Modification of the full-length cDNA clone of Newcastle disease virus isolated from an outbreak in the goose
Modification of the full-length cDNA clone of Newcastle disease virus isolated from an outbreak in the goose
LIU Yuliang1, HU Shunli1, ZHANG Yanmei1, WU Yantao1, LIU Xiufan1, R?emer-Oberdoerfer Angela2, Veits Jutta2, Lange Martina2
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;
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.
出版日期: 2006-12-05
引用本文:
. Modification of the full-length cDNA clone of Newcastle disease virus isolated from an outbreak in the goose[J]. Frontiers of Biology in China, 2006, 1(4): 389-393.
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. Front. Biol., 2006, 1(4): 389-393.
