Modification of the Full-length cDNA Clone of Newcastle Disease Virus of Goose Origin~* / 微生物学通报

ABSTRACT

The 6.5kb specific fragment containing the T7 promoter and the transcription vector was cut down from the full-length cDNA clone of Newcastle disease virus strain ZJI of goose origin,and thereafter it was self-ligated to form the high guality plasmid for mutagenesis.Site-directed mutagenesis technique was used for inserting three additional G nucleotides(nts) into the region between the T7 promter and the leader sequence of the NDV.The RT-PCR was employed to amplify the F/HN genes fragements,and then they were ligated by the shairing restriction enzyme BsmBI and finally the corresponding fragment in the mutatant full-length cDNA was substituted by 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 and all these studies lay a foundation for the research on the reverse genetics of NDV strain ZJI.