Application of Several PCR Methods in Cloning Unknown Genes of Duck Enteritis Virus

Home > Archive>Volume 36, Issue 12, 2009 >1842-1848

Application of Several PCR Methods in Cloning Unknown Genes of Duck Enteritis Virus
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                        PAN Hua-QiPAN Hua-Qi
1. Institute of Applied Ecology, Chinese Academy Sciences, Shenyang, Liaoning 110016, China; Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture at Nanjing Agricultural University, Nanjing, Jiangsu 210095, China; College of
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CAO Rui-BingCAO Rui-Bing
Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture at Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
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CHEN Pu-YanCHEN Pu-Yan
Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture at Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
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LIU LeiLIU Lei
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu 730070, China
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WANG Shu-JinWANG Shu-Jin
Institute of Applied Ecology, Chinese Academy Sciences, Shenyang, Liaoning 110016, China
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PAN Guang-YanPAN Guang-Yan
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu 730070, China
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HU Jiang-ChunHU Jiang-Chun
Institute of Applied Ecology, Chinese Academy Sciences, Shenyang, Liaoning 110016, China
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Abstract:

Degenerate PCR, modified targeted gene walking PCR, modified thermal asymmetric interlaced PCR and Long-PCR were performed with genomic DNA of duck enteritis virus as a template and three fragments (5350 bp, 11083 bp, 2905 bp) of DEV genome were obtained. DNA sequence analysis revealed nine open reading frames encoding UL3, UL4, UL5, UL25, UL26, UL27, UL28, and UL30 gene of DEV, respectively. These sequences have been submitted to GenBank with accession numbers from EF554396 to EF554402. Joint application of several PCR techniques is efficient in cloning unknown genes of duck enteritis virus.

Key words:Duck enteritis virus, Unknown gene, Degenerate PCR, Thermal asymmetric interlaced PCR, Targeted gene walking PCR, Long-PCR

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PAN Hua-Qi, CAO Rui-Bing, CHEN Pu-Yan, LIU Lei, WANG Shu-Jin, PAN Guang-Yan, HU Jiang-Chun. Application of Several PCR Methods in Cloning Unknown Genes of Duck Enteritis Virus[J]. Microbiology China, 2009, 36(12): 1842-1848

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