Cloning and heterologous expression of CDef1, a ripening-induced defensin from Capsicum annuum

Elaheh Maarof, Harun Sarahani, Zeti Azura Mohamed Hussein, Ismanizan Ismail, Roslinda Sajari, Sarah Jumali, Zamri Zainal

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

CDef1, a cDNA clone that encodes a defensin gene, was isolated from a cDNA library of ripening Capsicum annuum via differential screening. Sequence analysis of the cDNA clone showed 534 bp encoding a 75 amino acid polypeptide with a predicted pI of 8.1 and a molecular mass of 8.1 kDa. The deduced peptide consists of a signal sequence of 27 amino acids, followed by a defensin domain of 47 amino acids containing 8 conserved cysteine residues with a predicted mature peptide of 5.2 kDa. The deduced peptide has high sequence similarity to the family of plant defensins. CDef1 is expressed in the mesocarp at the onset of fruit ripening and continued thereafter. Structural analysis on the sequence revealed a Cys stabilized αβ motif (CSαβ) consisting of triple-stranded anti-parallel β-sheets and an α-helix organized in βαββ architecture. The predicted three-dimensional structure of CDef1 reveals two potential receptor-binding sites; one is located on the loop that connects β1 and an α element (Phe37, Lys38, Leu40 and Leu42) and another is located on the interconnecting loop of β2 and β3 (Ile62, Phe64 and Leu66). CDef1 was expressed using the prokaryotic Escherichia coli expression system with a 47 kDa fusion peptide.

Original languageEnglish
Pages (from-to)271-276
Number of pages6
JournalAustralian Journal of Crop Science
Volume5
Issue number3
Publication statusPublished - Mar 2011

Fingerprint

Capsicum annuum
molecular cloning
ripening
peptides
amino acids
clones
mesocarp
signal peptide
cDNA libraries
cysteine
binding sites
polypeptides
sequence analysis
molecular weight
Escherichia coli
screening
receptors
fruits
genes

Keywords

  • 3D structure
  • CDef1 gene
  • CDef1 protein
  • Defensin
  • Fruit ripening
  • Recombinant expression

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

Cite this

Cloning and heterologous expression of CDef1, a ripening-induced defensin from Capsicum annuum. / Maarof, Elaheh; Sarahani, Harun; Mohamed Hussein, Zeti Azura; Ismail, Ismanizan; Sajari, Roslinda; Jumali, Sarah; Zainal, Zamri.

In: Australian Journal of Crop Science, Vol. 5, No. 3, 03.2011, p. 271-276.

Research output: Contribution to journalArticle

@article{130d1afcaa4f4a44ac2a12b0b84a3052,
title = "Cloning and heterologous expression of CDef1, a ripening-induced defensin from Capsicum annuum",
abstract = "CDef1, a cDNA clone that encodes a defensin gene, was isolated from a cDNA library of ripening Capsicum annuum via differential screening. Sequence analysis of the cDNA clone showed 534 bp encoding a 75 amino acid polypeptide with a predicted pI of 8.1 and a molecular mass of 8.1 kDa. The deduced peptide consists of a signal sequence of 27 amino acids, followed by a defensin domain of 47 amino acids containing 8 conserved cysteine residues with a predicted mature peptide of 5.2 kDa. The deduced peptide has high sequence similarity to the family of plant defensins. CDef1 is expressed in the mesocarp at the onset of fruit ripening and continued thereafter. Structural analysis on the sequence revealed a Cys stabilized αβ motif (CSαβ) consisting of triple-stranded anti-parallel β-sheets and an α-helix organized in βαββ architecture. The predicted three-dimensional structure of CDef1 reveals two potential receptor-binding sites; one is located on the loop that connects β1 and an α element (Phe37, Lys38, Leu40 and Leu42) and another is located on the interconnecting loop of β2 and β3 (Ile62, Phe64 and Leu66). CDef1 was expressed using the prokaryotic Escherichia coli expression system with a 47 kDa fusion peptide.",
keywords = "3D structure, CDef1 gene, CDef1 protein, Defensin, Fruit ripening, Recombinant expression",
author = "Elaheh Maarof and Harun Sarahani and {Mohamed Hussein}, {Zeti Azura} and Ismanizan Ismail and Roslinda Sajari and Sarah Jumali and Zamri Zainal",
year = "2011",
month = "3",
language = "English",
volume = "5",
pages = "271--276",
journal = "Australian Journal of Crop Science",
issn = "1835-2693",
publisher = "Southern Cross Publishing and Printing Pty Ltd",
number = "3",

}

TY - JOUR

T1 - Cloning and heterologous expression of CDef1, a ripening-induced defensin from Capsicum annuum

AU - Maarof, Elaheh

AU - Sarahani, Harun

AU - Mohamed Hussein, Zeti Azura

AU - Ismail, Ismanizan

AU - Sajari, Roslinda

AU - Jumali, Sarah

AU - Zainal, Zamri

PY - 2011/3

Y1 - 2011/3

N2 - CDef1, a cDNA clone that encodes a defensin gene, was isolated from a cDNA library of ripening Capsicum annuum via differential screening. Sequence analysis of the cDNA clone showed 534 bp encoding a 75 amino acid polypeptide with a predicted pI of 8.1 and a molecular mass of 8.1 kDa. The deduced peptide consists of a signal sequence of 27 amino acids, followed by a defensin domain of 47 amino acids containing 8 conserved cysteine residues with a predicted mature peptide of 5.2 kDa. The deduced peptide has high sequence similarity to the family of plant defensins. CDef1 is expressed in the mesocarp at the onset of fruit ripening and continued thereafter. Structural analysis on the sequence revealed a Cys stabilized αβ motif (CSαβ) consisting of triple-stranded anti-parallel β-sheets and an α-helix organized in βαββ architecture. The predicted three-dimensional structure of CDef1 reveals two potential receptor-binding sites; one is located on the loop that connects β1 and an α element (Phe37, Lys38, Leu40 and Leu42) and another is located on the interconnecting loop of β2 and β3 (Ile62, Phe64 and Leu66). CDef1 was expressed using the prokaryotic Escherichia coli expression system with a 47 kDa fusion peptide.

AB - CDef1, a cDNA clone that encodes a defensin gene, was isolated from a cDNA library of ripening Capsicum annuum via differential screening. Sequence analysis of the cDNA clone showed 534 bp encoding a 75 amino acid polypeptide with a predicted pI of 8.1 and a molecular mass of 8.1 kDa. The deduced peptide consists of a signal sequence of 27 amino acids, followed by a defensin domain of 47 amino acids containing 8 conserved cysteine residues with a predicted mature peptide of 5.2 kDa. The deduced peptide has high sequence similarity to the family of plant defensins. CDef1 is expressed in the mesocarp at the onset of fruit ripening and continued thereafter. Structural analysis on the sequence revealed a Cys stabilized αβ motif (CSαβ) consisting of triple-stranded anti-parallel β-sheets and an α-helix organized in βαββ architecture. The predicted three-dimensional structure of CDef1 reveals two potential receptor-binding sites; one is located on the loop that connects β1 and an α element (Phe37, Lys38, Leu40 and Leu42) and another is located on the interconnecting loop of β2 and β3 (Ile62, Phe64 and Leu66). CDef1 was expressed using the prokaryotic Escherichia coli expression system with a 47 kDa fusion peptide.

KW - 3D structure

KW - CDef1 gene

KW - CDef1 protein

KW - Defensin

KW - Fruit ripening

KW - Recombinant expression

UR - http://www.scopus.com/inward/record.url?scp=79955966208&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79955966208&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:79955966208

VL - 5

SP - 271

EP - 276

JO - Australian Journal of Crop Science

JF - Australian Journal of Crop Science

SN - 1835-2693

IS - 3

ER -