Primer Design for Isolation of Sucrose:Sucrose 1-Fructosyltransferase (1-SST) Gene from Gembili (Dioscorea esculenta)

Recent studies have reported the presence of inulin, a prebiotic polysaccharide, in gembili (Dioscorea esculenta). Sucrose:sucrose 1-fructosyltransferase (1-SST) is an enzyme that catalyzes the first step of inulin biosynthesis. The identification of this enzyme would be the foundation to improve the yield of inulin in gembili; to modify 1-SST gene for this purpose, its sequence must first be determined. This study aimed to design primers to isolate 1-SST gene from D. esculenta. The primers were designed by using the whole-genome sequence of Dioscorea rotundata due to the lack of genomic information on D. esculenta. Sequences from chromosome 6 and 11 were used as the template of primer design in which four pairs of primers were selected. Amplification products with expected size were gel-purified, then the targets were sequenced and analyzed in-silico. As a result, one of the primer pairs had successfully isolated vacuolar invertase gene, which is closely related to 1-SST gene. On the other hand, the other primer pairs showed either negative or false-positive result. Using the current strategy, 1-SST gene sequence from D. esculenta has not been successfully isolated, thus other approaches in primer design should be considered in further studies. Keywords: 1-SST, Dioscorea esculenta, Gembili, Inulin, Primer desig

Primer Design for Isolation of Sucrose:Sucrose 1-Fructosyltransferase (1-SST) Gene From Gembili (Dioscorea Esculenta)

Recent studies have reported the presence of inulin, a prebiotic polysaccharide, in gembili (Dioscorea esculenta). Sucrose:sucrose 1-fructosyltransferase (1-SST) is an enzyme that catalyzes the first step of inulin biosynthesis. The identification of this enzyme would be the foundation to improve the yield of inulin in gembili; to modify 1-SST gene for this purpose, its sequence must first be determined. This study aimed to design primers to isolate 1-SST gene from D. esculenta. The primers were designed by using the whole-genome sequence of Dioscorea rotundata due to the lack of genomic information on D. esculenta. Sequences from chromosome 6 and 11 were used as the template of primer design in which four pairs of primers were selected. Amplification products with expected size were gel-purified, then the targets were sequenced and analyzed in-silico. As a result, one of the primer pairs had successfully isolated vacuolar invertase gene, which is closely related to 1-SST gene. On the other hand, the other primer pairs showed either negative or false-positive result. Using the current strategy, 1-SST gene sequence from D. esculenta has not been successfully isolated, thus other approaches in primer design should be considered in further studies. Keywords: 1-SST, Dioscorea esculenta, Gembili, Inulin, Primer desig