Cloning and characterization of a putative endopolygalacturonase cDNA from ripening mango (Mangifera indica Linn cv. Nam Dok Mai).
Suntornwat O., Lertwikoon N., Bungaruang L., Chaimanee P., Speirs J.
Author Affiliation: Department of Chemistry, Faculty of Science, Silpakorn University, Nakhon Pathom 73000, Thailand.
: 153-158
Abstract : A cDNA for mango polygalacturonase (PG) was constructed in order to study the expression of its ripening specific gene. Purified mRNA from ripening mango was used as a template for RT-PCR. Endo and exo-PG sequence from various sources obtained from the EMBL database were aligned. Conserved regions of the sequence were identified and used to design primer for PCR. PCR products were fractionated on an agarose gel and bands with appropriate sizes were eluted and ligated into a plasmid vector. One of the cDNAs isolated had a size of 811 bp and encoded a 161 amino acids open reading frames, the deduced sequence of which was related to plant PGs. A near match was the endo-PG of kiwifruit, which was 51% similar at the identity level and 67% similar when conserved substitution were allowed. This PG cDNA was used as a probe for hybridization analysis of fruit RNA preparations.