Cloning and characterization of putative pcr-amplified adp-glucose pyrophosphorylase (AGPase) small subunit gene in Sago

Toh, Janice Pei Yin (2006) Cloning and characterization of putative pcr-amplified adp-glucose pyrophosphorylase (AGPase) small subunit gene in Sago. [Final Year Project Report] (Unpublished)

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Abstract

Isolation of small subunit ADP-glucose pyrophosphorylase (AGPase) gene from sago palm (Metrorylon sagu) was perfonned. Two sets of primers, namely haAGP and ha2AGP, were used in this study and both primers yielded PCR products. From all the multiple PCR bands generated by both of primers, only two PCR fragments generated by primer set haAGP and one PCR fragment amplified using ha2AGP were used for subsequent study. The sizes of the PCR fragment selected were 380bp, 600bp and 700bp. The PCR fragments were later cloned into pUC19 and transformed into E. coli 1M 1 09 competent cells. However, neither PCR amplification using M13 primers nor restriction digestion using BamHI obtain successful cloning results. The sequence of the PCR product was obtained through direct PCR fragment sequencing. However, the sequence derived did not show homology to other small subunit of AGPase gene when analyzed through NCBI BLAST.

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