請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/68306
標題: Construction of in vitro and in vivo infectious transcripts of a Taiwan strain of Zucchini yellow mosaic virus
作者: Lin, S.S.
Hou, R.F.
Yeh, S.D.
關鍵字: in vitro
in vivo
infectious transcripts
ZYMV TW-TN3
plum pox potyvirus
vein mottling virus
length cdna clone
particle
bombardment
in-vitro
invitro transcripts
beta-glucuronidase
intron-insertion
escherichia-coli
tobacco
期刊/報告no:: Botanical Bulletin of Academia Sinica, Volume 43, Issue 4, Page(s) 261-269.
摘要: The full-length cDNA of a Taiwan strain of Zucchini yellow mosaic virus (ZYMV TW-TN3) was constructed from five overlapping cDNA clones downstream from the bacteriophage T7 promoter in plasmid pT7ZYMV2-5. The plasmid was able to generate an in vitro transcript corresponding to TW-TN3 (9591 nt) with one extra guanosine residue at the 5' terminus and a poly(A)(95) tract at the 3' end. In addition, pT7ZYMV2-5 was used for the construction of p35SZYMV2-26 that contained the full-length cDNA of TW-TN3 with a Cauliflower mosaic virus (CaMV) 35S promoter and a nopaline synthase (nos) terminator. The capped in vitro transcript generated from pT7ZYMV2-5 and the purified DNA of p35SZYMV2-26 were introduced into zucchini squash plants by mechanical inoculation and particle bombardment, respectively. Both in vitro and in vivo transcripts induced systemic symptoms on zucchini squash 4 to 6 days after inoculation. In addition, both transcripts also induced local lesions on plants of Chenopodium quinoa by mechanical inoculation. The results of infectivity assay, symptomatology, and serologically specific electron microscopy indicated that the in vitro and in vivo TW-TN3 transcripts derived from pT7ZYMV2-5 and p35SZYMV2-26, respectively, are infectious. The ability to generate biologically functional transcripts from the constructed cDNA clones is a significant step for molecular analyses of TW-TN3.
URI: http://hdl.handle.net/11455/68306
ISSN: 0006-8063
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