Advertisement

Cloning and Characterization of Legumin Storage Protein Gene of Chickpea (Cicer arietinum L)

  • Ajin D. Mandaokar
  • Kirpa R. Koundal
Article

Abstract

Genomic DNA clones coding for legumin storage protein were isolated from chickpea (Cicer arietinum L.) subgenomic library using pea legumin cDNAs as a probe. The complete nucleotide sequence of the legumin gene (leg3) was determined. Sequence analysis indicated that leg3 contains 2022 bp long structural gene interrupted by three small introns. The gene encodes a polypeptide of 496 amino acids including a signal peptide of 21 amino acid residues and a-b subunit cleavage site. Leg3 polypeptide is rich in amide amino acids and contains 5 methionine and 6 cysteine amino acids. DNA sequence comparison showed about 70–80% identity with legumin genes from pea, soybean and broad bean. The 5’ flanking region contains regulatory elements such as TATA and legumin box and 3’ region contains a stop codon and two polyadenylation signals. Southern hybridisation indicated the presence of multiple copies of /eg3in chickpea and estimated about 10–12 copies per haploid genome. Northern blot hybridisation showed that leg3 expresses in developing seed and not in other tissues of chickpea indicating temporal and tissue-specific expression of this gene.

Key words

chickpea seed storage protein legumin gene sequence 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Derbyshire E, Wright DJ & Boulter D, Phytochemistry, 15, (1976), 3.CrossRefGoogle Scholar
  2. 2.
    Casey R, Domoney C & Ellis N, Oxford survey of plant molecular and cell biology, 3, (1986), 1.Google Scholar
  3. 3.
    Shotwell MA & Larkins BA, In The Biochemistry of plants, Vol 15 (PK Stumpf, EE Conn, Editors), Academic Press, London (1989) pp 297–345.Google Scholar
  4. 4.
    Mittal M & Koundal KR, Ind J Agric Biochem, 4, (1990), 1.Google Scholar
  5. 5.
    Mandaokar AD, Kansal R, Koundal KR & Bansal HC, J Plant Biochem Biotech, 2, (1993), 35.Google Scholar
  6. 6.
    Dellaporta SL, Wood J & Hicks JB, Plant Mol Biol Report 1 (1983) 19.CrossRefGoogle Scholar
  7. 7.
    Koundal KR, Mehta SL & Sharma RP, Ind J exp Biol. 27, (1989), 858.Google Scholar
  8. 8.
    Stephen D, Jones C & Schofield JP, Nucl Acids Res, 18 1990) 7463.Google Scholar
  9. 9.
    Chomczynski P & Sacchl N, Anal Biochem, 162, (1989), 156.CrossRefGoogle Scholar
  10. 10.
    Mandaokar AD & Koundal KR, Ind J exp Biol, 34, (1996), 496.Google Scholar
  11. 11.
    Henikoff S, Gene. 28 (1984), 351.PubMedCrossRefGoogle Scholar
  12. 12.
    Sanger F, Nicklen S & Coulsen AN, Proc Natl Acad Sci, USA, 74, (1977), 5463.PubMedCrossRefGoogle Scholar
  13. 13.
    Sambrook J, Fritsch ZF & Maniatis T, Molecular cloning: A laboratory manual, 2nd Edn, Cold Spring Harbor Laboratory, New York (1989).Google Scholar
  14. 14.
    Lycett GW, Croy RRD, Shirsat AH & Boulter D, Nucl Acids Res, 12, (1984a), 4493.PubMedCrossRefGoogle Scholar
  15. 15.
    Lycett GW, Delauney AJ, Zhao W, Gatehouse JA, Croy RRD & Boulter D, Plant Mol Biol. 3 (1984b), 91.CrossRefGoogle Scholar
  16. 16.
    Heim U, Schubert R, Baumlein H & Wobus U, Plant Mol Biol. 13, (1989), 653.PubMedCrossRefGoogle Scholar
  17. 17.
    Nielson NC, Dickinson CD, Sho T, Than VH, Scallon BJ, Fischer RL, Sims L, Drews GN & Goldberg RB, Plant Cell. 1, (1989) 313.Google Scholar
  18. 18.
    Dickinson CD, Evan RP & Nielsen NC. Nucl Acids Res, 16, (1988) 371.PubMedCrossRefGoogle Scholar
  19. 19.
    Baumlein H, Nagy I, Villarood R, Inze D & Wobus U, The Plant J, 2, (1992), 233.Google Scholar
  20. 20.
    Joshi CP, Nucl Acids Res, 15, (1987), 9627.PubMedCrossRefGoogle Scholar
  21. 21.
    Brown JWS, Nucl Acids Res, 14, (1986), 9549.PubMedCrossRefGoogle Scholar
  22. 22.
    Gatehouse JA, Evan IM, Croy RRD & Boulter D, Phil Trans R Soc Lond Ser, B 314, (1986), 367.CrossRefGoogle Scholar
  23. 23.
    Nair L & Koundal KR, J Plant Biochem Biotech 2 (1993), 29.Google Scholar
  24. 24.
    Kumar LD, Mandaokar AD & Koundal KR, Ind J exp Biol, 37, (1999), 1117Google Scholar
  25. 25.
    Yamauchi D & Minamikawak T, J Plant Res, 111, (1998), 1.CrossRefGoogle Scholar
  26. 26.
    Koundal KR, Plant Physiol Biochem, 10, (1983), 60.Google Scholar
  27. 27.
    Domoney C & Casey R, Nucl Acids Res, 13, (1985), 687.PubMedCrossRefGoogle Scholar
  28. 28.
    Shutov AD, Kakhovskaya IlA, Brown H & Muntz K, J Mol Evol, 41, (1995), 1057.PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2000

Authors and Affiliations

  1. 1.National Research Centre on Plant BiotechnologyIndian Agricultural Research InstituteNew DelhiIndia

Personalised recommendations