TY - JOUR
T1 - Transgenic apple expressing an antigenic protein of the human respiratory syncytial virus
AU - Lau, Joann M.
AU - Korban, Schuyler S.
N1 - Funding Information:
This work was funded by a USDA-IFAFS grant. The authors would like to thank Dr. Sergei Krasnyanski for his assistance with apple transformation and Ms. Elise Bendik for her assistance with ELISA assays. We also like to acknowledge Dr. Jagdeep Sandhu and Dr. Dennis Buetow for their earlier work on developing the constructs used in this study.
PY - 2010/7/15
Y1 - 2010/7/15
N2 - A gene coding for the human respiratory syncytial virus (RSV)-F protein, driven by the constitutively expressed CaMV 35S promoter, was introduced into leaf tissues of apple, Malus×domestica Borkh. cv. Royal Gala, via Agrobacterium-mediated transformation. Two putative transgenic lines were identified, and the presence of the RSV-F gene was confirmed by polymerase chain reaction (PCR). A total of 25 plants from these different transgenic events were successfully rooted, acclimatized, and transferred to the greenhouse. Stable integration of the transgene was confirmed and transgene copy number was determined by DNA gel blot analysis. Expression of the npt-II selectable marker and RSV-F was determined using reverse-transcription polymerase chain reaction (RT-PCR). Furthermore, enzyme-linked immunosorbent assay (ELISA) revealed varying levels of protein expression of the RSV-F transgene, ranging from 0 to 20 μg/g tissue. This is a first step in an effort to assess the efficacy of using apple for developing a plant-based vaccine against RSV.
AB - A gene coding for the human respiratory syncytial virus (RSV)-F protein, driven by the constitutively expressed CaMV 35S promoter, was introduced into leaf tissues of apple, Malus×domestica Borkh. cv. Royal Gala, via Agrobacterium-mediated transformation. Two putative transgenic lines were identified, and the presence of the RSV-F gene was confirmed by polymerase chain reaction (PCR). A total of 25 plants from these different transgenic events were successfully rooted, acclimatized, and transferred to the greenhouse. Stable integration of the transgene was confirmed and transgene copy number was determined by DNA gel blot analysis. Expression of the npt-II selectable marker and RSV-F was determined using reverse-transcription polymerase chain reaction (RT-PCR). Furthermore, enzyme-linked immunosorbent assay (ELISA) revealed varying levels of protein expression of the RSV-F transgene, ranging from 0 to 20 μg/g tissue. This is a first step in an effort to assess the efficacy of using apple for developing a plant-based vaccine against RSV.
KW - Agrobacterium-mediated transformation
KW - Malus×domestica
KW - Plant-based vaccine
KW - Respiratory syncytial virus
KW - Transgene expression
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U2 - 10.1016/j.jplph.2010.02.003
DO - 10.1016/j.jplph.2010.02.003
M3 - Article
C2 - 20307914
AN - SCOPUS:77953132273
SN - 0176-1617
VL - 167
SP - 920
EP - 927
JO - Journal of Plant Physiology
JF - Journal of Plant Physiology
IS - 11
ER -