猪白细胞介素18基因的克隆及真核表达重组质粒的构建

doi:10.16431/j.cnki.1671-7236.2017.09.007

Abstract

Abstract:

To obtain recombinant eukaryotic expression plasmid of porcine interleukin-18(IL-18), the whole gene of porcine IL-18 gene was amplified from porcine spleen, lung and lymph nodes by RT-PCR and cloned into eukaryotic expression vector pZJ-1. The recombinant expression plasmid pZJ-IL-18 were identified by enzyme digestion and sequencing analysis, and was transfected into 293T cells.The expression of IL-18 was detected by RT-PCR and Western blotting in both gene and protein levels. The results showed that the eukaryotic expression plasmid of porcine IL-18 was constructed and could express transiently in 293T cells. Western blotting result confirmed that porcine IL-18 polyclonal antibody could react specifically with approximately 17 ku expression products,and indicated that IL-18 could express correctly and be responsive.This study constructed the eukaryotic expression plasmid of porcine IL-18 gene which could express transiently in 293T cells, and laid the foundation for studying function of IL-18.

Key words: porcine; IL-18 gene; eukaryotic expression vector; clone

CLC Number:

SHI Ang, SHI Qing-he, LI Xin-guo, WANG Yu-guo, YANG Li, LI Shuang-shuang, CHEN Lu, WANG Chuan-qing. Cloning and Eukaryotic Expression Plasmid Construction of Porcine Interleukin-18 Gene[J]. China Animal Husbandry and Veterinary Medicine, 2017, 44(9): 2587-2592.

References

[1] Okamura H, Kashiwamura S, Tsutsui H. Regulation of interferon-gamma production by IL-12 and IL-18[J]. Current Opinion in Immunology, 1998, 10(3):259.
[2] Yan Y, Niu L, Deng J, et al. Adjuvant effects of recombinant giant panda (Ailuropoda melanoleuca) IL-18 on the canine distemper disease vaccine in mice[J]. Journal of Veterinary Medical Science, 2015, 77(2):187.
[3] 郭晓庆, 郑兰兰, 付朋飞, 等. 表达PCV2 CAP蛋白和猪IL-18重组PRV PGO18株的生物学特性[J]. 中国兽医学报, 2016, 36(5):707-711.
[4] 王春岩, 刘莉, 杨文涛,等. 猪白细胞介素18成熟蛋白基因的克隆与表达[J]. 中国微生态学杂志, 2012, 24(9):769-772.
[5] 沈国顺, 金宁一, 秦晓光, 等. 表达PRRS病毒GP5、GP3和猪IL-18的核酸疫苗的构建及实验免疫研究[J]. 免疫学杂志, 2006, 22(6):629-634.
[6] Okamura H, Tsutsi H, Komatsu T. Cloning of a new cytokine that induces IFN-gamma production by T cells[J]. Nature, 1995, 378(6552):88.
[7] 郑敏, 金宁一, 张洪勇,等. 猪白细胞介素18成熟蛋白基因的克隆及在大肠杆菌中的表达[J]. 中国兽医学报, 2003, 23(5):430-432.
[8] 韩丽, 何潇湘, 王瑞宁,等. 猪IL-18增强猪细小病毒灭活疫苗猪体细胞免疫效果的研究[J]. 河南农业大学学报, 2016, 2:224-228.
[9] 周红莉, 郭丽, 王健伟,等. 粘膜免疫佐剂研究进展[J]. 中国生物工程杂志, 2006, 26(3):83-88.
[10] 余协中, 王红宁, 黄勇,等. 细胞因子类佐剂在黏膜免疫中的作用[J]. 动物医学进展, 2007, 28(8):30-33.
[11] Muneta Y, Shimoji Y, Yokomizo Y. Molecular cloning of porcine interleukin-1beta converting enzyme and differential gene expression of IL-1beta converting enzyme, IL-1beta, and IL-18 in porcine alveolar macrophages[J]. Journal of Interferon & Cytokine Research, 1999, 19(11):1289-1296.
[12] 马全英, 祝秀梅, 安芳兰,等. 白细胞介素18研究进展[J]. 贵州畜牧兽医, 2014, 38(2):21-24.
[13] 孙文长. 白细胞介素18研究进展[J]. 国际检验医学杂志, 2000, 21(2):57-59.
[14] 何淑雅, 薛志红, Ming Z,等. 人白细胞介素18(hIL-18)的cDNA基因克隆及序列测定[J]. 中国现代医学杂志, 2001, 11(4):30-32.
[15] 张瑞军, 田志刚, 魏海明,等. 重组人白细胞介素18的纯化及其生物学活性研究[J]. 中国免疫学杂志, 2001, 17(8):411-414.
[16] 张盼盼, 何静, 康银峰,等. 猪α干扰素与白介素-18融合基因的克隆、原核表达及其抗病毒活性的初步研究[J]. 中国畜牧兽医,2014,41(6):29-33.
[17] Pirhonen J. Regulation of IL-18 expression in virus infection[J]. Scandinavian Journal of Immunology, 2001, 53(6):533-539.
[18] Zheng L L, Guo X Q, Zhu Q L, et al. Construction and immunogenicity of a recombinant pseudorabies virus co-expressing porcine circovirus type 2 capsid protein and interleukin 18[J]. Virus Research, 2015, 201:8-15.
[19] Matsui K, Yoshimoto T, Tsutsui H. Propionibacterium acnes treatment diminishes CD4⁺ NK1.1+ T cells but induces type Ⅰ T cells in the liver by induction of IL-12 and IL-18 production from Kupffer cells[J]. Journal of Immunology, 1997, 159(1):97-106.
[20] Micallef M J, Ohtsuki T, Kohno K. Interferon-gamma-inducing factor enhances T helper 1 cytokine production by stimulated human T cells:Synergism with interleukin-12 for interferon-gamma production[J]. European Journal of Immunology, 1996, 26(7):1647-1651.
[21] Trzeciak M, Sokolowska-Wojdyol M, Barans'ka-Rybak W. Interleukin 18-A pleiotropic cytokine involved in the Th1 and Th2 immunological response[J]. Postepy Dermatologii I Alergologii, 2011, 28(4):309-312.
[22] Kinoshita M, Kuranaga N, Matsumoto A. Multiple interleukin-18 injections promote both mouse Th1 and Th2 responses after sublethal Escherichia coli infection[J]. Clinical & Experimental Immunology, 2006, 143(1):41-49.
[23] Rahman M M, Uyangaa E, Han Y W. Modulation of systemic and mucosal immunity against an inactivated vaccine of Newcastle disease virus by oral co-administration of live attenuated Salmonella enterica serovar Typhimurium expressing chicken interleukin-18 and interferon-α[J]. Journal of Veterinary Medical Science, 2015, 77(4):395.
[24] Tovey M G, Lallemand C. Adjuvant activity of cytokines[J]. Methods in Molecular Biology, 2010, 626(626):287-309.
[25] Lihsiang H, Li H P, Yiyang L, et al. Adjuvant effects of chicken interleukin-18 in avian Newcastle disease vaccine[J]. Vaccine, 2010, 28(5):1148-1155.
[26] 景志忠, 窦永喜, 侯俊琳,等. 猪白介素-18基因的克隆及其序列分析[J]. 中国兽医科学, 2004, 34(7):3-7.
[27] 段永兰, 赵丽. 河南长白猪白细胞介素18基因的克隆及其在大肠杆菌中的表达[J]. 黑龙江畜牧兽医, 2012, 9:52-54.

Cloning and Eukaryotic Expression Plasmid Construction of Porcine Interleukin-18 Gene

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