新孢子虫GRA6表达、定位及对小鼠免疫效力的初步分析

doi:10.11843/j.issn.0366-6964.2015.04.016

Abstract

Abstract:

A variety of proteins are released during protozoan parasites invasion and growth in the host cell.Dense Granule protein is one of the key proteins which form and modify the parasitophorous vacuolar (PV).GRA6 gene was cloned from Neospora caninum genomic DNA.We constructed prokaryotic expression vector pGEX-GRA6 and eukaryotic expression vector pcDNA-GRA6.The protein was purified and immunized BALB/c mice to obtained high titer of serum.Results of indirect immunofluorescence (IFA) showed that GRA6 located at parasitophorous vacuolar network structure (PVN).BALB/c mice were immunized with recombinant protein or eukaryotic expression vector and challenged with Neospora caninum (2×10⁶).We monitored clinical symptoms and detected antibody levels of mice before each immunization.Thirty days after infection，brain burden of Neospora caninum was detected by RT-PCR.There were significant difference between immunization groups and control group (P<0.05)，which indicated that NcGRA6 had good immunoprotection for mice in both recombinant protein and eukaryotic expression vector.

CLC Number:

S852.723

XING Ming，LIU Jing，WANG Hui，HAO Pan，LIU Qun. Neospora caninum GRA6 Recombinant Expression，Localization and Preliminary Analysis on the Mice Immune Effect[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(4): 624-630.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2015.04.016

https://www.xmsyxb.com/EN/Y2015/V46/I4/624

References

［1］刘晶，余劲术，刘群，等.新孢子虫dNcSRS2重组蛋白间接ELISA的建立及其应用［J］.畜牧兽医学报，2006，37(10)：1036-1041.
LIU J，YU J S，LIU Q，et al.Establishment of recombinant dNcSRS2 protein based indirect ELISA for detection of antibody against Neospora caninum and its application［J］.Acta Veterinaria et Zootechnica Sinica，2006，37(10)：1036-1041.(in Chinese)
［2］DUBEY J P，SCHARES G，ORTEGA-MORA L M.Epidemiology and control of neosporosis and Neospora caninum ［J］. Clin Microbiol Rev，2007，20(2):323-367.
［3］刘群.新孢子虫病［M］.北京:中国农业大学出版社，2013.
LIU Q.Neosporosis［M］.Beijing：China Agricultural University Press，2013.(in Chinese)
［4］ZHANG W，DENG C，LIU Q，et al.First identification of Neospora caninum infection in aborted bovine foetuses in China ［J］.Vet Parasitol，2007，149 (1-2)：72-76.
［5］NAM H W.GRA proteins of Toxoplasma gondii：maintenance of host-parasite interactions across the parasitophorous vacuolar membrane ［J］.Korean J Parasitol，2009，47(Suppl)：S29-S37.
［6］ATKINSON R A，RYCE C，MILLER C M，et al.Isolation of Neospora caninum genes detected during a chronic murine infection ［J］.Int J Parasitol，2001，31(1)：67-71.
［7］ELLIS J T，RYCE C，ATKINSON R，et al.Isolation，characterization and expression of a GRA2 homologue from Neospora caninum ［J］.Parasitology，2000，120(Pt4)：383-390.
［8］LALLY N，JENKINS M，LIDDELL S，et al.A dense granule protein (NCDG1) gene from Neospora caninum ［J］.Mol Biochem Parasitol，1997，87(2)：239-243.
［9］LIDDELL S，LALLY N C，JENKINS M C，et al.Isolation of the cDNA encoding a dense granule associated antigen (NCDG2) of Neospora caninum ［J］.Mol Biochem Parasitol，1998，93(1)：153-158.
［10］LIU G，CUI X，HAO P，et al.GRA 14，a novel dense granule protein from Neospora caninum ［J］.Acta Biochim Biophys Sin（Shanghai），2013，45(7)：607-609.
［11］ASAI T，HOWE D K，NAKAJIMA K，et al.Neospora caninum：Tachyzoites express a potent type-I nucleoside triphosphate hydrolase［J］.Exp parasitol，1998，90(3)：277-285.
［12］COLLANTES-FERNÁNDEZ E，ZABALLOS A，ALVAREZ-GARCÍA G，et al.Quantitative detection of Neospora caninum in bovine aborted fetuses and experimentally infected mice by real-time PCR ［J］.J Clin Microbiol，2002，40(4)：1194-1198.
［13］NISHIKAWA Y，ZHANG H，IKEHARA Y，et al.Immunization with oligomannose-coated liposome-entrapped dense granule protein 7 protects dams and offspring from Neospora caninum infection in mice ［J］.Clin Vaccine Immunol，2009，16(6)：792-797.
［14］ELLIS J，MILLER C，QUINN H，et al.Evaluation of recombinant proteins of Neospora caninum as vaccine candidates (in a mouse model) ［J］.Vaccine，2008，26(47)：5989-5996.
［15］FAZAELI A，CARTER P E，DARDE M L，et al.Molecular typing of Toxoplasma gondii strains by GRA6 gene sequence analysis ［J］.Int J Parasitol，2000，30(5)：637-642.
［16］RAMAMOORTHY S，SANAKKAYALA N，VEMULAPALLI R，et al.Prevention of vertical transmission of Neospora caninum in C57BL/6 mice vaccinated with Brucella abortus strain RB51 expressing N.caninum protective antigens ［J］.Int J Parasitol，2007，37(13)：1531-1538.

[1]	XIAO Jie, LUO Yuejun, CHEN Hao, PU Jiayan, HE Wei, XIONG Changming, HAO Ge, REN Yongjun, YANG Guangyou. Screening of PCR-detection Targets of Eimeria stiedae, E. intestinalis, and E. magna in Rabbits and Establishment of the Single and Multiplex Direct PCR Methods [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4327-4337.
[2]	ZHENG Ruoyu, XIAO Jie, BAI Xin, CHEN Hao, PU Jiayan, REN Yongjun, YANG Guangyou. Evaluation of the Immunoprotective Effect of Recombinant Protein of Eimeria magna 3-Phosphoglyceraldehyde Dehydrogenase on Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4338-4349.
[3]	ZHENG Ruoyu, REN Yongjun, XIAO Jie, BAI Xin, PU Jiayan, CHEN Hao, YANG Guangyou. Evaluation of the Immunoprotective Effect of Recombinant Protein of Eimeria stiedae 3-Phosphoglyceraldehyde Dehydrogenase on Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2581-2595.
[4]	WANG Liyue, FENG Qianqian, CAI Weimin, LIU Dandan, HOU Zhaofeng, KANG Xilong, ZHANG Zhizhi, FAN Xuelian, ZHU Yu, XU Jinjun, PAN Zhiming, TAO Jianping. Cloning, Expression and Localization of Transcription Factor ApiAP2 of Eimeria necatrix [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4379-4388.
[5]	ZHAO Baocai, ZHAO Shuaiyang, GUAN Guiquan, LUO Jianxun, CAO Tianxing, ZHANG Zhigang, SHI Miao, LIU Junlong, ZHAO Hongxi. Transcriptome-based Analysis of the Effects of Theileria annulata on Host Cell Transformation-related Genes [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4398-4409.
[6]	YIN Yanling, HUANG Shuang, YAO Qian, WU Jiangping, GUO Haochen, YANG Xin, SONG Junke, ZHAO Guanghui. The Mechanisms of Circular RNA ciRS-7 Affecting the Propagation of Cryptosporidium parvum in HCT-8 Cells via Targeting miR-219a-5p [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3989-3999.
[7]	XIAO Ke, CHEN Ting, ZHAO Qiping, ZHU Shunhai, DONG Hui, LIU Manyu, YU Yu, HUANG Bing, HAN Hongyu. Preliminary Study on the Characteristics and Functions of HD Dome-containing Proteins in Eimeria tenella [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2608-2620.
[8]	YE Zhuang, WANG Lele, WANG Feiyan, LIU Yue, PENG Yuemei, SU Shijie, HOU Zhaofeng, XU Jinjun, TAO Jianping, LIU Dandan. Procaryotic Expression and Immunolocalization Analysis of the EnOXIO1 of Eimeria necatrix [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1553-1561.
[9]	JIAO Yuzhou, YANG Huanhuan, LU Yao, SHI Congcong, XU Xiaojuan, CAI Xuwang. Effect of Enterococcus faecium on Growth Performance and Intestinal Barrier of Broilers Infected with Eimeria tenella [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1210-1219.
[10]	LUO Yuejun, REN Yongjun, BAI Xin, CHEN Hao, PU Jiayan, ZHENG Ruoyu, XIAO Jie, YANG Guangyou. Preliminary Observation on the Immunoprotective Effects of Recombinant Surface Antigens SAG13 and SAG14 of Eimeria stiedae in Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 883-893.
[11]	LIANG Shanshan, ZHU Shunhai, ZHAO Qiping, HUANG Bing, DONG Hui, YU Yu, WANG Qingjie, YU Shuilan, WANG Haixia, HAN Hongyu. Preliminary Study on the Function of Elongation Factor 1α of Eimeria tenella [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(12): 3111-3121.
[12]	SONG Ruiqi, Huercha, ZHAI Xuejie, FAN Xinli, LI Min, ZHANG Mengyuan, SONG Jingjing, HAIRE Arman, KAMALI Wulijiang, GAILIKE Bayinchahan. Prokaryotic Expression of the Cysteine Proteinase Gene of Xinjiang Strain of Theileria equi/Babesia caballi and Its Bioinformatics Analysis [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(10): 2547-2556.
[13]	YANG Shoushen, SUN Xiaoshuang, QIU Yunfei, YANG Fei, TANG Jie, LIAN Wenjian, HUANG Cuiqin, CHIU Hungchuan. Differential Proteomic Analysis between Sporulated Oocyst and Unsporulated Oocyst of Cystoisospora suis [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(9): 2216-2226.
[14]	WANG Lu, ZHU Shunhai, ZHAO Qiping, HUANG Bing, HAN Hongyu, LIU Guiling, LI Zhihang, ZHAO Huanzhi, DONG Hui. Effects of Host Glyceraldehyde-3-phosphate Dehydrogenase on Eimeria tenella Sporozoites Invasion of Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 2088-2096.
[15]	WANG Panju, FAN Xinli, ZHANG Mengyuan, SONG Jingjing, LI Min, WU Lijiang, Bayin Chahan. Preparation and Application of Monoclonal Antibody against Truncated Bc48 of Babesia caballi Local Strains [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(9): 1882-1887.

Neospora caninum GRA6 Recombinant Expression，Localization and Preliminary Analysis on the Mice Immune Effect

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments