Cloning and Prokaryotic Expression of the F Gene of NDV La Sota Strain

doi:10.3321/j.issn:1000-7091.2006.03.027

Abstract

Abstract: A pair of primers was designed according to the sequence of F gene of NDV La Sotastrain(AF077761) for amplifying the F gene by RT-PCR.1 700 bp DNA fragment was amplified and cloned into pGEM-T easy vector.After restriction endonuclease analysis and sequencing,the fragment was inserted into the expression vector PET_32a.The recombinant plasmid produced a 83 kd fusion protein in E.coli BL21(DE3) under the induction of IPTG.The expression quantity and immunoreactivity of the fusion protein was detected by SDS_PAGE electrophoresis and Western blotting.

Key words: Newcastal disease virse (NDV), F gene, Clone, Prokaryotic expression

CLC Number:

S831.5

FU Fang, JIANG Bei-yu, ZHANG Li, GAO Xuan, ZHANG Biao. Cloning and Prokaryotic Expression of the F Gene of NDV La Sota Strain[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2006, 21(3): 117-120. doi: 10.3321/j.issn:1000-7091.2006.03.027.

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References

[1] 殷震,刘景华.动物病毒学(第二版)[M].北京:科学出版社,1997.
[2] J·萨姆布鲁克(美).金冬雁,黎孟枫,译.分子克隆(第二版)[M].北京:科学出版社,1996.55-56.
[3] De Leeuw O S,Hartog L,Koch G,et al.Effect of fusion protein cleavage sitemutations on virulence of New castle disease virus:nonvirulent cleavage site mutants revert to virulence after one passage in chicken brain[J].J Gen Viro 1,2003,84 (2):475-484.
[4] Peeters B P,De Leeuw O S,Koch G,et al.Rescue of New castle disease virus from cloned cDNA:evidence that cleavability of the fusion protein is a major determinant for virulence[J].J Virol,1999,73 (6):5001-5009.
[5] 吴艳涛,刘秀梵,张如宽,等.表达新城疫病毒F48E8株融合蛋白基因的重组鸡痘病毒及其免疫效力[J].生物工程学报,2000,16(5):592-594.
[6] 姜永厚,陈奖励.鸡新城疫病毒F基因和鸡IL-2重组DNA疫苗的构建[J].中国预防兽医学报,2001,23(2):81-83.
[7] 方维焕,梁雪芽.减毒沙门氏菌为载体在细胞中表达新城疫病毒融合蛋白[J].生物化学与生物物理学报,2002,43(4):488-493.
[8] Sakaguch I M,Nakamura H,Sonoda K,et al.Protection of chickens from New castle disease by vaccination with a linear plasmid DNA expressing the F protein of New castle disease virus[J].Vaccine,1996,14 (8):747-752.
[9] bruce S S,Daniel J K,Joyce D B.Characterition of New castle disease virus isolates by RT-PCR coupled to direct nucleotide sequencing and development of sequence database fo r pathotype predictionand molecular epidemiological analysis[J].J Clin Microbiol,1995,33(10):2624-2630.
[10] 许发芝,仲大莲,余为一.新城疫病毒(NDV)F基因的克隆及其真核表达重组质粒的构建[J].安徽农业大学学报,2003,30(3):280-284.
[11] 邓明俊,张彦明,梁荣,等.鸡新城疫病毒F基因高效真核表达质粒的构建[J].西北农林科技大学学报(自然科学版),2003,31(2):9-12.

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