full issue - Association of Biotechnology and Pharmacy

Current Trends in Biotechnology and PharmacyVol. 5 (2) 1163-1172 April 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1167a)b)Fig. 3. a) GQ496588. Coscinium fenestratum internal transcribed spacer 1, complete sequence; and 5.8Sribosomal RNA gene, partial sequence b) GQ496589.Coscinium fenestratum 5.8S ribosomal RNA gene, partialsequence; and internal transcribed spacer 2, complete sequenceand their homology percentage with the respectiveITS region of C. fenestratum is given in the Table3. Variations in the ITS sequences was observedamong the different species studied.Development and Validation of Speciesspecificmarker for C. fenestratum: NCBIprimer blast tool was used to develop and validatethe species-specific markers for C. fenestratum.On in silico analysis, a primer set (CF1F andCF1R) indicated a high specificity foramplification of 196 bp only from the ITS1sequence of C. fenestratum (Table 4). Thesynthesized oligos were incorporated in a PCRreaction individually with the genomic DNA ofC. fenestratum and its adulterant species.Electrophoretic separation of the ampliconindicated 196 bp product only with the C.fenestratum samples and it did not give any nonspecificamplification with the other test speciesunder study (Figure 4).DiscussionHerbal medicinal products may vary incomposition and properties, unlike conventionalpharmaceutical products prepared from synthetic,chemically pure materials by means ofFig. 4. PCR amplification with CF1F and CF1R using the genomic DNA of C. fenestratum and its adulterant species LaneM – 100 bp ladder; 1 to 5 – C. fenestratum accessions; Lane 6 to 8 – B. aristata; Lane 9 to 11 – B. asiatica; Lane 12 to 14– B. lycium; Lane 15 to 17 – M. umbellataMolecular Marker to distinguish Coscinium fenestratum