Isolation of o-acetylserine thiol lyase genomic dna sequence from leucaena leucocephala via PCR

• Main Author: Abirame, a/p Segaran
• Format: Final Year Project Report
• Language: English; English
• Published: Universiti Malaysia Sarawak, (UNIMAS) 2015
• Subjects: SF Animal culture
• Online Access: http://ir.unimas.my/id/eprint/11412/3/Abirame%20AlP%20Segaran%2024pgs.pdf; http://ir.unimas.my/id/eprint/11412/6/Abirame%20AlP%20Segaran%20ft.pdf; http://ir.unimas.my/id/eprint/11412/

Leucaena leucocephala is widely used as a fodder for livestock animals due to its high nutritious value compared to other tropical plants. However, quality of the plant product as a livestock fodder became questionable due to the toxic effect by its degradation product, mimosine (b-N-[3-hydroxy-4-pyridone]-aaminopropionic acid) to herbivores upon ingestion. Studies suggest that inactivation or reduction of these substances will improve the quality ofL. leucocephala in animal feed production. Most of the studies have been done yet focus on reducing these substances by the use of chemicals such as FeCl3 and HCI or by the degradation enzymes extracted from ruminant microorganisms. On the other hand, the enzyme 0acetylserine thiollyase which catalyzes the synthesis ofmimosine is crucial in studies to reduce mimosine content. In this study, the leaf ofLeucaena leucocephala was used to extract the total genomic DNA. Forward and reverse primers were designed based on the O-acetylserine thiollyase mRNA sequences obtained from the NCBI database. Polymerase Chain Reaction (PCR) was performed based on the standard PCR method to amplify O-acetylserine thiollyase DNA fragment. The DNA fragment of about 1.7 kb was sent for sequencing