Expression of recombinant Atlantic salmon serum C-type lectin in Drosophila melanogaster Schneider 2 cells

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DOIResolve DOI: http://doi.org/10.1007/s10616-012-9505-7
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TypeArticle
Journal titleCytotechnology
ISSN0920-9069
1573-0778
Volume65
Issue4
Pages513521; # of pages: 9
SubjectAtlantic salmon; C-type lectin; Drosophila; Innate immunity; Protein expression; Protein purification; Salmo salar
AbstractThe Atlantic salmon (Salmo salar) serum lectin (SSL) is a soluble C-type lectin that binds bacteria, including salmon pathogens. This lectin is a cysteine-rich oligomeric protein. Consequently, a Drosophila melanogaster expression system was evaluated for use in expressing SSL. A cDNA encoding SSL was cloned into a vector designed to express it as a fusion protein with a hexahistidine tag, under the control of the Drosophila methallothionein promoter. The resulting construct was stably transfected into Drosophila S2 cells. After CdCl2 induction, transfected S2 cells secreted recombinant SSL into the cell culture medium. A cell line derived from stably transformed polyclonal cell populations expressing SSL was used for large-scale expression of SSL. Recombinant SSL was purified from the culture medium using a two-step purification scheme involving affinity binding to yeast cells and metal-affinity chromatography. Although yields of SSL were very low, correct folding and functionality of the recombinant SSL purified in this manner was demonstrated by its ability to bind to Aeromonas salmonicida. Therefore, Drosophila S2 cells may be an ideal system for the production of SSL if yields can be increased.
Publication date
PublisherSpringer
LanguageEnglish
AffiliationNational Research Council Canada; Aquatic and Crop Resource Development
Peer reviewedYes
NRC number54062
NPARC number21268624
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Record identifier4dda93b4-d6f8-449f-9b93-afb659a81e71
Record created2013-10-28
Record modified2016-05-09
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