坏死梭杆菌外膜蛋白基因的克隆与表达

›› 2013, Vol. 40 ›› Issue (6): 57-60.

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Cloning and Expression of Outer Membrane Proteins (OMP) of Fusobacterium necrophorum (AB)

XU Jing^1,2, CHEN Li-zhi², LIU Xiao-ying², FENG Er-kai², WANG Sun-jie^1,2, CAO Yue²

1. School of Biology and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212018, China;
2. State Key Laboratory for Molecular Biology of Special Economical Animals, Institute of Special Animal and Plant Science, Chinese Academy of Agricultural Sciences, Changchun 130112, China

Received:2012-11-30 Online:2013-06-20 Published:2013-06-20

Abstract

Abstract: The outer membrane protein (OMP) was deposited in GenBank. The upstream and downstream primers were designed, and OMP gene was amplified by PCR. Then the target gene was inserted into the vector pMD18-T Simple and pET-28a, and the recombinant vectors were used to select and be transformed into E.coli TOP10 and BL21 (DE3), respectively. The plasmid from E.coli TOP10 and BL21 (DE3) was digested by restricted endonuclease enzyme of XhoⅠand EcoRⅠ. Then the plasmid of enzyme digestion was analyzed by its sequence. The recombinant protein was expressed by IPTG induction. We successfully obtained OMP by identification of SDS-PAGE, and result of Western blotting showed that the OMP had immune activity.

Key words: Fusobacterium necrophorum; outer membrane protein (OMP); induction expression; immune activity

CLC Number:

Q786

XU Jing, CHEN Li-zhi, LIU Xiao-ying, FENG Er-kai, WANG Sun-jie, CAO Yue. Cloning and Expression of Outer Membrane Proteins (OMP) of Fusobacterium necrophorum (AB)[J]. , 2013, 40(6): 57-60.

References

1 元英芳,黄瑛,杜艳芬,等.牛分枝杆菌外膜蛋白基因ompA的原核表达及免疫活性分析[J]. 中国动物传染病学报,2009,17(4): 43~46.
2 余波,徐景峨,王璇,等. 鸡肠炎沙门氏菌ompA基因的克隆与表达[J].中国畜牧兽医,2012,39(12): 20~23.
3 陈立志. 牛源致病性坏死梭杆菌分离鉴定及其白细胞毒素免疫特性研究.长春:吉林大学,2008.
4 Brazier J S. Human infections with Fusobacterium necrophorum[J]. Anaerobe, 2006,12(4):165~172.
5 Cao Y, Johnson H M,Bazemore-Walker C R,et al. Improved enrichment and proteomic identification of outer membrane proteins from a gram-negative bacterium: Focus on Caulobacter crescentus[J]. Proteomics, 2012,12(2): 251~262.
6 Checkley S L, Janzen E D, Campbell J R, et al. Efficacy of vaccination against Fusobacterium necrophorum infection for control of liver abscesses and footrot in feedlot cattle in Western Canada[J]. Can Vet J, 2005, 46(11):1002~1007.
7 Kaplan C W, Ma X Y, Paranjpe A, et al. Fusobacterium nucleatum outer membrane proteins Fap2 and RadD induce cell death in human lymphocytes[J]. Infect Immun, 2010, 78(11):4773~4778.
8 Khushiramani R, Girisha S K, Karunasagar I, et al. Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish[J]. Protein Expr Purif, 2007, 51(2):303~307.
9 Korn A, Rajabi Z, Wassum B,et al. Enhancement of uptake of lipopolysacharide in macrophaes by the major outer membrane protein OmpA of gram-negative bacteria[J]. Infection Immunology, 1995, 63(7): 2697~2705.
10 Liu P F, Shi W Y, Zhu W H, et al. Vaccination targeting surface FomA of Fusobacterium nucleatum against bacterial co-aggregation: Implication for treatment of periodontal infection and halitosis[J]. Vaccine, 2010, 28(19):3496~3505.
11 Roy S, Das A B,Ghosh A N,et al. Purification, pore-forming ability, and antigenic relatedness of the major outer membrane protein of Shigella dysenteriae type-1[J]. Infection Immunology, 1994, 62(10): 4333~4338.
12 Takada H, Ogawa T, Yoshimura F, et al. Immunobiological activities of a porin fraction isolated from Fusobacterium nucleatum ATCC 10953[J]. Infect Immun, 1988, 56(4):855~863.
13 Weiner J H, Li L. Proteome of the Escherichia coli envelope and technological challenges in membrane proteome analysis[J]. Biochimica et Biophysica Acta (BBA) Biomembranes, 2008, 1778(9): 1698~1713.
14 Zhou H T, Meyer K, Ganter M, et al. Identification of a Fusobacterium necrophorum isolate that contains a new variant of the leukotoxin gene (lktA) from the hoof of a sheep with ovine footrot[J]. Vet Microbiol, 2011, 149(3~4):524~525.

Cloning and Expression of Outer Membrane Proteins (OMP) of Fusobacterium necrophorum (AB)

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