TI: |
Title
Integration and expression of Bluetongue VP2 gene in somatic embryos of peanut through particle bombardment method |
AU: |
Author
Athmaram, TN; Bali, Geetha; Devaiah, KM |
AF: |
Author Affiliation
Department of Microbiology and Biotechnology, Bangalore University, Jnanabharathi, Bangalore 560056, India, baliccet@vsnl.com |
SO: |
Source
Vaccine [Vaccine]. Vol. 24, no. 15, pp. 2994-3000. 2006. |
IS: |
ISSN
0264-410X |
DE: |
Descriptors
Kanamycin; Somatic embryos; Transgenic plants; Nuts; Plantlets; Polymerase chain reaction; VP2 protein; Epitopes; particle bombardment; Reverse transcription; Plasmids; Osmosis; Vaccines; Somatic embryogenesis; Gene transfer; Callus; Coat protein; DNA probes; Bluetongue; Integration; Enzymes; Arachis hypogaea; Bluetongue virus |
AB: |
Abstract
After pre-culture and treatment of osmosis, zygotic embryos of peanut (Arachis hypogaea L.) were transformed via particle bombardment with a plasmid containing a Bluetongue VP2 gene (BTVP2) comprising neutralizing epitopes. Selection for Kanamycin resistant calluses and somatic embryos was initiated at 12th day post-bombardment on medium containing 25 mg/L Kanamycin. Under continuous selection, 12.38% Kanamycin resistant plantlets were regenerated from bombarded somatic embryos. The presence and integration of BTVP2 DNA in regenerated Kanamycin resistant plants were confirmed by southern hybridization assay using non-radioactive Digoxiginin BTVP2 probe. beta - Glucuronidase (GUS) enzyme activity was detected in transgenic somatic embryos but not from control, non-transformed embryos. The expression of the BTVP2 protein was confirmed through RT-PCR (reverse transcription polymerase chain reaction) using the RNA isolated from the transgenic callus employing BTVP2- specific primers. The production of transgenic peanut was mainly focused on evaluating a newly improved somatic embryogenesis regeneration system as well as the gene transfer method and to produce the Bluetongue outer coat protein that comprises the neutralizing epitopes. |
LA: |
Language
English |
SL: |
Summary Language
English |
PY: |
Publication Year
2006 |
PT: |
Publication Type
Journal Article |
PB: |
Publisher
Elsevier Science Ltd., The Boulevard Langford Lane Kidlington
Oxford OX5 1GB UK, nlinfo- f@elsevier.nl, http://www.elsevier.nl |
DO: |
DOI
10.1016/j.vaccine.2005.11.019 |
CL: |
Classification
W2 32065 Plants; V 22098 Immunization: Vaccines & vaccination: Animal; F 06700 Monoclonal antibodies, hybridoma, antigens |
UD: |
Update
200604 |
SF: |
Subfile
Virology & AIDS Abstracts; Agricultural and Environmental Biotechnology Abstracts; Immunology Abstracts |
AN: |
Accession Number
6719706 |
PG: |
Journal Pages
2994-3000 |
JV: |
Journal Volume
24 |
JI: |
Journal Issue
15 |