Document Detail


Expression of a Biologically Active GFP-α(S1)-Casein Fusion Protein in Lactococcus lactis.
MedLine Citation:
PMID:  22437853     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
In this study, we successfully developed a recombinant strain of Lactococcus lactis NZ9000 (NZ9000) that produced green fluorescent protein fused to α(S1)-casein (GFP-α(S1)Cas). A modified lactic acid bacterial vector (pNZ8148#2) was constructed by inserting genes for GFP and α(S1)-casein, a major cow's milk allergen, and the resulting vector, pNZ8148#2-GFP-α(S1)Cas, was applied to the expression of recombinant GFP-α(S1)Cas protein (rGFP-α(S1)Cas) in NZ9000. After inducing expression with nisin, the production of rGFP-α(S1)Cas was confirmed by confocal laser microscopic analysis, and the expression conditions were optimized based on fluorescent analysis and western blotting results. Moreover, the in vitro treatment of splenocytes isolated from α-casein (≥70 % α(S)-casein)-immunized mice with rGFP-α(S1)Cas resulted in increased IL-13 mRNA expression. The observed allergic activity is indicative of the Th2-cell mediated immune response and is similar to the effects induced by exposure to α-casein. Our results suggest that the expression of rGFP-α(S1)Cas in NZ9000 may facilitate in vivo applications of this system aimed at improving the specificity of immunological responses to specific milk allergen.
Authors:
Suguru Shigemori; Shinichi Yonekura; Takashi Sato; Maya Nakanishi; Hajime Otani; Takeshi Shimosato
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-3-22
Journal Detail:
Title:  Current microbiology     Volume:  -     ISSN:  1432-0991     ISO Abbreviation:  -     Publication Date:  2012 Mar 
Date Detail:
Created Date:  2012-3-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7808448     Medline TA:  Curr Microbiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Graduate School of Agriculture, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan.
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