Isolation Of Cdna Fragment Encoding Starch Synthase Gene From Metroxylon Sagu By Rt Pcr Method

Isolation of Cdna Fragment Encoding Starch Synthase Gene from Metroxylon Sagu by Rt-pcr Method

Isolation of CDNA Fragment Encoding Starch Branching Enzyme (Isoform) Gene from Metroxylon Sagu by Rt Pcr Method

Isolation and Characterization of Near Complete CDNA and Genomic Sequences Coding for Granule-bound Starch Synthase in Sago Palm (Metroxylon Sagu)

This study involved the isolation of a cDNA and a near complete genomic sequence that determine the granule-bound starch synthase (GBSS) gene from sago palm (Metroxylon, sagu). The cDNA was isolated via a reverse transcription-polymerase chain reaction (RT-PCR) technique using primers designed based on the conserved regions of the GBSS. The RT-PCR product obtained, 917 bp in size, was cloned and sequenced. Homology search by using the BLASTX search program confirmed that the RT-PCR product consists of part of the GBSS gene, with the deduced amino acid sequence sharing 68-72% identities with GBSS of rice, potato, maize, wheat and cassava. A pair of specific internal primers was then designed based on the RT-PCR product. These primers were used to amplify the corresponding genomic DNA region via a genomic PCR method. A 2,008 bp genomic PCR product was generated by this primer set. Cloning and sequencing of the genomic PCR product confirmed that it consists of a partial genomic GBSS gene. This genomic PCR product was then used as a probe in an attempt to isolate the entire genomic locus of sago GBSS via Southern hybridization.