Stable expression of bioactive recombinant pleurocidin in a fish cell line

被引:34
作者
Brocal, I.
Falco, A.
Mas, V.
Rocha, A.
Perez, L.
Coll, J. M.
Estepa, A. [1 ]
机构
[1] Miguel Hernandez Univ, IBMC, Elche 03202, Spain
[2] INIA SIGT Biotecnol, Madrid 28040, Spain
关键词
D O I
10.1007/s00253-006-0393-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Pleurocidin (Ple), a linear cationic peptide of 25 amino acids, is a member of a larger family of antimicrobial peptides present in flatfish. Previous studies have shown that Ple displays a strong antimicrobial activity against a broad spectrum of bacteria and appears to play a role in innate host defence. In this work, the genomic sequence encoding the Ple prepropeptide has been isolated from Limanda limanda and cloned in a vector under the control of a non-viral promoter (the carp beta-actin promoter). By using this construction, expression of bioactive Ple was demonstrated in transformed fish cell lines continuously growing for more than 2 years. Furthermore, the study of Ple processing, maturation and secretion (by using fusion with green fluorescence protein) and the high bactericidal activity of the secreted recombinant Ple (detectable in cell supernatants without any concentration) are all reported here, as no other recombinant Ple or fish antimicrobial peptide have been expressed before to that extent. Such an overexpression of recombinant Ple or any other related antimicrobial peptide might improve the chances to develop new antibiotic agents, as well as to provide essential information about the mechanism of action, range of activity and the role in the innate immune response of antibiotic peptides.
引用
收藏
页码:1217 / 1228
页数:12
相关论文
共 38 条
  • [11] Identification, structure and differential expression of novel pleurocidins clustered on the genome of the winter flounder, Pseudopleuronectes americanus (Walbaum)
    Douglas, SE
    Patrzykat, A
    Pytyck, J
    Gallant, JW
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 2003, 270 (18): : 3720 - 3730
  • [12] Cloning and developmental expression of a family of pleurocidin-like antimicrobial peptides from winter flounder, Pleuronectes americanus (Walbaum)
    Douglas, SE
    Gallant, JW
    Gong, Z
    Hew, C
    [J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY, 2001, 25 (02) : 137 - 147
  • [13] GROWTH ENHANCEMENT IN TRANSGENIC ATLANTIC SALMON BY THE USE OF AN ALL FISH CHIMERIC GROWTH-HORMONE GENE CONSTRUCT
    DU, SJ
    GONG, ZY
    FLETCHER, GL
    SHEARS, MA
    KING, MJ
    IDLER, DR
    HEW, CL
    [J]. BIO-TECHNOLOGY, 1992, 10 (02): : 176 - 181
  • [14] Enhanced bacterial disease resistance of transgenic channel catfish Ictalurus punctatus possessing cecropin genes
    Dunham, RA
    Warr, GW
    Nichols, A
    Duncan, PL
    Argue, B
    Middleton, D
    Kucuktas, H
    [J]. MARINE BIOTECHNOLOGY, 2002, 4 (03) : 338 - 344
  • [15] SOME PROPERTIES OF THE EPITHELIOMA-PAPULOSUM-CYPRINI (EPC) CELL-LINE FROM CARP CYPRINUS-CARPIO
    FIJAN, N
    SULIMANOVIC, D
    BEARZOTTI, M
    MUZINIC, D
    ZWILLENBERG, LO
    CHILMONCZYK, S
    VAUTHEROT, JF
    DEKINKELIN, P
    [J]. ANNALES DE VIROLOGIE, 1983, 134 (02): : 207 - 220
  • [16] GANZ T, 1993, BLOOD, V82, P641
  • [17] The role of antimicrobial peptides in animal defenses
    Hancock, REW
    Scott, MG
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (16) : 8856 - 8861
  • [18] Cationic peptides: a new source of antibiotics
    Hancock, REW
    Lehrer, R
    [J]. TRENDS IN BIOTECHNOLOGY, 1998, 16 (02) : 82 - 88
  • [19] Isolation and characterisation of tilapia β-actin promoter and comparison of its activity with carp β-actin promoter
    Hwang, GL
    Rahman, MA
    Abdul Razak, S
    Sohm, F
    Farahmand, H
    Smith, A
    Brooks, C
    Maclean, N
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 2003, 1625 (01): : 11 - 18
  • [20] Antimicrobial peptides protect coho salmon from Vibrio anguillarum infections
    Jia, X
    Patrzykat, A
    Devlin, RH
    Ackerman, PA
    Iwama, GK
    Hancock, REW
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2000, 66 (05) : 1928 - 1932