Construction of Recombinant Fowl Adenovirus Serotype 1 Exprsessing Exotic Green Fluorescence Protein

doi:10.3969/j.issn.1674-5817.2013.04.001

Abstract

Abstract: Objective To construct a recombinant fowl adenovirus expressing exotic green fluorescence protein(eGFP). Methods The flanking fragments of non-essential region(nt40 065~nt43 684) in fowl adenovirus genome were cloned into a transfer plasmid as well as eGFP. Through homologous recombination between the transfer plasmid and wild fowl adenovirus in chicken hepatocellular carcinoma cell line(LMH), the non-essential region between the flanking fragments was replaced by the complete eGFP expression cassette. In order to improve the occurrence probability of recombination, the transfection conditions such as logarithmic growth phase of LMH, the ratio of plasmid and lipofectamine, cells seeding density were explored. The optimum transfection condition was determined as follows: Seed 3×10⁵/ml LMH cells into 6 cell culture plate, 36 h later, cells were transfected with 8 μl lipofectamine 2 000 and 3.0 μg plasmid pUC-LR-eGFP. Recombinant virus was screening by plaque purification. Results recombinant adenovirus serotype 1 (rFAdV-1) expressing eGFP was obtained. Conclusion These results provided favorable references for fowl live vaccine development and related fundamental research.

Key words: Recombinant fowl adenovirus serotype 1(rFAdV-1), Chicken hepatocellular carcinoma cell line(LMH), Exotic green fluorescence protein(eGFP)

CLC Number:

Q95-33

ZHOU Jie, ZHAO Li, HU Jian-hua, GAO Cheng. Construction of Recombinant Fowl Adenovirus Serotype 1 Exprsessing Exotic Green Fluorescence Protein[J]. Laboratory Animal and Comparative Medicine, 2013, 33(4): 249-255.

References

[1] Logunov DY, Ilyinskaya GV, Cherenova LV, et al.Restoration of p53 tumor-suppressor activity in human tumor cells in vitro and in their xenografts in vivo by recombinant avain adenovirus CELO-p53[J]. Gene Therapy, 2004, 11:79-84.
[2] Shashkova EV, Cherenova LV, Kazansky DB, et al.Avain adenovirus vector CELO-TK displays anticancer activity in human cancer cells and suppresses established murine melanoma tumors[J]. Cancer Gene Therapy, 2005, 12:617-626.
[3] Francois A, Chevalier C, Delmas B, et al.Avain adenovirus CELO recombinants expressing VP2 of infectious bursal disease virus induce protection against bursal disease in chickens[J]. Vaccine, 2004, 22:2351-2360.
[4] Le Goff F, Méderlé-Mangeot I, Jestin A, et al.Deletion of open reading frame 9, 10 and 11 from the avain adenovirus CELO genome: effect on biodistribution and humoral responses[J]. J General Virol, 2005, 86:2019-2027.
[5] Rauw F, Lambrecht B, Francois A, et al.Kinetic and biologic properties of recombinant ChIFN-gamma expressed via CELO-virus vector[J]. J Interferon Cytokine Res, 2007, 27:111-118.
[6] 何秀苗. 禽腺病毒载体的构建及其生物学特性的研究[D]. 扬州: 扬州大学兽医学院, 2004.
[7] 王凤雪. CELO禽腺病毒载体的构建及fiber 1部分片段的原核表达[D]. 北京: 中国农业科学院, 2005.
[8] Tutykhina IL, Bezborodova OA, Shmarov MM, et al.Production of recombinant human lactoferrin in the allantoic fluid of embryonated chick eggs and its characteristics[J]. Protein Expr Purif, 2009, 65:100-107.
[9] Michou AI, Lehrmann H, Saltik M, et al.Mutational analysis of the avian adenovirus CELO, which provides a basis for delivery vectors[J]. J Virol, 1999, 73(2):1399-1410.
[10] Francois A, Eterradossi N, Delmas B, et al.Construction of avian adenovirus CELO recombinants in cosmids[J]. J Virol, 2001, 75(11):5288-5301.
[11] Cherenova LV, Logunov DY, Shashkova EV, et al.Recombinant avian adenovirus CELO expressing the human interleukin-2: characterization in vitro, in ovo and in vivo[J]. Virus Res, 2004, 100:257-261.

[1]	Ya ZHAO, Caiqin ZHANG, Han MENG, Jing QIN, Bing BAI, Yong ZHAO, Xu GE, Changhong SHI. Exploration of Laboratory Animal Science Teaching Practice from Perspectives of Curriculum Ideology and Politics [J]. Laboratory Animal and Comparative Medicine, 2023, 43(6): 641-646.
[2]	Xin LIU, Shaobo SHI, Cui ZHANG, Bo YANG, Chuan QU. Construction and Evaluation of End-to-side Anastomosis Model of Autologous Arteriovenous Fistula in Mice [J]. Laboratory Animal and Comparative Medicine, 2023, 43(6): 595-603.
[3]	Yongqiang NIE, Zhaoxia WANG. Rescue Technology and Its Application of Endangered Gene-Edited Mice [J]. Laboratory Animal and Comparative Medicine, 2023, 43(6): 636-640.
[4]	Yong ZHAO. Evolution and Prospects of Laboratory Animal Management: A Case Study of Shanghai's Development in the Past Decade [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 492-503.
[5]	Liping FENG, Qi ZHU, Jinxing LIN. Current Status and Reflection on the Study of Welfare for Laboratory Fish [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 524-530.
[6]	Jinxing LIN, Xindong WANG, Xuebing BAI, Liping FENG, Shuwu XIE, Qiusheng CHEN. Fine Structure of the Trunk Kidney and Distribution of Its Secreted Exosomes in the Adult Zebrafish [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 531-540.
[7]	Liya ZHAO, Liju NI, Caiqin ZHANG, Jianping TANG, Yangzheng YAO, Yanyan NIE, Xiaoxue GU, Ying ZHAO. Establishing a Genetic Detection Protocol of Single Nucleotide Polymorphisms Panels in Inbred Rats Based on Multiplex PCR-LDR [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 548-558.
[8]	Shuzhen ZHANG, Yanguang ZHAO. Investigation Report on the Production and Utilization Status of Experimental Mini-pigs [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 559-565.
[9]	Lingzhi YU, Jianyun XIE, Liping FENG, Xiaofeng WEI. Establishment of Fluorescence qPCR Method for Detection of Staphylococcus Aureus and Its Application in Feces Detection of Rats and Mice [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 566-573.
[10]	Chengji WANG, Jue WANG, Haijie WANG, Weisheng LU, Yan SHI, Zhengye GU, Mingqiu WAN, Ruling SHEN. Application of Optimized Latex Perfusion Technique in the Establishment of Craniofacial Venous Model in Mice [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 574-578.
[11]	Lianxiang GUO. Revision of Standards for Microbiological and Parasitological Grades in Laboratory Animals and Its Comparison to Foreign Standards [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 339-346.
[12]	Ying HUANG, Siyu WEI, Li CAI, Sujing QIANG, Dongting LI, Yuqiang DING. Microbiological Monitoring Analysis of Laboratory Rats and Mice from Vendors: Department of Laboratory Animal Science of Fudan University as an Example [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 347-354.
[13]	Xinyan BIAN, Yong LU, Yan WANG, Qiang SUN. Analysis of the Birthing Behaviour of Cynomolgus Macaques [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 355-362.
[14]	Yasheng DENG, Jiang LIN, Chiling GAN, Guanfeng ZENG, Jiayin HUANG, Huifang DENG, Yingxian MA, Siyin HAN. Literature Analysis of the Preparation Elements of Animal Models of Skin Photoaging and the Data of Subjects [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 406-414.
[15]	Hui CHENG, Fei FANG, Jiahao SHI, Hua YANG, Mengjie ZHANG, Ping YANG, Jian FEI. H1 Linker Histone Gene Regulates Lifespan via Dietary Restriction Pathways in Caenorhabditis elegans [J]. Laboratory Animal and Comparative Medicine, 2023, 43(3): 271-281.