Modification of the full-length cDNA clone of Newcastle disease virus isolated from an outbreak in the goose

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  • 1.Key Laboratory of Animal Infectious Diseases, Ministry of Agriculture, Yangzhou University, Yangzhou 225009, China; 2.Institute of Molecular Biology, Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, D-17498 Insel Riems, Germany;

Published date: 05 Dec 2006

Abstract

A 6.5-kb specific fragment containing the T7 promoter and the transcription vector was excised from the full-length cDNA clone of the Newcastle disease virus (NDV) strain ZJI of goose origin, and thereafter it was self-ligated to form a high quality plasmid for mutagenesis. Site-directed mutagenesis was used for inserting three additional G nucleotides (nts) into the region between the T7 promoter and the leader sequence of the NDV genome. RT-PCR was employed to amplify the F/HN gene fragments, and then they were ligated by the shared restriction enzyme BsmBI. Finally, the corresponding fragment in the mutant full-length cDNA was substituted with the new one. The sequencing results showed that the three additional G nts were successfully inserted and the mutant nts in the full-length cDNA were corrected. This study lays a good foundation for research on the reverse genetics of NDV strain ZJI.

Cite this article

LIU Yuliang, HU Shunli, ZHANG Yanmei, WU Yantao, LIU Xiufan, R?emer-Oberdoerfer Angela, Veits Jutta, Lange Martina . Modification of the full-length cDNA clone of Newcastle disease virus isolated from an outbreak in the goose[J]. Frontiers in Biology, 2006 , 1(4) : 389 -393 . DOI: 10.1007/s11515-006-0051-6

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