id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-296364-7rp60d2m	Youn, Soonjeon	In vitro assembled, recombinant infectious bronchitis viruses demonstrate that the 5a open reading frame is not essential for replication	2005-02-05		.txt	text/plain	5112	270	56	Molecular clones of infectious bronchitis virus (IBV), derived from the Vero cell adapted Beaudette strain, were constructed, using an in vitro assembly method. Direct sequencing of RT-PCR products derived from cells infected with the plaque-purified virus, which had lost expression of EGFP, confirmed loss of the EGFP ORF. This strategy was further modified to construct an infectious cDNA clone of MHV with their bno see'mQ technology, in which restriction endonuclease sequences were incorporated into amplicons, such that upon enzyme treatment, the endonuclease sites were eliminated prior to in vitro ligation. In the current study, molecular clones of IBV were generated using the in vitro assembly of cDNA fragments as a template for transcription of full-length genomic RNA. Interestingly, the size of plaques with the 5a deletions, including a revertant that had completely lost the EGFP ORF, was comparable to plaques generated by our standard Beaudette cell-adapted Beaudette virus (Fig. 6B ).	./cache/cord-296364-7rp60d2m.txt	./txt/cord-296364-7rp60d2m.txt