COMPARATIVE EXPRESSION ANALYSIS OF LACCASE GENES IN ASPERGILLUS NIDULANS MTCC-344
Main Article Content
Abstract
Laccases belong to the family of multicopper oxidoreductases. They act on large variety of substrates particularly phenols
and are biotechnologically more applicable than other enzymes. Laccases produce only carbon dioxide and water as by
product after biodegradation. Among white rot fungi Aspergillus nidulans produces three extracellular laccases. It is an ideal
organism to study genes for industrial purpose. Scanty research is available on isoenzymes of laccase belonging to A.
nidulans. In this work laccases were induced using Guaiacol as inducer. A. nidulans MTCC-344 showed highest laccase
enzyme activity of 2.09 U/ml using Guaiacol as a substrate on the 5th day after induction. This study aimed at selection
and detection of three inducible genes viz: lccB, lccC, 8lccD for expression studies. Universal primers were designed for
detection of laccases in Aspergillus family. Total mRNA was isolated and converted to cDNA using RT-PCR. Gene
expression analysis was carried out usingsemi-quantitative PCR based approach for conserved regions of lccB. lccC, lccD
sequences. Results revealed comparatively high rate of expression in lccB. Full length lccB. lccC, lccD genes were also
amplifiedand sequenced. The genes were found to be of size between 1.7 to 2 kb.BLAST analysis reviled identity to
Aspergillus family laccases. The gene sequences were submitted to Genebankwith accession number for lcc B:
MG783369,lcc C: MG783370, lcc D: MG783371. Thus the isolated genes can be a benchmark of gene expression analysis
and protein expressions studies.
and are biotechnologically more applicable than other enzymes. Laccases produce only carbon dioxide and water as by
product after biodegradation. Among white rot fungi Aspergillus nidulans produces three extracellular laccases. It is an ideal
organism to study genes for industrial purpose. Scanty research is available on isoenzymes of laccase belonging to A.
nidulans. In this work laccases were induced using Guaiacol as inducer. A. nidulans MTCC-344 showed highest laccase
enzyme activity of 2.09 U/ml using Guaiacol as a substrate on the 5th day after induction. This study aimed at selection
and detection of three inducible genes viz: lccB, lccC, 8lccD for expression studies. Universal primers were designed for
detection of laccases in Aspergillus family. Total mRNA was isolated and converted to cDNA using RT-PCR. Gene
expression analysis was carried out usingsemi-quantitative PCR based approach for conserved regions of lccB. lccC, lccD
sequences. Results revealed comparatively high rate of expression in lccB. Full length lccB. lccC, lccD genes were also
amplifiedand sequenced. The genes were found to be of size between 1.7 to 2 kb.BLAST analysis reviled identity to
Aspergillus family laccases. The gene sequences were submitted to Genebankwith accession number for lcc B:
MG783369,lcc C: MG783370, lcc D: MG783371. Thus the isolated genes can be a benchmark of gene expression analysis
and protein expressions studies.
Downloads
Download data is not yet available.
Article Details
How to Cite
Ratan, K., Waman, B., Gopalroa, A., & Bhausaheb, A. (2020). COMPARATIVE EXPRESSION ANALYSIS OF LACCASE GENES IN ASPERGILLUS NIDULANS MTCC-344. Journal of Advanced Scientific Research, 11(Suppl 6), 55-60. https://doi.org/5
Section
Research Article
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.