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| Construction of Recombinant CVI988 Virus Expressing GFP and Escherichia coli gpt Genes |
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Abstract Two fragments in the non-essential region of Marek's disease virus (MDV) were amplified by PCR from the genome of MDV CVI988 strain, and cloned into pUC19 for generating the 5.5 kb homologous recombinant arms. A transfer vector pUS-gpt-GFP was constructed by inserting the expressing cassettes of the CMV-gpt-ployA or CMV-GFP-polyA, respectively, into the unique BglⅡsite of US2 region in the recombinant arms. The transfer vector was transiently transfected into CHO cells and the expression of the green fluorescence protein was observed under the fluorescence microscope. This transfer vector was then transfected into CEF infected with MDV CVI988 strain. The recombinant CVI988 viruses expressing foreign genes, named rMDVgptGFP, were cloned by purifying the plagues expressing GFP in the MX-HAT selection medium. The purified recombinant viruses were further confirmed by PCR detection and growth of recombinant virus in selective and nonselective media.
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Received: 25 June 2008
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HAN Xue-Qing, GAO Feng-Ying, LU Mai-Xin, LIU Zhi-Gang, CAO Jian-Meng, WANG Miao, YI Meng-Meng, ZHANG De-Feng. Cloning, Expression and Functional Analysis of TRAF4 Gene in Nile Tilapia (Oreochromis niloticus)[J]. 农业生物技术学报, 2019, 27(3): 381-392. |
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