Molecular Cloning, Isolation and Characterization of a Pectate Lyase Gene from Grape (Vitis vinifera L. cv. Bidaneh Sefid) Berry Tissue

Document Type : Research Paper

Authors

1 Associate Professor, Department of Agricultural Biotechnology, Technical and Engineering Faculty, Imam Khomeini International University, Qazvin

2 Graduate Student, Department of Agricultural Biotechnology, Technical and Engineering Faculty, Imam Khomeini International University, Qazvin

3 Assistant Professor, Department of Agricultural Biotechnology, Technical and Engineering Faculty, Imam Khomeini International University, Qazvin

Abstract

One of the important activities of the plant cell on the life cycle is to promot genes and enzyms involved in degradation of the storage components at the bigining of the fruit formation. Based on the researches, different enzymes take part in the cell wall modification and fruit tissue softening. Pectate lyase is one of the most important enzymes involved in such activities. Partial cDNA of a pectate lyase gene was isolated from grape (Vitis vinifera L. cv. Bidaneh Sefid) berry tissue by reverse transcription polymerase chain reaction (RT-PCR) technique and then was cloned into a pUC19 plasmid. Then, molecular structur, biochemical and phylogenetic characteristics were analyzed. Nucleotide sequence of the gene, called VvPel1, revealed a 429 bp long, encoding a protein of 143 amino acid residues. The calculated molecular mass and the predicted isoelectric point of the deduced polypeptide were 15.78 and 6.53 kDa, respectively. Analysis of secondary and three dimensional structures of the deduced polypeptide sequence for VvPel1 was shown that VvPel1 is similar to those of previouly reported pectate lyase proteins from other organisms. The deduced protein sequence was indicated a high similarity to pectate lyases isolated from plants such as Ricinus communis, Fragaria ananassa, Prunus persica, and Populus trichocarpa. Phylogenetic study was also demonstrated that VvPel1 gene belongs to the subgroup I of plant pectate lyases.

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Volume 3, Issue 1 - Serial Number 1
November 2012
Pages 23-34

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