Recombinant Expression and Antibody Cross-reaction of the Outer Membrane Protein VP1008 and Ferric Vibrioferrin Receptor of <i>Vibrio parahaemolyticus</i>

LIANG Xiaxia; YUAN Qianyun; LIU Lei; GUO Shanshan; WANG Wenbin

doi:10.13386/j.issn1002-0306.2021030061

Volume 42 Issue 19

Sep. 2021

Turn off MathJax

Article Contents

Abstract

References

Science and Technology of Food Industry > 2021 > 42(19): 144-151. > DOI: 10.13386/j.issn1002-0306.2021030061

LIANG Xiaxia, YUAN Qianyun, LIU Lei, et al. Recombinant Expression and Antibody Cross-reaction of the Outer Membrane Protein VP1008 and Ferric Vibrioferrin Receptor of Vibrio parahaemolyticus[J]. Science and Technology of Food Industry, 2021, 42(19): 144−151. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021030061.

Citation:

PDF (15227 KB)

Recombinant Expression and Antibody Cross-reaction of the Outer Membrane Protein VP1008 and Ferric Vibrioferrin Receptor of Vibrio parahaemolyticus

LIANG Xiaxia^{1, 2, 3,},
YUAN Qianyun^{1, 2, 3},
LIU Lei^{1, 2, 3},
GUO Shanshan^{1, 2, 3},
WANG Wenbin^{1, 2, 3, ,}

1.
School of Food Science and Engineering, Jiangsu Ocean University, Lianyungang 222005, China
2.
Jiangsu Key Laboratory of Marine Bioresources and Environment, Lianyungang 222005, China
3.
Jiangsu Marine Biological Industry Technology Collaborative Innovation Center, Lianyungang 222005, China

More Information

Received Date: March 07, 2021
Available Online: August 05, 2021

Graphical Abstract

Abstract

Abstract

Objective: The study of V. parahaemolyticus specific surface antigens is of importance to develop immunoassays of immune detection. The immunogenicity study of V. parahaemolyticus differential outer membrane protein (VP1008) and ferric vibrioferrin receptor (pvuA) has not been revealed. Methods: The target genes were amplified from the genome DNA and the recombinant plasmids pET-28a(+) were respectively constructed and transformed into E. coli BL21. The two recombinant proteins were both induced by Isopropyl-β-D-Thiogalactoside (IPTG). After purified by a NI-NTA column, the purified recombinant proteins were immunized to the BALB/c mice. The cross-reaction of the polyclone antibodies (pAb) with different isolates of V. parahaemolyticus and other Vibrio species was studied by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Results: The recombinant protein VP1008 and Vibrio ferritin receptor were expressed as inclusion bodies, the pAb both strongly reacted to the corresponding recombinant protein (>1000 K), reacted with 7 strains of V. parahaemolyticus (4.5~13.5 K), and basically did not react with 10 strains of Vibrio, the cross-reaction with Enterobacteriaceae strains was related with the residual proteins of E. coli during the purification steps. The pAb also both reacted with the target protein in the cell lysates of the V. parahaemolyticus strains. Comparatively, the pAb against VP1008 had a higher antibody titer and immunogenicity. Conclusion: The pAb against the recombinant Omp VP1008 recognized the V. parahaemolyticus strains with higher titer, which laid the foundation for preparing a new immunoassay against V. parahaemolyticus.
- Vibrio parahaemolyticus,
- outer membrane protein,
- recombinant expression,
- ferric vibrioferrin receptor,
- immunogenicity,
- immunoassay

FullText(HTML)

References (34)

References

[1]	Craig B A, Jenkins C, Claire J, et al. Genomic epidemiology of domestic and travel-associated Vibrio parahaemolyticus infections in the UK, 2008–2018[J]. Food Control,2020,115:107244. doi: 10.1016/j.foodcont.2020.107244
[2]	张巍巍, 李颖, 王群, 等. 全基因组数据应用于多克隆副溶血弧菌感染爆发的病原学分析[J]. 疾病监测,2020,35(6):518−522. [Zhang W W, Li Y, Wang Q, et al. Application of whole genome sequencing in etiology investigation of an outbreak of polyclonal Vibrio parahaemolyticus infection[J]. Disease Surveillance,2020,35(6):518−522. doi: 10.3784/j.issn.1003-9961.2020.06.013
[3]	李梦飞, 明红霞, 樊景凤, 等. 近岸及河口环境中副溶血弧菌的生态适应性及其主要的毒力基因分布特征[J]. 生命科学,2019,31(2):209−215. [Li M F, Ming H X, Fan J F, et al. Ecological adaptability and distribution features of main virulence gene of Vibrio Parahaemolyticus in the nearshore and estuarine environments[J]. Chinese Bulletin of Life Sciences,2019,31(2):209−215.
[4]	刘国胜, 董峰光, 潘丽芳, 等. 我国淡水产品中致病性弧菌的分布及其特征的研究进展[J]. 食品安全质量检测学报,2019,10(3):9−14. [Liu G S, Dong F G, Pan L F, et al. Review on the distribution and characteristics of pathogenic Vibrio in freshwater products in China J]. Journal of Food Safety and Quality,2019,10(3):9−14.
[5]	Taiwo M, Baker-Austin C, Craig P, et al. Comparison of toxR and tlh based PCR assays for Vibrio parahaemolyticus[J]. Food Control,2017,77(1):116−120.
[6]	刘斌. 海水养殖鱼类主要细菌病多联疫苗的研制及应用[D]. 青岛: 中国海洋大学, 2007. Liu B. Development and application on polyvalent vaccine of bacterial pathogens of marine fish[D]. Qing Dao: Ocean University of China, 2007.
[7]	Jadeja R, Janes M E, Simonson J G. Development of rapid and sensitive anti-flagellar monoclonal antibody based lateral flow device for the detection of Vibrio vulnificus from oyster homogenate[J]. Food Control,2015,56:110−113. doi: 10.1016/j.foodcont.2015.03.019
[8]	高璐, 欧阳敏, 张辉, 等. 胁迫生长条件下副溶血性弧菌的生物特征分析[J]. 食品科学,2018,39(6):177−182. [Gao L, Ouyang M, Zhang H, et al. Biological characteristics of Vibrio parahaemolyticus during growth in adverse environment[J]. Food Science,2018,39(6):177−182. doi: 10.7506/spkx1002-6630-201806028
[9]	毛芝娟. 副溶血性弧菌Vibrio parahaemolyticus主要外膜蛋白的克隆、表达和免疫原性研究[D]. 杭州: 浙江大学, 2007. Mao Z J. Cloning, expression and immunogenicity of major outer membrane proteins of Vibrio parahaemolyticus[D]. Hangzhou: Zhejiang University, 2007.
[10]	Chen Z X, Chu X, Wang S N, et al. Six genes of ompA family shuffling for development of polyvalent vaccines against Vibrio alginolyticus and Edwardsiella tarda[J]. Fish and Shellfish Immunology,2018,75(7):308−315.
[11]	张丽芳, 罗薇. 外膜蛋白的结构、功能及其应用的研究进展[J]. 中国畜牧兽医, 2014, 41(10): 55-61. Zhang L F, Luo W. Preparation and immune protection of the fusion protein FlaA-OmpK vaccine of Vibrio parahaemolyticus[J]. Journal of Fishery Sciences of China, 2014, 19(05): 848−853.
[12]	毛芝娟, 由振强, 魏永伟, 等. 副溶血性弧菌ZJ2003株两种铁调外膜蛋白的克隆、表达和免疫原性[J]. 中国水产科学,2007,14(4):563−569. [Mao Z J, You Z Q, Wei Y W, et al. Cloning, expression and immunogenicity of two outer membrane proteins of Vibrio parahaemolyticus ZJ2003 strain[J]. Fisheries Science in China,2007,14(4):563−569. doi: 10.3321/j.issn:1005-8737.2007.04.006
[13]	Hui L, Ming Zhi Y, Peng B, et al. Immunoproteomic identification of polyvalent vaccine candidates from Vibrio parahaemolyticus outer membrane proteins[J]. Journal of Proteome Research,2010,9(5):2573−2583. doi: 10.1021/pr1000219
[14]	丁承超, 谢曼曼, 曾海娟, 等. 弧菌外膜蛋白Ompk基因序列分析及其多克隆抗体的制备[J]. 食品科学,2016,37(23):141−146. [Ding C C, Xie M M, Zeng H J, et al. Gene sequence analysis and polyclonal antibody preparation of outer membrane protein, Ompk, in Vibrio[J]. Food Science,2016,37(23):141−146. doi: 10.7506/spkx1002-6630-201623024
[15]	李小飞, 李槿年, 季晶晶, 等. 拟态弧菌外膜蛋白OmpU基因的原核表达及其免疫保护性研究[J]. 中国水产科学,2008,15(2):307−312. [Li X F, Li J N, Ji J J, et al. Study on prokaryotic expression of outer membrane protein OmpU gene of Vibrio mimicus and its immune protection[J]. Journal of Fishery Sciences of China,2008,15(2):307−312. doi: 10.3321/j.issn:1005-8737.2008.02.015
[16]	马振宁, 郑磊, 林海美, 等. 抗OmpW单克隆抗体的制备与特性鉴定[J]. 细胞与分子免疫学杂志,2011,56(9):982−984, 988. [Ma Z N, Zheng L, Lin H M, et al. Preparation and characterization of monoclonal antibody against OmpW[J]. Journal of Fishery Sciences of China,2011,56(9):982−984, 988.
[17]	Cheng Z X, Chu X, Wang S N, et al. Six genes of OmpA family shuffling for development of polyvalent vaccines against Vibrio alginolyticus and Edwardsiella tarda[J]. Fish and Shellfish Immunology,2018,75:308−315. doi: 10.1016/j.fsi.2018.02.022
[18]	Funahashi T, Moriya K, Uemura S, et al. Identification and characterization of pvuA, a gene encoding the ferric vibrioferrin receptor protein in Vibrio parahaemolyticus[J]. Journal of Bacteriology,2002,184(4):936−946. doi: 10.1128/jb.184.4.936-946.2002
[19]	Tomotaka T, Tatsuya F, Noriyuki O, et al. The Vibrio parahaemolyticus pvuA1 gene (formerly termed psuA) encodes a second ferric vibrioferrin receptor that requires tonB2[J]. Fems Microbiology Letters,2011,324(1):73−79. doi: 10.1111/j.1574-6968.2011.02389.x
[20]	Yamamoto S, Akiyama T, Okujo N, et al. Demonstration of a ferric vibrioferrin-binding protein in the outer membrane of Vibrio parahaemolyticus[J]. Microbiology and Immunology,1995,39(10):759−766. doi: 10.1111/j.1348-0421.1995.tb03268.x
[21]	Wang W B, Liu J X, Guo S S, et al. Identification of Vibrio parahaemolyticus and Vibrio Spp. specific outer membrane proteins by reverse vaccinology and surface proteome[J]. Frontiers in Microbiology,2021,11:625315. doi: 10.3389/fmicb.2020.625315
[22]	Yang J, Anishchenko I, Park H, et al. Improved protein structure prediction using predicted interresidue orientations PNAS[J]. 2020, 117 (3): 1496−1503.
[23]	张丹阳, 吕育财, 田红毅, 等. 枯草芽孢杆菌β 甘露聚糖酶的克隆表达及重组酶性质研究[J]. 食品工业科技,2020,41(6):88−105. [Zhang D Y, Lv Y C, Tian H Y, et al. Cloning, expression and characterization of recombinant β-mannanase from Bacillus subtilis[J]. Science and Technology of Food Industry,2020,41(6):88−105.
[24]	黄伦辉, 鲍会静, 张嘉懿, 等. 禽蛋黄过敏源Gal d 6的重组表达、纯化鉴定[J]. 食品工业科技,2020,41(13):118−127. [Huang L H, Bao H J, Zhang J Y, et al. Recombinant expression, purification and identification of egg yolk allergen Gal d 6[J]. Science and Technology of Food Industry,2020,41(13):118−127.
[25]	何雨峰, 韦胜, 徐泽, 等. 红曲菌Mn-SOD基因克隆、表达及序列分析[J]. 食品工业科技,2020,41(18):81−86. [He Y F, Wei S, Xu Z, et al. Cloning, expression and sequence analysis of Mn-SOD gene of Monascus[J]. Science and Technology of Food Industry,2020,41(18):81−86.
[26]	程晋霞, 曾静, 张蕾, 等. 霍乱弧菌鞭毛蛋白单克隆抗体的制备及双抗体夹心ELISA的建立[J]. 细胞与分子免疫学杂志,2013,29(11):1169−1173. [Cheng J X, Zeng J, Zhang L, et al. Preparation of monoclonal antibodies against flagellin core protein of Vibrio cholerae and its application in establishing double-antibody sandwich ELISA for testing Vibrio cholerae from food products[J]. Journal of Cell and Molecular Immunology,2013,29(11):1169−1173.
[27]	高云山, 桑雨浓, 梁夏夏, 等. 弧菌外膜蛋白OmpK多肽免疫原制备及其免疫原性研究[J]. 江苏海洋大学学报,2013,29(11):1169−1173. [Gao Y S, Sang Y N, Liang X X, et al. Preparation and immunogenicity study of Vibrio conserved peptide immunogens from OmpK[J]. Journal of Jiangsu Ocean University,2013,29(11):1169−1173.
[28]	李碧华. 革兰氏阴性菌外膜蛋白的结构、功能及遗传学[J]. 微生物与感染, 1991 (5): 193-196. Li B H. Structure, function and genetics of outer membrane proteins in gram negative bacteria[J]. Microorganism and Infection, 1991(5): 193−196.
[29]	戴启宇, 宋蓓, 王会英, 等. AIF 多克隆抗体的制备纯化和鉴定[J]. 实用医药杂志,2009,26(10):67−68. [Dai Q Y, Song B, Wang H Y, et al. Preparation, purification and characterization of polyclonal antibody of apoptosis-inducing factor (AIF)[J]. Journal of Practical Medicine,2009,26(10):67−68. doi: 10.3969/j.issn.1671-4008.2009.10.042
[30]	Phianphak W, Rengpipat S, Rukpratanporn S, et al. Production of monoclonal antibodies for detection of Vibrio harveyi[J]. Diseases of Aquatic Organisms,2005,63(2-3):161−168.
[31]	伦镜盛, 张设熙, 董亚萍, 等. 弧菌外膜蛋白OmpU的免疫交叉反应性和交叉保护性[J]. 微生物学报,2016,56(5):867−879. [Lun J S, Zhang S X, Dong Y P, et al. Immune cross-reactivity and cross-protection of outer membrane protein OmpU in Vibrio[J]. Acta Microbiologica Sinica,2016,56(5):867−879.
[32]	Klemm P, Vejborg R M, Hancock V. Prevention of bacterial adhesion[J]. Appl Microbiol Biotechnol,2010,88(2):451−459. doi: 10.1007/s00253-010-2805-y
[33]	Kietzman C C, Gao G, Mann B, et al. Dynamic capsule restructuring by the main pneumococcal autolysin LytA in response to the epithelium[J]. Nature Communications,2016,7(1):10859. doi: 10.1038/ncomms10859
[34]	王亚磊, 王权, 白雪瑞, 等. 副溶血性弧菌SH112 株OmpA 蛋白的高效表达及免疫学特性[J]. 微生物学报,2019,59(10):1937−1947. [Wang Y L, Wang Q, Bai X R, et al. High expression and immunological characteristics of OmpA protein in Vibrio parahaemolyticus SH112 strain[J]. Acta Microbiologica Sinica,2019,59(10):1937−1947.