检测鸡TRIM25蛋白的免疫组化方法

doi:10.11843/j.issn.0366-6964.2018.04.027

Abstract

Abstract:

The experiment was conducted to set up the immunohistochemitry assay to detect chicken TRIM25 expression in the chicken's tissues. Chicken TRIM25 gene was amplified from chicken's spleen and the gene was cloned into a prokaryotic expression vector pEASY-Blunt E1 to construct recombinant plasmid named as pEASY-Blunt E1-TRIM25. The recombinant plasmid was expressed in E. coli and then identified by SDS-PAGE and Western blot. After being purified, the fusion protein was inoculated to rabbits and BALB/c mice as antigen for preparing polyclonal antibody. The titer of the antibody was detected by ELISA, and the sera antibodies were used to setup immunohistochemistry assay; the tissue sections of chicken's lung and spleen and liver were assayed. The results showed that TRIM25 was expressed as fusion protein with 48 kD. The titers of the antibody in rabbits' and mice's sera were above 1:10⁶. The results of immunohistochemistry assay indicated that TRIM25 mainly exists in the cytoplasm of lymphocytes, hepatocyte, and epithelial cells in chicken's lung, spleen and liver. In conclusion, the immunohistochemitry assay mediated with polyclonal antibody against chicken TRIM25 were successfully set up in this study and TRIM25 distribution in some tissues of chickens was firstly reported.

CLC Number:

S852

CHU Ying, MA Wen-jie, HU Wei-guo, CHEN Ru, QIU Jian-hua, GUO Hui-jun, LI Hong-mei. Immunohistochemistry Assay for Detecting Chicken TRIM25 in Different Tissues[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(4): 865-870.

0
/ / Recommend

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

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2018.04.027

https://www.xmsyxb.com/EN/Y2018/V49/I4/865

References

[1] OZATO K, SHIN D M, CHANG T H, et al. TRIM family proteins and their emerging roles in innate immunity[J]. Nat Rev Immunol, 2008, 8(11):849-860.
[2] DESHAIES R J, JOAZEIRO C A P. RING domain E3 ubiquitin ligases[J]. Ann Rev Biochem, 2009, 78:399-434.
[3] SONG B, GOLD B, O'HUIGIN C, et al. The B30.2(SPRY) domain of the retroviral restriction factor TRIM5α exhibits lineage-specific length and sequence variation in primates[J]. J Virol, 2005, 79(10):6111-6121.
[4] URANO T, SAITO T, TSUKUI T, et al. Efp targets 14-3-3σ for proteolysis and promotes breast tumour growth[J]. Nature, 2002, 417(6891):871-875.
[5] UCHIL P D, QUINLAN B D, CHAN W T, et al. Trim E3 ligases interfere with early and late stages of the retroviral life cycle[J]. PLoS Pathog, 2008, 4(2):e16.
[6] ZHANG S J, GUO J T, WU J Z, et al. Identification and characterization of multiple TRIM proteins that inhibit hepatitis B virus transcription[J]. PLoS One, 2013, 8(8):e70001.
[7] GACK M U, ALBRECHT R A, URANO T, et al. Influenza A virus NS1 targets the ubiquitin ligase TRIM25 to evade recognition by the host viral RNA sensor RIG-I[J]. Cell Host Microbe, 2009, 5(5):439-449.
[8] RAJSBAUM R, ALBRECHT R A, WANG M K, et al. Species-specific inhibition of RIG-I ubiquitination and IFN induction by the influenza A virus NS1 protein[J]. PLoS Pathog, 2012, 8(11):e1003059.
[9] INN K S, GACK M U, TOKUNAGA F, et al. Linear ubiquitin assembly complex negatively regulates RIG-I-and TRIM25-mediated type i interferon induction[J]. Mol Cell, 2011, 41(3):354-365.
[10] FENG Z Q, CHENG Y, YANG H L, et al. Molecular characterization, tissue distribution and expression analysis of TRIM25 in Gallus gallus domesticus[J]. Gene, 2015, 561(1):138-147.
[11] ZHANG D D, LI H M, ZHANG Z S, et al. Antibody responses induced by recombinant ALV-A gp85 protein vaccine combining with CpG-ODN adjuvant in breeder hens and the protection for their offspring against early infection[J]. Antiviral Res, 2015, 116:20-26.
[12] 刘庆祥. TRIM家族蛋白对I型干扰素信号通路的调控作用[J]. 中山大学研究生学刊:自然科学、医学版, 2015, 36(4):42-52.
LIU Q X. Functions of TRIM family proteins in type I interferon signaling pathway[J]. Journal of the Graduates Sun Yat-Sen University:Natural Sciences, Medicine, 2015, 36(4):42-52. (in Chinese)
[13] 刘海港, 李宏梅, 王树迎, 等. 山羊β雌激素受体多克隆抗体的制备及免疫组化方法的建立[J]. 畜牧兽医学报, 2013, 44(2):302-308.
LIU H G, LI H M, WANG S Y, et al. Preparation of polyclonal antibody against goat estrogen receptor β and its application in immunohistochemistry assay[J]. Acta Veterinaria et Zootechnica Sinica, 2013, 44(2):302-308. (in Chinese)

[1]	CHONG Qian, ZHAO Xuehui, CAO Qing, XUE Huiwen, GOU Huitian. Research Progress of Applied Research on Listeria monocytogenes Phage [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 1819-1826.
[2]	HAN Fuzhen, CAI Limeng, LI Zhuoran, WANG Xueying, XIE Weichun, KUANG Hongdi, LI Jiaxuan, CUI Wen, JIANG Yanping, LI Yijing, SHAN Zhifu, TANG Lijie. Research Progress on the Mechanism of Intestinal Flora-Mediated Regulation of Intestinal Mucosal Immunity by Secondary Bile Acids and Their Receptors [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 1904-1913.
[3]	HUANG Jin, LI Siyuan, MAO Li, CAI Xuhang, XIE Lingling, WANG Fu, ZHOU Hua, LI Jizong, LI Bin. Eukaryotic Expression of Bovine Coronavirus S1 Protein and Establishment and Application of Indirect ELISA [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2050-2060.
[4]	HAN Yang, GUAN Shuaiyin, LI Zhen, ZHOU Saisai, YUAN Honggen, SONG Yunfeng. Prokaryotic Expression and Protein Activity of Porcine Circovirus Type 3 Rep [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2061-2071.
[5]	SONG Xiaoqing, DENG Ruide, LI Xin, LI Jiao, LI Runcheng, DU Lifei, DONG Wei, GE Meng. Establishment of ELISA for Detection of PCV4-Cap Antibody and Sero-epidemiological Survey [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2072-2079.
[6]	ZHOU Yang, WU Weizi, CAO Weisheng, WANG Fuguang, XU Xiuqiong, ZHONG Wenxia, WU Liyang, YE Jian, LU Shousheng. A Whole Genome Sequencing Method for African Swine Fever Virus based on Nanopore Sequencing Technology was Established [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2080-2089.
[7]	MA Rumeng, ZHAO Yuliang, MA Mingshuang, GUO Guihai, LIU Xinzi, LI Jiaxuan, CUI Wen, JIANG Yanping, SHAN Zhifu, ZHOU Han, WANG Li, QIAO Xinyuan, TANG Lijie, WANG Xiaona, LI Yijing. Comparative Study on the Immune Response Induced by the Different Porcine Receptor Bacteria with Expressing the Protective Antigen S1 of Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2090-2099.
[8]	XU Hong, SHANG Hongqi, ZHANG Xue, QIAN Jiali, WANG Chuanhong, SONG Xu, BAO Meiying, LIU Shiyu, ZHANG Gege, GUO Rongli, ZHAO Yongxiang, FAN Baochao, LI Bin. Inhibition Effect of C8orf4 Gene Encoding Protein on in vitro Replication of Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2100-2108.
[9]	XIONG Ting, HE Xianming, ZHAO Xiya, ZHUANG Tingting, HUANG Meizhen, LIANG Shijin, YU Chuanzhao, LIANG Xuejing, CHEN Ruiai. Whole Genome Analysis of Three Predominant Epidemic Strains of Chicken Infectious Bronchitis Virus and Their Pathogenicity [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2109-2122.
[10]	XU Zhenyu, DENG Xiaoyu, WANG Yueli, SUN Can, WU Aodi, CAO Jian, YI Jihai, WANG Yong, WANG Zhen, CHEN Chuangfu. Biological Characteristics of Brucella abortus A19ΔBtpA Deletion Strain and Its Immunogenicity Study [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2135-2145.
[11]	ZHAO Canqi, FENG Yu, LÜ Lang, LI Yanjun, WEI Yulei, DING Jiabo, CHEN Xiang, JIANG Hui. Study on Purification of Bovine Brucellosis by Competitive ELISA and Indirect ELISA [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2146-2153.
[12]	LUO Ting, HAN Zhu, XU Yefen, CAI Lin, SUOLANG Sizhu, XU Jinhua, NIU Jiaqiang. Whole Genome Sequencing and Sequence Analysis on T10 of Mycoplasma bovis Strain from Yaks in Xizang [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2154-2167.
[13]	GAO Jie, LI Xiaocheng, MU Yang, ZHANG Hui, WEI Rong, LI Jie. Biological Characteristics and Immune Effect Evaluation of Outer Membrane Vesicles of Capsular Type B Pasteurella multocida [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2168-2175.
[14]	WANG Jing, ZHANG Shujuan, HU Xia, LIU Xiangyang, ZHANG Xingcui, SONG Zhenhui. CD44 Regulates Na⁺/H⁺ Exchanger 3 Activity by Influencing Porcine Epidemic Diarrhea Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2176-2185.
[15]	ZHENG Rui, LIU Zishi, ZHANG Kangyou, YAN Yong, WEI Ling, ZEREN Wengmu, DINGZE Demi, HUANG Jianjun, WANG Li, WEI Yong. Isolation, Identification and Biological Characterization of Colletotrichum jasminigenum in Stems of Peanuts [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2206-2213.

Immunohistochemistry Assay for Detecting Chicken TRIM25 in Different Tissues

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments