Journal of Biosciences

, Volume 32, Issue 2, pp 241–249

cDNA cloning and expression analysis of a mannose-binding lectin from Pinellia pedatisecta

  • Juan Lin
  • Xuanwei Zhou
  • Shi Gao
  • Xiaojun Liu
  • Weisheng Wu
  • Xiaofen Sun
  • Kexuan Tang
Article
  • 110 Downloads

Abstract

Pinellia pedatisecta agglutinin (PPA) is a very basic protein that accumulates in the tuber of P. pedatisecta. PPA is a hetero-tetramer protein of 40 kDa, composed of two polypeptide chains A (about 12 kDa) and two polypeptides chains B (about 12 kDa). The full-length cDNA of PPA was cloned from P. pedatisecta using SMART RACE-PCR technology; it was 1146 bp and contained a 771 bp open reading frame (ORF) encoding a lectin precursor of 256 amino acid residues with a 24 amino acid signal peptide. The PPA precursor contained 3 mannose-binding sites (QXDXNXVXY) and two conserved domains of 43% identity, PPA-DOM1 (polypeptides A) and PPA-DOM2 (polypeptides B). PPA shared varying identities, ranging from 40% to 85%, with mannose-binding lectins from other species of plant families such as Araceae, Alliaceae, Iridaceae, Liliaceae, Amaryllidaceae and Bromeliaceae. Southern blot analysis indicated that ppa belonged to a multi-copy gene family. Expression pattern analysis revealed that ppa expressed in most tested tissues, with high expression being found in spadix, spathe and tuber. Cloning of the ppa gene not only provides a basis for further investigation of its structure, expression and regulatory mechanism, but also enables us to test its potential role in controlling pests and fungal diseases by transferring the gene into plants in the future.

Keywords

Pinellia pedatisecta Pinellia pedatisecta agglutinin (PPA) cDNA cloning mannose-binding lectin RACE expression pattern 

Abbreviations used

ORF

open reading frame

RACE

rapid amplification of cDNA ends

PPA

Pinellia pedatisecta agglutinin

PCR

polymerase chain reaction

pI

isoelectric point

RT-PCR

reverse transcriptase polymerase chain reaction

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Copyright information

© Indian Academy of Sciences 2007

Authors and Affiliations

  • Juan Lin
    • 1
  • Xuanwei Zhou
    • 2
  • Shi Gao
    • 1
  • Xiaojun Liu
    • 1
  • Weisheng Wu
    • 1
  • Xiaofen Sun
    • 1
  • Kexuan Tang
    • 1
  1. 1.State Key Laboratory of Genetic Engineering, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Life Sciences, Morgan-Tan International Center for Life SciencesFudan UniversityShanghaiPeople’s Republic of China
  2. 2.Plant Biotechnology Research Center, School of Agriculture and Biology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Life Sciences and TechnologyShanghai Jiao Tong UniversityShanghaiPeople’s Republic of China

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