 (2) 126-129

鸡痘病毒基因探针检测方法的建立及初步应用

作  者：
郭建顺;金宁一;张国才;张学东;冯立文;沈晓峰
单  位：
吉林大学畜牧兽医学院;军事医学科学院军事兽医研究所
关键词：
鸡痘病毒;4b核心蛋白基因;基因探针
摘  要：
利用PCR方法成功克隆了鸡痘病毒(Fowl poxvirus,FPV)4b核心蛋白基因549bp片段,序列分析表明,该序列与模板DNA(AF198100)碱基序列的同源性为99.45%,只有3个碱基差异,第215位由C→T,第386位由T→A,第388位由G→A。回收FPV 4b核心蛋白基因549bp片段,以其为模板制备了地高辛标记的DNA探针。对新标记的探针进行标记效率检测,结果显示其标记效率为100mg/L;敏感性检测表明,该探针对同源DNA的检出限量为10pg;特异性检测结果表明,用本试验所标记的探针对提取的FPV 282E4和FPV JL株DNA、重组质粒pMD18-T-4b进行检测结果均呈阳性,而鸡马立克氏病病毒、鸡传染性喉气管炎病毒、CEF的核酸提取物均成阴性,说明该探针具有较强的特异性。初步应用表明,本试验所建立的FPV的基因探针检测法可用于FPV的检测。
译  名：
Application of gene probe for detection of fowl poxvirus infection
作  者：
GUO Jian-shun1,2, JIN Ning-yi2*, ZHANG Guo-cai1, ZHANG Xue-dong1, FENG Li-wen1, SHEN Xiao-feng1 (1. College of Veterinary Medicine and Animal Science, Jilin University, Changchun 130062, China; 2. The Military Veterinary Institute, Academy of Military Medicine of PLA, Changchun 130062, China)
关键词：
FPV; 4b core protein gene; gene probe
摘  要：
A 549 bp DNA fragment derived from fowl poxvirus (FPV) 4b core protein gene was obtained by PCR and inserted into pMD18-T vector. Positive clones were identified by PCR and restriction digestion. The recombinant plasmid (pMD18-T-4b) was sequenced. Blast analysis showed the 4b core protein gene is highly conserved and the nucleotide homogeneity between the cloned sequence and the published DNA sequence of 4b gene of PFV (AF198100) was 99.45 %. Only 3 nucleotide differences were found between them, where C,T and G at position 215, 386 and 388 were substituted by T, A and A respectively. DNA robe was produced by labelling the PCR fragment with digoxigenin-dUTP. This probe was used to perform Dot-blot hybridization against DNA from FPV282E4, FPV JL, pMD18-T-4b, MDV, AILTV and DNA from CEF. Positive results were obtained for FPV282E4, FPV JL and pMD18-T-4b, but not for MDV, AILTV and from CEF. The probe was highly sensitive and could detect a minimal 10 pg homology DNA. The results indicated that the probe could be used to detect DNA extracted from the skin at the vFV282 inoculation site on day 1-7 post immunization.

相似文章

导出引用
计量
文章访问数: 22
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46