含非洲马瘟病毒部分核酸序列的病毒样颗粒的研制

doi:10.11843/j.issn.0366-6964.2015.04.018

Abstract

Abstract:

This experiment was conducted to prepare the virus-like particles containing the partial gene fragment of African horse sickness virus (AHSV).Genome segment 7 (S7) encoding protein VP7，the group-specific protein of AHSV，is a primary target for reverse transcription-PCR and real time reverse transcription-PCR detection of AHSV.The conservative sequence of S7 gene was synthesized and cloned into plasmid pNH-MS2his，which contained the coat protein gene of E.coli bacteriophage MS2 and His tag，to construct the prokaryotic express vector pNH-MS2his-VP7.The recombinant plasmid pNH-MS2his-VP7 was transformed into BL21 (DE3).Expression of the target protein was induced by IPTG for 5 h.The virus-like particles containing the S7 RNA sequence of AHSV were simply obtained by induction and sequent Ni+ chromatography purification.The virus-like particles were confirmed have good uniformity and stability.They have great prospect as the standard and quality control in the detection of AHSV.

Key words: African horse sickness virus, His tag, virus-like particle, detection

CLC Number:

S852.659.4

SUN Min，LIANG Cheng-zhu，WANG Qun，ZHANG Xiao-wen，XIAO Xi-zhi，GENG Juan，CHEN Ji-xiang. Preparation of the Virus-like Particles Containing the Partial Gene Fragment of AHSV[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(4): 637-643.

References

［1］世界动物卫生组织.陆生动物诊断试验和疫苗手册［M］.2007.
World Organisation for Animal Health.Manual of diagnostic tests and vaccines for terrestrial animals［M］.2007.(in Chinese)
［2］赵文华，杨仕标，王金萍，等.非洲马瘟病毒群特异性RT-PCR检测方法的研究［J］.生物技术通报，2011（4）：204-207.
ZHAO W H，YANG S B，WANG J P，et al.Development of RT-PCR method for detection of AHSV［J］.Biotechnology Bulletin，2011（4）：204-207.(in Chinese)
［3］曾昭文，花群义，段纲.非洲马瘟病毒分子生物学研究进展［J］.动物医学进展，2006，27（10）：1-6.
ZENG Z W，HUA Q Y，DUAN G.Advance in molecular biology of African Horse Sickness Virus［J］.Progress in Veterinary Medicine，2006，27（10）：1-6.(in Chinese)
［4］高志强，张鹤晓，赖平安，等.非洲马瘟病毒VP7基因拼接、表达及重组ELISA方法的建立与初步应用［J］.畜牧兽医学报，2008，39（11）：1548-1553.
GAO Z Q，ZHANG H X，LAI P A，et al.Development and application of the recombinant ELISA based on the assembling and expression of VP7 gene of African horse sickness virus［J］.Acta Veterinaria et Zootechnica Sinica，2008，39（11）：1548-1553.(in Chinese)
［5］张海明，段晓冬，相文华，等.非洲马瘟诊断技术研究概况［J］.动物医学进展，2013，34（6）：175-178.
ZHANG H M，DUAN X D，XIANG W H，et al.Progress on diagnostic techniques for African Horse Sickness［J］. Progress in Veterinary Medicine，2013，34（6）：175-178.(in Chinese)
［6］中国计量科学研究院.JJF 1343—2012标准物质定值的通用原则及统计学原理［S］.北京：中国标准出版社，2012.
National Institute of Metrology.JJF 1343-2012 general and statistical principles for characterization of reference materials［S］.Beijing：Standards Press of China，2012.(in Chinese)
［7］ZHAN S，LI J M，XU R H，et al.Armored long RNA controls or standards for branched DNA assay for detection of human immunodeficiency virus type 1［J］.J Clin Microbiol，2009，47(8)：2571-2576.
［8］STEVENSON J，HYMAS W，HILLYARD D.The use of armored RNA as a multi-purpose internal control for RT-PCR ［J］. J Virol Methods，2008，150（1-2）：73-76.
［9］CHENG Y J，NIU J J，ZHANG Y Y，et al.Preparation of His-tagged armored RNA phage particles as a control for real-time reverse transcription-PCR detection of severe acute respiratory syndrome coronavirus ［J］. J Clin Microbiol，2006，44(10)：3557-3561.
［10］WEI Y X，YANG C M，WEI B J，et al.RNase-resistant virus-like particles containing long chimeric RNA sequences produced by two-plasmid coexpression system ［J］.J Clin Microbiol，2008，46(5)：1734-1740.
［11］窦敏，张国广，于广福，等.含口蹄疫病毒IRES RNA病毒样颗粒表达载体的构建［J］.中国生物工程杂志，2007，27（9）：31-35.
DOU M，ZHANG G G，YU G F，et al.Construction of the expression vector of virus-like particles containing FMDV IRES RNA［J］.China Biotechnology，2007，27（9）：31-35.(in Chinese)
［12］彭建明，李金明，徐克前，等.含人甲胎蛋白mRNA部分序列的耐核糖核酸酶病毒样颗粒的构建和表达［J］.中华肝脏病杂志，2005，13(4)：304-306.
PENG J M，LI J M，XU K Q，et al.Construction and expression of RNase-resisting virus-like particles containing partial sequence of alpha-fetoprotein messenger RNA［J］.Chinese Journal of Hepatology，2005，13(4)：304-306.(in Chinese)

[1]	WANG Suhua, SHUAI Jiangbing, LI Zhou, YUAN Shuhui, WU Shaoqiang, Lü Jizhou, ZHAO Zhiguo. Establishment and Application of a Duplex Real-time PCR Assay for Detection of Bacillus anthracis [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(8): 1658-1665.
[2]	WANG Suhua, SHUAI Jiangbing, LI Zhou, YUAN Shuhui, WU Shaoqiang, Lü Jizhou, ZHAO Zhiguo. Establishment and Application of a Duplex Real-time PCR Assay for Detection of Bacillus anthracis [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(8): 1658-1665.
[3]	YU Yan, GU Yu, WANG Chengdong, HU Jing, YANG Aining, YANG Qiao, LI Desheng, ZHANG Hemin, DENG Linhua, LING Shanshan, ZUO Zhicai, PENG Guangneng, ZHONG Zhijun, MA Xiaoping. Dynamic Changes of Dectin-1, TLR-2,TLR-4 and Cytokines in Mice Infected with Microsporum gypseum [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1449-1457.
[4]	YU Yan, GU Yu, WANG Chengdong, HU Jing, YANG Aining, YANG Qiao, LI Desheng, ZHANG Hemin, DENG Linhua, LING Shanshan, ZUO Zhicai, PENG Guangneng, ZHONG Zhijun, MA Xiaoping. Dynamic Changes of Dectin-1, TLR-2,TLR-4 and Cytokines in Mice Infected with Microsporum gypseum [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1449-1457.
[5]	ZHENG Lanlan, ZHU Jingjing, WANG Pan, SHU Yan, LIANG Qingqing, LI Bingxiao, WANG Chaoqun, WEI Zhanyong. Development and Application of a TaqMan based Real-time Fluorescent RT-PCR for Specific Detection of Porcine Deltacoronavirus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1261-1267.
[6]	QIN Tong, ZHOU Ling, YOU Xinyue, LIANG Lin, SHI Lijun, ZHANG Jianwei, LI Jinxiang, CUI Shangjin. Establishment and Application of Nano PCR Assay for Detection of the Canine Parvovirus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1268-1274.
[7]	FAN Hui, LI Liang, JIANG Ping, WANG Xianwei, LI Yufeng, BAI Juan. Establishment and Application of a RT-PCR Assay for Detection of Senecavirus A(SVA) [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1312-1318.
[8]	ZHENG Lanlan, ZHU Jingjing, WANG Pan, SHU Yan, LIANG Qingqing, LI Bingxiao, WANG Chaoqun, WEI Zhanyong. Development and Application of a TaqMan based Real-time Fluorescent RT-PCR for Specific Detection of Porcine Deltacoronavirus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1261-1267.
[9]	QIN Tong, ZHOU Ling, YOU Xinyue, LIANG Lin, SHI Lijun, ZHANG Jianwei, LI Jinxiang, CUI Shangjin. Establishment and Application of Nano PCR Assay for Detection of the Canine Parvovirus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1268-1274.
[10]	FAN Hui, LI Liang, JIANG Ping, WANG Xianwei, LI Yufeng, BAI Juan. Establishment and Application of a RT-PCR Assay for Detection of Senecavirus A(SVA) [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1312-1318.
[11]	GUO Zijing, HE Qifu, TANG Cheng, ZHANG Bin, YUE Hua. Establishment and Application of a Real-time RT-PCR Assay for Detecting Bovine Nebovirus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 893-900.
[12]	GUO Zijing, HE Qifu, TANG Cheng, ZHANG Bin, YUE Hua. Establishment and Application of a Real-time RT-PCR Assay for Detecting Bovine Nebovirus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 893-900.
[13]	WANG Junwei, CHEN Fangzhou, KU Xugang, LI Chang, HE Qigai. Establishment and Clinical Application of an Indirect ELISA for Detection of Antibodies against PCV3 Based on Recombinant ORF2 Protein [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 454-460.
[14]	WANG Junwei, CHEN Fangzhou, KU Xugang, LI Chang, HE Qigai. Establishment and Clinical Application of an Indirect ELISA for Detection of Antibodies against PCV3 Based on Recombinant ORF2 Protein [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 454-460.
[15]	LU Zengkui, ZHANG Liping, LI Qing, LIU Enmin, DU Lixin, CHU Mingxing, WEI Caihong. N⁶-methyladenosine Modification in mRNA and Its Research Advance in Animals [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(1): 1-13.

Preparation of the Virus-like Particles Containing the Partial Gene Fragment of AHSV

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments