scholarly journals GENETIC ENGINEERING OF THE ZUCCHINI YELLOW MOSAIC VIRUS COAT PROTEIN GENE FOR EXPRESSION IN PLANTS.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1130d-1130
Author(s):  
Guowei Fang ◽  
Rebecca Grumet
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Zucchini Yellow Mosaic Virus ◽  
In Vitro Transcription ◽  
Yellow Mosaic Virus ◽  
35S Promoter ◽  
Plant Expression Vector

Zucchini yellow mosaic virus (ZYMV), a potyvirus, can cause major losses in cucurbit crops. With the goal of genetically engineering resistance to this disease we have engineered the ZYMV coat protein gene into a plant expression vector. The complete coat protein coding sequence, or the conserved core portion of the capsid gene, was attached to the 5' untranslated region of tobacco etch virus (TEV) in the pTL37 vector (Carrington et al., 1987, Nucl. Acid Res. 15:10066) The capsid constructs were successfully expressed by in vitro transcription and translation systems as verified by SDS-PAGE and ZYMV coat protein antibody. The constructs were then subcloned using polymerase chain reaction and attached to the CaMV 35 S transcriptional promoter on the CIBA-GEIGY pCIB710 plasmid. The constructs containing the CaMV 35S promoter, the 5' untranslated leader of TEV, and ZYMV coat protein sequences were then put between the Agrobacterium tumefaciens left and right borders in the pCIB10 vector and transferred to A. tumefaciens strain LBA4404 by triparental mating. These vectors are now being used to transform muskmelon and cucumber; resultant transgenic plants will be tested for ZYMV coat protein expression.

scholarly journals Isolation and Molecular Characterization of a Chilean Isolate of Zucchini yellow mosaic virus

Plant Disease ◽  
2001 ◽  
Vol 85 (6) ◽  
pp. 644-648 ◽  
Author(s):  
H. Prieto ◽  
A. Bruna ◽  
P. Hinrichsen ◽  
C. Muñoz
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sequence Comparisons ◽  
Severe Stunting ◽  
Chilean Isolate

Zucchini yellow mosaic virus (ZYMV) was described in 1981 affecting squash, melon, and other cultivated cucurbits with severe stunting and yellowing symptoms. It was reported to be present in most countries where cucurbits are grown, and in Chile since 1995, from surveys using enzyme-linked immunosorbent assay (ELISA) but without further characterization. A potyvirus was isolated from ELISA-positive symptomatic plants. The results indicate that this virus is ZYMV based on symptoms on herbaceous indicators, immunospecific electron microscopy of the purified particle, and sequencing of 395 bases of the 3′ end of the coat protein gene. The virus was detected in melon, watermelon, and squash plants. In agreement with previous descriptions for ZYMV, the Chilean isolate is a flexuous filamentous particle 740 nm long with one main protein of approximately 36 kDa. Nucleotide sequence comparisons of the 3′ portion of the coat protein gene revealed a high similarity to the Connecticut and California strains.

The sense and antisense coat protein gene of alfalfa mosaic virus strain N20 confers protection in transgenic tobacco plants

1997 ◽  
Vol 48 (4) ◽  
pp. 503 ◽  
Author(s):  
K. W. Jayasena ◽  
B. J. Ingham ◽  
M. R. Hajimorad ◽  
J. W. Randles
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Alfalfa Mosaic Virus ◽  
35S Promoter ◽  
Pasture Legumes ◽  
Nicotiana Tabacum L ◽  
Sense Orientation ◽  
A Minor

The coat protein gene of a South Australian strain of alfalfa mosaic virus (AMV-N20 [NcS]) has been cloned, sequenced, and transferred into Nicotiana tabacum L. cv. Xanthi via Agrobacterium tumefaciens under the control of the CaMV 35S promoter. A number of lines (T0 generation) were selected with the coat protein gene either in sense orientation (CP+) or in antisense orientation (CP–). The T0 plants were tested for their gene expression and susceptibility to the homologous AMV strain. A significant delay in the onset of symptoms and a reduction in virus accumulation was observed in CP+ plants mechanically inoculated with AMV. CP– plants were also significantly protected but less so than the CP+ plants. Plants transformed with the expression vector only (CP0) showed a minor resistance to local infection on inoculated leaves compared with untransformed plants. The strategy of coat protein mediated protection (CPMP) using the CP gene in either messenger sense or antisense would therefore be appropriate for testing on economically important pasture legumes.

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