中国2个地方品系猪SLA-3原核表达载体构建及表达

doi:10.16431/j.cnki.1671-7236.2015.02.009

Abstract

Abstract: To construct the prokaryotic expressing vector of SLA-3 derived from Hebao and Laiwu pigs (named as SLA-3-HB and SLA-3-LW) and to express the interest of protein,the extracellular domain of SLA-3 were amplified by PCR,and then they were cloned into the pMD19-T Simple vector followed by transforming into Escherichia coli Top10.By cleavage and sequencing,the positive clones were selected.After cleavage and purification of the recombinant plasmids,the interest of genes were further ligated to pET-21a(+) and transformed into the Escherichia coli BL21.The interest of genes were induced to express by IPTG and the interest of proteins were detected by SDS-PAGE.The results showed that the extracellular domain of SLA-3-HB and SLA-3-LW were successfully amplified with the molecular weight of 850 bp.It was also shown that the SLA-3-HB and SLA-3-LW were successfully cloned into pMD19-T Simple vector and the positive recombinant plasmids with correct sequences were selected.Further study proved that SLA-3-HB and SLA-3-LW were successfully ligated into pET-21a(+) with the 831 bp inserted fragments.After induction,SLA-3-HB and SLA-3-LW were all expressed successfully and the molecular weight of the interest of proteins were about 31 ku,and the contents of the expression of the proteins reached 40 percent.In this study,the prokaryotic expression vector of SLA-3 gene derived from Hebao and Laiwu pigs,which would lay a base for furter study the structure and function of the interest of proteins.

Key words: Hebao pig; Laiwu pig; SLA-3; expression

CLC Number:

GOU Dong-zhou, DAI Lin, YANG Jie, GAO Feng-shan. Construction and Expression of the Prokaryotic Expressing Vector of Pigs SLA-3 Derived from Two Chinese Local Strains[J]. , 2015, 42(2): 298-302.

References

[1] Willcox B E,Thomas L M,Bjorkman P J.Crystal structure of HLA-A2 bound to LIR-1,a host and viral major histocompatibility complex receptor[J].Nat Immunol,2003,4(9):913-919.
[2] O'Connor D H,Mothe B R,Weinfurter J T,et al.Major histocompatibility complex class Ⅰ alleles associated with slow simian immunodeficiency virus disease progression bind epitopes recognized by dominant acute-phase cytotoxic-T-lymphocyte responses[J].J Virol,2003,77(16):9029-9040.
[3] Bobe P,Benihoud K,Kiger N.Allogenic MHC class Ⅱ determinant(s) in MRL/Ipr autoimmune disease-prone mice.Unusual expression of an L1 transposable element creates molecular mimicry[J].J Immunol,1993,151(5):2813-2819.
[4] Hauptmann G,Bahram S.Genetics of the central MHC[J]. Curr Opin Immunol,2004,16(5):668-672.
[5] Trowsdale J,Campbell R D.Complexity in the major histocompatibility complex[J].Eur J Immunogenet,1992,19(1-2):45-55.
[6] Ohta Y,Shiina T,Lohr R L,et al.Primordial linkage of beta2-microglobulin to the MHC[J].J Immunol,2011,186(6):3563-3571.
[7] Ando A,Kawata H,Shigenari A,et al.Genetic polymorphism of the swine major histocompatibility complex (SLA) class Ⅰ genes,SLA-1,-2 and -3[J].Immunogenetics,2003,55(9):583-593.
[8] Lee J H,Simond D,Hawthorne W J,et al.Characterization of the swine major histocompatibility complex alleles at eight loci in Westran pigs[J].Xenotransplantation,2005,12(4):303-307.
[9] Lee Y J,Cho K H,Kim M J,et al.Sequence-based characterization of the eight SLA loci in Korean native pigs[J].Int J Immunogenet,2008,35(4-5):333-334.
[10] Ho C S,Martens G W,Amoss M S,et al.Swine leukocyte antigen (SLA) diversity in Sinclair and Hanford swine[J].Dev Comp Immunol,2010,34(3):250-257.
[11] Essler S E,Ertl W,Deutsch J,et al.Molecular characterization of swine leukocyte antigen gene diversity in purebred Pietrain pigs[J]. Anim Genet,2013,44(2):202-205.
[12] 姜巍,董宋鹏,李子彬,等.烟台黑猪SLA-2-YTH原核表达载体的构建及表达[J].中国畜牧兽医,2014,41(4):85-89.
[13] 高凤山,夏春,张强,等.大肠杆菌表达的重组猪β2微球蛋白二级结构的圆二色谱分析[J].微生物学报,2009,49(2):1596-1600.
[14] 陈茜茜,王京华,张香春,等.荷包猪SLA-2原核表达系的建立及表达研究[J].生物技术,2011,21(5):45-48.
[15] 白婧,李云岗,张强,等.六品系猪SLA-2多肽结合区分子结构差异研究[J].宁夏大学学报,2012,33(3):274-278.
[16] Kametani Y,Ohshima S,Kita Y F,et al.Porcine MHC classical class Ⅰ genes are coordinately expressed in superantigen-activated mononuclear cells[J].Vet Immunol Immunopathol,2012,148(3-4):252-259.
[17] Zhang N,Qi J,Feng S,et al.Crystal structure of swine major histocompatibility complex class Ⅰ SLA-1 0401 and identification of 2009 pandemic swine-origin influenza A H1N1 virus cytotoxic T lymphocyte epitope peptides[J]. J Virol,2011,85(22):11709-11724.
[18] Zhang J,Chen Y,Qi J,et al.Narrow groove and restricted anchors of MHC class Ⅰ molecule BF2 * 0401 plus peptide transporter restriction can explain disease susceptibility of B4 chickens[J].J Immunol,2012,189(9):4478-4487.
[19] 刘昌华,刘筏,高凤山,等.猪β2m四聚体前体链重组表达载体的构建和表达[J].中国畜牧兽医,2013,40(4):77-81.

Construction and Expression of the Prokaryotic Expressing Vector of Pigs SLA-3 Derived from Two Chinese Local Strains

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WEI Mingbang, BIANBA Qiongda, XIAO Qingqing, DUAN Mengqi, CHAMBA Yangzom, SHANG Peng. Cloning,Bioinformatics and Expression Analysis of CDKN1B Gene in Tibetan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2196-2206.
[2]	XIAO Peng, SHANG Jianghua, YANG Chunyan, LI Mengqi, DUAN Anqin, MA Xiaoya, FENG Chao, HUANG Chenqian, ZHANG Bo, ZHOU Jinchen, WEI Kelong, ZHENG Wei, ZHENG Haiying. Effects of Alpha-linolenic Acid on Buffalo Ovarian Granulosa Cells Cultured in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2380-2387.
[3]	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.
[4]	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.
[5]	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.
[6]	OU Yunwen, PAN Qin, WANG Yang, DAI Junfei, REN Shaoke, ZHANG Yang, ZHANG Jie. Truncated Expression of ORF2 Gene of Porcine Parvovirus Type 6 and Preparation of Its Polyclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2487-2495.
[7]	WANG Haorui, YU Baojun, YANG Jiayi, WEI Rong, FU Xi, WANG Chuanchuan, ZHANG Di, LI Desheng, CAI Zhengyun, GU Yaling, ZHANG Juan. Bioinformatics Analysis and Tissue Expression of DERA Gene in Jingyuan Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1764-1773.
[8]	PAN Pengcheng, LEI Zongquan, LI Xian, HU Xiangyun, QIN Qiantao, QIN Zhaoxian, GUAN Zhihui, CHEN Baojian, XIE Bingkun. Bioinformatics Analysis,Eukaryotic Expression Vector Construction and Tissue Expression of MYL2 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1796-1806.
[9]	ZHUO Na, ZHU Zhongwu, LI Jing, HU Shixiang, WANG Jianchang, LIN Hua, CHEN Chaolin, HAN Diangang, AI Jun, CHEN Peifu. Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1959-1970.
[10]	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.
[11]	WANG Xueping, ZHANG Xiaojun, LI Xiaoyue, WANG Guodong, ZHANG Fuliang, SONG Yuwei, ZHANG Mingliang, MA Lei. Prokaryotic Expression of Hexon Protein of Highly Pathogenic Fowl Adenovirus Serotype 4 and Preparation of Polyclonal Antibodies [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2053-2060.
[12]	TAO Qinghua, HUANG Caiyun, CHEN Chao, LIU Wei, SUN Ningning, LI Ang. Cloning and Tissue Expression Difference Analysis of LIF Gene in White Muscovy Duck [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1296-1306.
[13]	LIU Hongrun, ZHU Siran, FENG Lingli, ZHANG Kun, YAN Gang, WANG Yubin, ZHANG Shuai, JIANG Shan, XU Di, LAN Ganqiu, LIANG Jing. Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1352-1363.
[14]	ZHANG Zhewei, DAI Xiaotong, WEI Zudan, CHEN Yunpeng, GAN Jing, BEI Weicheng. Study on Prokaryotic Expression and Immunogenicity of OmpW and TbpA Proteins in Swine Pasteurella multocida [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1489-1498.
[15]	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.