Isolation of partial length of laccase cDNA from Ganoderma boninense

Noor Azmi M. S., . and Lim F. H., . and Omar A. R., . and Idris A. S., . and Nor Fakhrana I., . and Ahmad Parveez G. K., . (2011) Isolation of partial length of laccase cDNA from Ganoderma boninense. [Proceedings Paper]

Laccase plays an important role in lignification and delignification in nature. It is mainly produced by fungi especially white rot basidiomycetes that are efficient lignin degraders. A partial-length cDNA of 1000 bp encoding a laccase gene was isolated from the white-rot fungus Ganoderma boninense by PCR screening method. Degenerate primers corresponding to the conserved regions of the basidiomycetes laccases were used to amplify the cDNA fragment. Based on this partial sequence gene-specific primers were synthesized for amplification of 5;- end and 3;-end regions by rapid amplification of cDNA ends RACE. A 340 bp PCR product was amplified and cloned for 5;-end region. Sequence analysis of amplified fragment indicated the presence of 160 bp overlapping sequence between the 5;-end and middle region sequences. The sequence analysis also revealed the presence of a potential start codon ATG. BLASTX search against NCBI databases revealed that the partial-length of laccase cDNA LAC1175 was highly similar to the laccase gene of G. lucidum strain RZ 89 Polyporus brumalis 82 Trametes versicolor 81 and Pycnoporus cinnabarinus 79. The cloning and purification of Laccase gene should be useful for studying its function thus facilitating future investigation of G. boninense as a pathogenic fungus to oil palm.

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Abstract

Laccase plays an important role in lignification and delignification in nature. It is mainly produced by fungi especially white rot basidiomycetes that are efficient lignin degraders. A partial-length cDNA of 1000 bp encoding a laccase gene was isolated from the white-rot fungus Ganoderma boninense by PCR screening method. Degenerate primers corresponding to the conserved regions of the basidiomycetes laccases were used to amplify the cDNA fragment. Based on this partial sequence gene-specific primers were synthesized for amplification of 5;- end and 3;-end regions by rapid amplification of cDNA ends RACE. A 340 bp PCR product was amplified and cloned for 5;-end region. Sequence analysis of amplified fragment indicated the presence of 160 bp overlapping sequence between the 5;-end and middle region sequences. The sequence analysis also revealed the presence of a potential start codon ATG. BLASTX search against NCBI databases revealed that the partial-length of laccase cDNA LAC1175 was highly similar to the laccase gene of G. lucidum strain RZ 89 Polyporus brumalis 82 Trametes versicolor 81 and Pycnoporus cinnabarinus 79. The cloning and purification of Laccase gene should be useful for studying its function thus facilitating future investigation of G. boninense as a pathogenic fungus to oil palm.

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Item Type:	Proceedings Paper
Additional Information:	Available at Perpustakaan Sultan Abdul Samad Universiti Putra Malaysia 43400 UPM Serdang Selangor Malaysia. SB608 O27M939 2011 Call Number.
AGROVOC Term:	Oil palm
AGROVOC Term:	Plant disease control
AGROVOC Term:	Ganoderma
AGROVOC Term:	Pathogenic fungi
AGROVOC Term:	Isolation
AGROVOC Term:	DNA cloning
AGROVOC Term:	DNA sequence
AGROVOC Term:	Laccase
Geographical Term:	MALAYSIA
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	24 Apr 2025 05:16
URI:	http://webagris.upm.edu.my/id/eprint/13681

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