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Selection of Candidate Endogenous Control GAPDH for Gene Expression Studies in Aspergillus nidulans
J. W. Baviskar, S. R. Khandelwal, A. G. Aniwale, & D. Akolkar

Published in: Journal for Advance Research in Applied Sciences
Volume- 4, Issue-7, pp.103-108, Dec 2017
DPI :-> 16.10089.JARAS.2017.V4I7.103108.2227



Abstract
Aspergillus nidulans is a model organism to study genes of industrial purpose. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is highly expressed protein in eukaryotic domain. Almost 5% of fraction is consist of GAPDH out of total cellular protein, and is constitutively expressed at all the stages of cell cycle. Selection of reference gene plays an important role in quantification of cloning and gene expression. The research aimed at selection and detection of reference gene in A. nidulans. GAPDH partial coding sequence of A. nidulans mRNA, was retrieved from gene bank: FJ410422.1 and was subjected to blast analysis against Aspergillus fumigatus, Aspergillus niger, Aspergillus oryzae, Aspergillus flavus and Aspergillus fischeri GAPDH. It was found to be 85%, 87%, 84%, 84% and 83% identical respectively. Conserved region of 167bp sequence was considered for amplification of GAPDH. Isolated mRNA was converted to cDNA using optimized conditions on RT-PCR. The cDNA of GAPDH was then amplified using sequence specific primers. Distinct band was observed for GAPDH and laccase gene on gel electrophoresis with stability of expression at both non induced and induced state. We focused our Study to generate a platform for utilization of GAPDH gene for fungal genes analysis. Thus we would like to report that GAPDH would be a promising control, for expression analysis of genes.

Key-Words / Index Term
Aspergillus. nidulans, GAPDH, mRNA, RT-PCR, Reference gene.

How to cite this article
J. W. Baviskar, S. R. Khandelwal, A. G. Aniwale, & D. Akolkar , “Selection of Candidate Endogenous Control GAPDH for Gene Expression Studies in Aspergillus nidulans”, Journal for Advance Research in Applied Sciences, 4, Issue-7, pp.103-108, Dec 2017. DPI:16.10089.JARAS.V4.I7.2227