Preparation of GFLV CP-gene cDNA probe labeled by photosensitive biotin and its application in detection of GFLV.
The cDNA of the GFLV CP gene amplified from clone pGAB5 by PCR was labeled by photosensitive biotin. The probe sensitivity is 5 pg, and the detectable amount of unlabeled GFLV cDNA was 10 pg by Dot-blot hybridization when the biotin-labeled cDNA was used as a probe. The results of detection for GFLV by probe have positive correlation with those of ELISA. The minimum amount of grapevine tissue for detection by probe was 100 times lower than that by ELISA. The seasons and the position of grapevine leaves do not limit application of this method.
Cao
X
X
Laboratory of Plant Biotechnology, Chinese Academy of Sciences, Beijing, China.
Zhang
Y
Y
Cai
W
W
Guan
W
W
Mang
K
K
D019921:Biotinylation; D002213:Capsid / Q000032:analysis / Q000235:genetics*; D036022:Capsid Proteins; D015342:DNA Probes / Q000235:genetics*; D018076:DNA, Complementary; D004797:Enzyme-Linked Immunosorbent Assay; D015336:Molecular Probe Techniques; D018515:Plant Leaves / Q000737:chemistry / Q000821:virology; D010942:Plant Viruses / Q000235:genetics*; D016133:Polymerase Chain Reaction; D019685:Rosales / Q000821:virology*
D036022:Capsid Proteins; D015342:DNA Probes; D018076:DNA, Complementary; C095208:coat protein, grapevine fanleaf virus