Begomoviruses: Occurrence and Management in Asia and Africa by Sangeeta Saxena & A. K. Tiwari

Author:Sangeeta Saxena & A. K. Tiwari
Language: eng
Format: epub
Publisher: Springer Singapore, Singapore

9.2 Mapping of Restriction Sites of Rose leaf curl virus and Its Betasatellite

Before the advent of the polymerase chain reaction (PCR), restriction enzyme digestion was the chief method for transferring the gene of interest from one DNA source to the other. For an instance, it was suggested that the potential of polymerase chain reaction to empower precise amplification of individual fragment of genome might restrict the utility of restriction enzymes. Nevertheless, restriction enzymes still have new efficacy acting as a diagnostic reagent to reveal that the DNA constructs have been ligated or placed in correct fashion. Restriction enzymes still attribute the inexpensive and the best suitable method to characterize DNA constructs (Roberts et al. 2003; Pingoud and Jeltsch 2001).

Restriction enzyme sets along with the magnitude of open reading frame were employed. The software exhibited the exact position of all the cut sites of restriction enzyme, and it also identifies the number of ORFs present in the geminivirus DNA. It subsequently demonstrates a schematic illustration of the geminivirus DNA comprising the ORFs sets which is laid on the guidelines of one cut only by all the restriction enzymes. The initial display also reveals that the enzymes can also be taken into consideration with a complete digestion to expunge every ORF (Table 9.1). Restriction enzymes further establish its use for characterizing the DNA of higher organisms via restriction fragment length polymorphisms (RFLPs) working as a physical marker or by directly identifying the occurrence of single nucleotide polymorphisms (SNPs). For analysing and constructing the restriction map (created by restriction enzymes by cleaving the DNA sequence) of the geminivirus DNA sequence, the software NEBcutter, version 1.0, was employed which is accessible online at http://​tools.​neb.​com/​NEBcutter and resulted in a diversity of displays (Vincze et al. 2003).Table 9.1Locations and the coding capability of the predicted ORFs for geminivirus DNA and the satellite molecule associated with it as extracted from the Rose plants

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