非洲马瘟病毒VP7基因的克隆及其在昆虫细胞中的表达

›› 2012, Vol. 39 ›› Issue (1): 28-31.

• 生物技术 • Previous Articles Next Articles

Cloning and Expression of VP7 Gene of African Horse Sickness Virus in Baculovirus Infected Insect Cells

LI Fu-xiang¹, YANG Shi-biao¹, AI Jun², ZHOU Xiao-li², ZHAO Wen-hua¹, WANG Jin-ping¹

1. Yunnan Animal Science and Veterinary Institute, Kunming 650224,China;2. Yunnan Entry-Exit Inspection and Quarantine Bureau,Kunming 650228,China

Received:2011-06-21 Revised:1900-01-01 Online:2012-01-20 Published:2012-01-20

Abstract

Abstract: The VP7 gene fragment was cloned to the vector pFastBacHTB. The recombinant plasmid pFastBacHTB-AHSV-VP7 was constructed and was identified by PCR and enzyme digestion and sequenced. Then the plasmid pFastHTB-AHSV-VP7 was transformed into DH10Bac complement cells. It was identified by antibiotics and PCR, it showed the recombinant plasmid pFastBacHTB-AHSV-VP7 was constructed. On this basis, transposition bacmid DNA was extracted to transfect Sf9 insect cells.After transfected 96 hours, baculovirus was harvested.Then the Western blotting showed that the recombinant VP7 was expressed in insecet cells.The recombinant VP7 protein had a good biological activity and was approximate 45 ku. It is a base to construct a ELISA kit to test AHSV antibody and to prepare monoclonal antibody.

Key words: African horse sickness virus(AHSV); VP7 gene; baculovirus expression; insect cells

CLC Number:

S879.1

LI Fu-xiang;YANG Shi-biao;AI Jun;ZHOU Xiao-li;ZHAO Wen-hua;WANG Jin-ping. Cloning and Expression of VP7 Gene of African Horse Sickness Virus in Baculovirus Infected Insect Cells[J]. , 2012, 39(1): 28-31.

[1]	DUAN Xiangyuan, WU Jiangli, WEI Qiaohong, FENG Mao. Expression and Identification of Major Royal Jelly Protein 2 in Insect Cell-baculovirus System [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3779-3786.
[2]	LI Shuangxing, LIU Yebing, LIU Fangyuan, JI Jing, DU Jige, LI Qianlin, ZHU Zhen, LI Qihong, CHEN Xiaoyun, YIN Chunsheng. Expression and Identification of Feline Infectious Peritonitis Virus N Protein in Insect Cell-Baculovirus Expression System [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 273-279.
[3]	HE Haijian, WU Yuan, WANG Luyan, LI Qunjin, BA Shaobo, SHI Lin, SHAO Chunyan, SUN Jing, JIANG Sheng, WANG Xiaodu. Eukaryotic Expression of Recombinant Baculovirus of PEDV Isolated Strain S1 Gene [J]. , 2019, 46(3): 832-839.
[4]	CHEN Meirong, LIN Weidong, SONG Tianqi, XIN Ting, GUO Xiaoyu, HUANG Kehe, JIA Hong. Expression and Identification of H Gene of CDV BJ16B2 Strain in Insect Baculovirus Expression System [J]. , 2018, 45(8): 2041-2048.
[5]	WU Jing, XU Jian, HUANG Xiu-fen, SHEN Jun-jun, HE Hou-jun, LI Yong-qing. Expression and Biological Activity of Single Chain Antibody Against gD Glycoprotein of Infectious Bovine Rhinotracheitis Virus in Insect Cells [J]. , 2017, 44(8): 2475-2482.
[6]	WU Zhi, ZHANG Ji-ling, WU Meng, CUI Xiao-ting, WANG An-ping. Construction and Identification of Recombinant Avian Adeno-associated Virus Oviduct-specific Expressing Human Lysozyme Gene [J]. , 2017, 44(10): 2871-2877.
[7]	GUO Hui-juan, FAN Cui, ZHANG Ying, LV Mao-jie, LIANG Wu, YANG Bao-shou. Expression and Identification of Porcine Circovirus type 2 Capsid Protein Using flashBAC Baculovirus Expression System [J]. , 2016, 43(7): 1876-1883.
[8]	WANG An-ping, ZHU Shan-yuan, WANG Yong-juan, WU Shuang, ZUO Wei-yong, HONG Wei-ming. Expression and Identification of F Gene of Duck Newcastle Disease Virus in Insect Cells [J]. , 2015, 42(11): 2862-2866.
[9]	WANG Zhi-qiang，LI Hong-bin，YANG Xu-dong，HUANG Yu-xiang，ZOU Yue，ZHANG Jun，LIU Li-wei. Expression of Goose Parvovirus VP3 Gene in Baculovirus Expression System [J]. , 2014, 41(9): 52-55.
[10]	ZHAO Wen-nian, WANG Qing-hua, LIU Chun-ju, WANG Shu-juan, LEI Cheng-hong, BAO Jing-yue, WANG Zhi-liang. Expression and Identification of F Protein of Peste Des Petits Ruminants Virus Strain Tibet 07 in Insect Cells [J]. , 2014, 41(8): 29-33.
[11]	QUAN Chuan-song, JIANG Qian, YU Zuo, LI Bo-tao, LI Lian-feng, QU Lian-dong. Expression of Hemagglutinin of Canine Distemper Virus in the Insect Cell and its Antigenic Analysis [J]. , 2014, 41(8): 25-29.
[12]	LIU Chun-ju, REN Wei-jie, WANG Zhi-liang. Research Progress on Application of Baculovirus Expression System in Veterinary Subunit Vaccine Field [J]. , 2013, 40(9): 101-105.
[13]	JIN Li-ming, ZHANG Chun-lei, LIAO Ming, FAN Hui-ying. Construction and Identification of Recombinant Baculovirus Surface Displaying the Cap Protein of Porcine Circovirus Type 2 [J]. , 2013, 40(7): 6-10.
[14]	WU Chun-tao, LI Yan-mei. Cloning of gB, gE Gene of Bovine Herpesvirus-1 and Construction of its Baculovirus Expression Vector [J]. , 2012, 39(10): 73-75.
[15]	WANG Wen-xiu, SHEN Zhi-qiang, WANG Shan-hui, ZHANG Song-lin, CHI Xian-feng, XIA Xiao-jing. Expression of ORF2 Gene of Porcine Circovirus Type 2 in Sf9 Insect Cells [J]. , 2012, 39(10): 53-57.

Cloning and Expression of VP7 Gene of African Horse Sickness Virus in Baculovirus Infected Insect Cells

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments