全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

鸡α干扰素基因的克隆、原核表达及抗病毒活性测定

DOI: 10.7668/hbnxb.2009.01.010, PP. 40-43

Keywords: ,&alpha,干扰素,克隆,原核表达,抗病毒活性

Full-Text   Cite this paper   Add to My Lib

Abstract:

通过PCR技术从罗曼鸡肝脏基因组中扩增鸡α干扰素(ChIFN-α)全基因,并克隆和测序。序列分析表明,ChIFN-α基因全长为582bp,亚克隆其成熟蛋白编码基因489bp,利用基因重组技术构建了重组质粒,使ChIFN-α置于原核表达载体pQE30的T5启动子下并同6×His(多聚组氨酸标签)-Tag融合。经酶切和PCR鉴定,DNA测序证实重组质粒pQEChIFN-α构建正确将pQEChIFN-α转化大肠杆菌M15感受态细胞,用IPTG诱导表达。SDS-PAGE和Westernblot证实表达出分子质量为19.80kDa的融合蛋白。表达的蛋白以包涵体形式存在,经变性、复性处理后,在Vero细胞上抗水泡性口炎病毒的活性为1.16×103U/mg。本研究成功表达了ChIFN-α,表达的重组蛋白具有一定的生物活性。

References

[1]  1 陆承平.兽医微生物「Ml.北京:中国农业出版社,2001: 452.
[2]  2 Sekellick M J, Ferrandino A F, Flopkins D A,et al. Chicken inteeron gene:cloning,expression,and analysis [J].Journal of lnteiteron and(Zokine Research,1994,14 (2):71一79.
[3]  3 汪明,吴志光,夏春.肉鸡IFN alp ha基因的克隆、 序列分析以及在大肠杆菌中的表达[J].农业生物技术 学报,2000,8 (4):337一381.
[4]  4 夏春,汪明,朱凌云,等.惠阳胡须鸡r-a基因克 隆和序列分析[J].畜牧兽医学报,2000,31 (6):563 - 565.
[5]  5 夏春,汪明,杨琪,等.丝羽乌骨鸡正、基因克隆 和序列分析[J].中国免疫学杂志,2000,16 (9):508 - 513.
[6]  6 Takehara K,Itobayashi K,Ruttanapumma R,et al. Adjuvant ei3ect of chicken inte}eron-gamma for inactivated Salmonella Fnteritidis antigen[J].Journal of Veterinary Medical Science, 2003,65(12):1337一1341.
[7]  7 C,Liu J,Wu Z G, et al. The intetferon-alpha genes from three chicken lines and its etJects on H9N2 influenza viruses [J].Animal Biotechnology,2004,15 (1):77一88.
[8]  8 Rauw F, Lambrecht B,van den Berg T. Pivotal role of ChIFNgamma in the patla genesis and immunosuppression of infectious bursal disease [J].Avian Pathology,2007,36 (5): 367一374.
[9]  9 Rubinstein S, Familletti P C, Pestka S. Convenient assay for intetferons [J].Journal of Virology,1981,37(2):755一758.
[10]  10 Haghighi H R, Abdul-Careem M F, Dara R A,et cal. tokine gene expression in chicken cecal tonsils following treatment with probiotics and Salmonella infection[J].Vet- erinary 1VTicrobiology,2008,126(1一3):225一233.
[11]  11 Pfeffer L M,Dinarello C A,Herberman R B,et al. Biological properties of recombinant alpha-inte ferons:40th anniversary of the discovery of inte ferons [J].Cancer Research,1998, 58(12):2489一2499.
[12]  12 Pestka S,Trotta P P. Recombinant alpha-inteferons from naturally occurring genes[J].Introduction Seminars in On- cology,1997,24 (3 Suppl 9):1一3.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133