Expression and subcellular targeting of human insulin-like growth factor binding protein-3 in transgenic tobacco plants

被引:9
|
作者
Cheung, Stanley C. K. [1 ]
Sun, Samuel S. M. [2 ]
Chan, Juliana C. N. [1 ]
Tong, Peter C. Y. [1 ]
机构
[1] Chinese Univ Hong Kong, Prince Wales Hosp, Dept Med & Therapeut, Shatin, Hong Kong, Peoples R China
[2] Chinese Univ Hong Kong, Dept Biol, Shatin, Hong Kong, Peoples R China
关键词
IGFBP-3; Transgenic tobacco; Protein targeting; Phaseolin; Codon modification; Recombinant human protein; GREEN FLUORESCENT PROTEIN; RECOMBINANT PROTEINS; CODON USAGE; VACUOLE; ANTIBODIES; PHASEOLIN; BIOPHARMACEUTICALS; BIOFACTORIES; PEPTIDE; SEEDS;
D O I
10.1007/s11248-009-9286-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Human insulin-like growth factor binding protein-3 (hIGFBP-3) is a multifunctional protein which has high affinity for insulin-like growth factor-I (IGF-I). It combines with IGF-I to form a tertiary complex in circulation, thus regulating the activity of IGF-I. Furthermore, recombinant hIGFBP-3 (rhIGFBP-3) has been found to negatively regulate cell proliferation and induce apoptosis. In this study, we have established an efficient plant bioreactor platform for mass production of rhIGFBP-3. Different expression constructs, driven by the seed-specific phaseolin promoter, were designed and transformed into tobacco plant via Agrobacterium. To enhance protein expression level, the signal peptide (SP) and the C-terminal tetrapeptide AFVY of phaseolin were used to direct rhIGFBP-3 to protein storage vacuole (PSV) in tobacco seed for stable accumulation. Western blot analysis showed that rhIGFBP-3 was successfully synthesized in transgenic tobacco seeds, with the highest protein expression of 800 mu g/g dry weight. The localization of rhIGFBP-3 in PSV was also evident by confocal immunofluorescence microscopy. Our results indicated that protein sorting sequences could benefit the expression level of rhIGFBP-3 and it is feasible to use plant as "bio-factory" to produce therapeutic recombinant proteins in large quantity.
引用
收藏
页码:943 / 951
页数:9
相关论文
共 50 条
  • [1] Expression and subcellular targeting of human insulin-like growth factor binding protein-3 in transgenic tobacco plants
    Stanley C. K. Cheung
    Samuel S. M. Sun
    Juliana C. N. Chan
    Peter C. Y. Tong
    [J]. Transgenic Research, 2009, 18 : 943 - 951
  • [2] EXPRESSION OF HUMAN INSULIN-LIKE GROWTH FACTOR-BINDING PROTEIN-3 IN TRANSGENIC MICE
    MURPHY, LJ
    MOLNAR, P
    LU, X
    HUANG, H
    [J]. JOURNAL OF MOLECULAR ENDOCRINOLOGY, 1995, 15 (03) : 293 - 303
  • [3] Insulin-like growth factor binding protein-3 expression in the human corneal epithelium
    Robertson, Danielle M.
    Ho, Su-Inn
    Hansen, Baranda S.
    Petroll, W. Matthew
    Cavanagh, H. Dwight
    [J]. EXPERIMENTAL EYE RESEARCH, 2007, 85 (04) : 492 - 501
  • [4] Impaired sensitivity to insulin and insulin-like growth factor-I in insulin-like growth factor binding protein-3 transgenic mice
    Silha, JV
    Gui, Y
    Murphy, LJ
    [J]. DIABETOLOGIA, 2002, 45 : A31 - A31
  • [5] Expression of insulin-like growth factor binding protein-3 in HELLP syndrome
    Li Wei
    Zhou Liping
    Kang Suya
    [J]. BMC Pregnancy and Childbirth, 23
  • [6] Expression of insulin-like growth factor binding protein-3 in HELLP syndrome
    Wei, Li
    Liping, Zhou
    Suya, Kang
    [J]. BMC PREGNANCY AND CHILDBIRTH, 2023, 23 (01)
  • [7] Expression of insulin-like growth factor binding protein-3 in pterygium tissue
    Wong, Y. W.
    Chew, J.
    Yang, H.
    Tan, D. T. H.
    Beuerman, R.
    [J]. BRITISH JOURNAL OF OPHTHALMOLOGY, 2006, 90 (06) : 769 - 772
  • [8] Targeting Insulin-Like Growth Factor-I and Insulin-Like Growth Factor-Binding Protein-3 Signaling Pathways
    Lee, Hyun
    Kim, So Ri
    Oh, Youngman
    Cho, Seong Ho
    Schleimer, Robert P.
    Lee, Yong Chul
    [J]. AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY, 2014, 50 (04) : 667 - 677
  • [9] Insulin-like growth factor binding protein-3: Insulin-like growth factor independence comes of age
    Cohen, P
    [J]. ENDOCRINOLOGY, 2006, 147 (05) : 2109 - 2111
  • [10] Adenoviral-mediated expression of human insulin-like growth factor-binding protein-3
    Firth, SM
    Ganeshprasad, U
    Poronnik, P
    Cook, DI
    Baxter, RC
    [J]. PROTEIN EXPRESSION AND PURIFICATION, 1999, 16 (01) : 202 - 211