猪圆环病毒2型Cap蛋白的截短表达与活性检测

Abstract

Abstract: Designing and synthesizying one pair of primers according to the published PCV2 genome sequence, a fragment of about 480 bp long was amplified by RT-PCR technique with specific primers. Then the target fragment was directionally cloned into pGEX-6p-1 vector. After identifying with enzyme cut and sequencing, the recombinant plasmid was transformed into E. coli BL21(DE3). The recombinant Cap protein was expressed in inclusion body form after induction with IPTG. After denaturation, renaturation of recombinant protein by affinity chromatography purification, purification of the recombinant protein concentration was 0.396 mg/mL. Western blotting and indirect ELISA testing showed that the purified recombinant protein retained better antigenicity and specificity.

Key words: porcine circovirus type 2; Cap protein; prokaryotic expression; activity detection

CLC Number:

S858.28

CAO Gui-zhong. Characterization of Porcine Circovirus Type 2 Truncated Cap Protein Expressed in E.coli[J]. .

References

1 马涛, 王先炜, 李玉峰, 等.不同佐剂对猪圆环病毒2型灭活抗原免疫效果的影响[J]. 中国兽医科学,2011,41(6):593~600.

2 王文秀, 沈志强, 吕素芳, 等. 猪圆环病毒2型ORF2与EGFP融合蛋白表达载体的构建及在鸭心肌细胞中的表达[J].中国兽医科学, 2011,41(10):1047~1051.

3 史小红, 徐志文, 郭万柱, 等. 融合表达猪圆环病毒2型ORF1和ORF2基因真核质粒的构建及对小鼠的免疫原性[J].中国兽医学报, 2011,31(9):1246~1251.

4 乔翠, 高凤山, 许崇波. 猪圆环病毒2型分子致病机理及相关疾病研究进展[J].中国畜牧兽医,2012,39(2):170~173.

5 曹彦琼, 王先炜, 李文良, 等. 猪圆环病毒2型Cap蛋白单克隆抗体的制备与鉴定[J]. 中国兽医科学, 2011, 41(9):911~916.

6 Jaganathan S, Toung O P, Yee P L, et al. Genetic characterization of porcine circovirus 2 found in Malaysia[J]. Virol J,2011, 8:437.

7 Liu J, Chen I, Du Q, et al. The ORF3 protein of porcine circovirus type 2 is involved in viral pathogenesis in vivo[J]. J Virol, 2006, 80(10):5065~5073.

8 Liu Q, Tikoo S K, Babiuk L A. Nuclear localization of the ORF2 protein encoded by porcine circovirus type 2[J]. Virology,2001, 285(1):91~99.

9 Lou Z, Li X, Li Z, et al. Expression and antigenicity characterization for truncated capsid protein of porcine circovirus type 2[J]. Can J Vet Res,2011,75(1):61~64.

10Magar R, Müller P, Larochelle R. Retrospective serological survey of antibodies to porcine circovirus type 1 and type 2[J]. Can J Vet Res, 2000, 64(3):184~186.

11Mahe D, Blanchard P, Truong C, et al. Differential recognition of ORF2 protein from type 1 and type 2 porcine circoviruses and identification of immunorelevant epitopes[J]. J Gen Virol,2000,81(7):1815~1824.

12McClenahan S D, Krause P R, Uhlenhaut C. Molecular and infectivity studies of porcine circovirus in vaccines[J]. Vaccine,2011, 29(29~30):4745~4753.

13Rodríguez-Arrioja G M, Segalés J, Balasch M, et al. Serum antibodies to porcine circovirus type 1 and type 2 in pigs with and without PMWS[J]. Vet Rec,2000, 146(26):762~764.

14Rosell C, Segalés J, Rovira A, et al. Porcine circovirosis in Spain[J]. Vet Rec,2000,146(20):591~592.

15Stadejek T, Podgorska K, Porowski M, et al. Linked outbreaks and control of porcine reproductive and respiratory syndrome and postweaning multisystemic wasting syndrome in a pig farm in Poland[J]. Vet Rec,2011, 169(17):441.

[1]	JIAO Linlin, CHENG Yuting, WU Qingguo, WU Shuang, WU Zhi, ZHU Shanyuan, QIAN Yingjuan. Prokaryotic Expression and Polyclonal Antibody Preparation of Duck Tembusu Virus Capsid Protein [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2395-2402.
[2]	LI Can, LI Chuchu, ZENG Wei, ZHANG Ning, JI Chunxiao, CHEN Tao. Recombinant Expression and Function Study of RegⅢγ Protein in Porcine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2414-2426.
[3]	SONG Yue, SU Shengjie, ZHANG Fan, BAI Fan, DAI Lingli, WANG Na, WANG Dawei, YANG Qianwen, ZHAO Shihua, ZHANG Yuemei. Prokaryotic Expression and Polyclonal Antibody Preparation of Mannheimia haemolytica Truncated Leukotoxin [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2479-2486.
[4]	ZHANG Fangyuan, LIN Yating, YANG Dawei, CHEN Hu, LI Guimei, SHAN Hu. Prokaryotic Expression of African Swine Fever Virus P30 Protein and Establishment of Indirect ELISA Detection Method [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2012-2022.
[5]	JIANG Haijun, WANG Dan, GUO Yu, WEI Li, HOU Lei, WANG Jing, ZHU Shanshan, LIU Jue. Fusion Expression and Immunogenicity of Recombinant Cap-flagellin Protein of Porcine Circovirus Type 3 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2044-2052.
[6]	HU Leyu, HU Xinyao, LI Ying, ZHAO Shengmiao, SHEN Fengzhen, WANG Changfa, LI Liangliang, WANG Tongtong, REN Huiying. Bioinformatics Analysis of HO-1 Gene and Preparation of Polyclonal Antibody in Dezhou Donkey [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1499-1510.
[7]	WANG Wenjie, RU Penghui, YU Chuan, DU Fuxi, CHEN Songbiao, SHANG Ke, ZHANG Chunjie, CHENG Xiangchao. Prokaryotic Expression of VP2 Protein of Luoyang Feline Panleukopenia Virus Isolate and Polyclonal Antibody Preparation [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1511-1521.
[8]	LIU Jiaxing, HAN Xueying, ZHANG Xinru, DENG Jiaxin, YAO Lunguang, LIU Yangkun. Preparation and Immunogenicity Analysis of Ferritin Nanoparticles Carrying Antigenic Epitopes of Porcine Epidemic Diarrhea Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1567-1574.
[9]	JIA Xinyue, MA Jing, ZHANG Yanyan, MA Xun, WANG Zhengrong, BO Xinwen. Bioinformatics Analysis and Prokaryotic Expression of EGR-07243 and EGR-07243 Genes of Echinococcus granulosus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 847-858.
[10]	LIU Xinyue, LI Lifang, WANG Zhenbao, ZHANG Wenliang, ZHANG Xin, ZHAO Hongtao, LI Pengyu, ZHENG Haotian, XIAO Jin, QI Peng. Adjuvants Screening and Immune Protection of Porcine Circovirus Type 2 Synthetic Peptide Vaccine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1068-1080.
[11]	ZHANG Zhitao, LI Xianglong, KONG Lingli, LUO Yuxin, WANG Dongying. Prokaryotic Expression of Serine/Threonine Protein Phosphatase 2A in Fasciola gigantica and Expression Analysis at Different Stages [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1107-1117.
[12]	GAN Lu, ZHENG Huizhen, NUO Mingdalai, LIU Yan, JIN Min, HE Wenwen, WEN Licui, LI Yongchang, GAILIKE·Bayinchahan. Propagation of Trypanosoma evansi Yili Strain and Cloning, Expression and Bioinformatics Analysis of Its PFR Gene [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 469-478.
[13]	ZHOU Wenfeng, ZHANG Yingjie, BAI Yihan, ZHANG Luhua, MAI Jinhui, WANG Naidong, YANG Yi, ZHAN Yang, WANG Dongliang. Complete Genome Sequence Analysis and Cap CTL Epitopes Prediction of Porcine Circovirus Type 2 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 656-665.
[14]	LYU Shuangfei, MA Xun, WANG Jing, KONG Cuilian, KOU Lijun, LIU Caixia, SHI Weidi, KANG Lichao, REN Huijie. Prokaryotic Expression and Immunoreactivity of PA-LF1 Fusion Protein of Bacillus anthracis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 674-683.
[15]	ZHANG Kai, ZHANG Chunping, GE Shengqiang, SUN Chunxi, CHENG Jie, FU Chunyu, WANG Gang, NIU Xing, PENG Jun. Expression of p22 Protein of African Swine Fever Virus,Preparation of Polyclonal Antibody and Establishment of Indirect ELISA Method [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 270-279.

Characterization of Porcine Circovirus Type 2 Truncated Cap Protein Expressed in E.coli

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments