| 新城疫F_(48)E_9株L基因克隆与鉴定 |
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| 引用本文： | 闫丽辉,曹殿军,刘培欣,孙建宏.新城疫F_(48)E_9株L基因克隆与鉴定[J].中国预防兽医学报,2001(4). |
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| 作者姓名： | 闫丽辉 曹殿军 刘培欣 孙建宏 |
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| 基金项目： | 国家重点基础研究发展规划项目 (G19990 1190 2 ) |
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| 摘 要： | 本实验根据已知新城疫病毒L基因序列分别设计了两对引物 ,引物 1(L1)、引物 2 (L2 )、引物 3(L3)、引物 4(L4)。以L1/L2为引物可以扩出 40 5 0bpF4 8E9株L基因的A片段 (LA) ,以L3/L4为引物可扩增出 35 0 4bpF4 8E9株L基因的B片段 (LB)。LA和LB 2个片段经特异性双酶切后用T4DNA连接酶连接成完整的F4 8E9株L基因并克隆到pMD18_T载体中 ,对克隆后L基因 5’端、3’端和连接点处的核苷酸序列进行了测定 ,经DNASIS软件分析表明为NDVL基因 ,证明我们获得了F4 8E9株L基因
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| 关 键 词： | 新城疫病毒 L基因 克隆 鉴定 |
Cloning and Identification of NDV F_(48) E_9 Strain L Gene |
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| YAN Lihui,CAO Dianjun ,LIU Peixin,SUN Jianhong.Cloning and Identification of NDV F_(48) E_9 Strain L Gene[J].Chinese Journal of Preventive Veterinary Medicine,2001(4). |
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| Authors: | YAN Lihui CAO Dianjun LIU Peixin SUN Jianhong |
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| Institution: | YAN Lihui,CAO Dianjun *,LIU Peixin,SUN Jianhong |
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| Abstract: | cDNA encoding the L gene of F 48 E 9 strain was cloned in two steps.Two pairs of primers were synthesized according to the alien published NDV L gene sequence.The virion RNA was obtained from purifed NDV F 48 E 9 strian and used as a template for cDNA synthesis.Then cDNAs of A and B fragment of L gene were amplified by PCR and cloned into the pMD18_T Vector respectively.The positive clones of LA and LB were identified by molecular weight?restriction endonucleases digestion and PCR.According to restriction endonucleases digestion sites,the LA was cut and ligased with LB.The positive clones of L gene were identified by sequence analysis. |
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| Keywords: | NDV L gene Cloning Identification |
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