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 eDNA and a near oomplete genomic sequence that determine the granule-bound starch synthase {GBSS) gene from sago palm (Metroxylon sagu). The eDNA was isolated via a reverse transcription-polymerase chain reaction (RT -PCR) technique using primers designed bas...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2001
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/27918/2/isolation%20full.pdf |
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| Summary: | This study involved the isolation of a eDNA and a near oomplete genomic sequence that determine the
granule-bound starch synthase {GBSS) gene from sago palm (Metroxylon sagu). The eDNA 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 68-7:Z01o 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 confinned 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 ofthe sago GBSS via Southern hybridization. |
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