Literature DB >> 23076800

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

Elke Uribe1, Meenakshi Venkatesan, David R Rose, Kathryn Vanya Ewart.   

Abstract

The 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.

Entities:  

Year:  2012        PMID: 23076800      PMCID: PMC3720964          DOI: 10.1007/s10616-012-9505-7

Source DB:  PubMed          Journal:  Cytotechnology        ISSN: 0920-9069            Impact factor:   2.058


  36 in total

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Authors:  N S Tan; M L Ng; Y H Yau; P K Chong; B Ho; J L Ding
Journal:  FASEB J       Date:  2000-09       Impact factor: 5.191

2.  Yeast-binding C-type lectin with opsonic activity from conger eel (Conger myriaster) skin mucus.

Authors:  Shigeyuki Tsutsui; Kyousuke Iwamoto; Osamu Nakamura; Tasuku Watanabe
Journal:  Mol Immunol       Date:  2006-06-05       Impact factor: 4.407

3.  Identification of a pathogen-binding lectin in salmon serum.

Authors:  K V Ewart; S C Johnson; N W Ross
Journal:  Comp Biochem Physiol C Pharmacol Toxicol Endocrinol       Date:  1999-05

4.  Enhancement of anti-Aeromonas salmonicida activity in Atlantic salmon (Salmo salar) macrophages by a mannose-binding lectin.

Authors:  C A Ottinger; S C Johnson; K V Ewart; L L Brown; N W Ross
Journal:  Comp Biochem Physiol C Pharmacol Toxicol Endocrinol       Date:  1999-05

5.  Cleavage of structural proteins during the assembly of the head of bacteriophage T4.

Authors:  U K Laemmli
Journal:  Nature       Date:  1970-08-15       Impact factor: 49.962

6.  Cystine-mediated oligomerization of the Atlantic salmon serum C-type lectin.

Authors:  David M Hudson; Neil R Mattatall; Elke Uribe; Robert C Richards; Huansheng Gong; K Vanya Ewart
Journal:  Biochim Biophys Acta       Date:  2010-11-23

7.  Cross-talk of anosmin-1, the protein implicated in X-linked Kallmann's syndrome, with heparan sulphate and urokinase-type plasminogen activator.

Authors:  Youli Hu; David González-Martínez; Soo-Hyun Kim; Pierre Marc Gilles Bouloux
Journal:  Biochem J       Date:  2004-12-15       Impact factor: 3.857

8.  Furunculosis of salmonids: vaccination attempts in rainbow trout (Salmo gairdneri) by formalin-killed germs.

Authors:  C Michel
Journal:  Ann Rech Vet       Date:  1979

9.  Insights into the mechanism of Drosophila melanogaster Golgi alpha-mannosidase II through the structural analysis of covalent reaction intermediates.

Authors:  Shin Numao; Douglas A Kuntz; Stephen G Withers; David R Rose
Journal:  J Biol Chem       Date:  2003-09-05       Impact factor: 5.157

Review 10.  The cold-shock response in cultured mammalian cells: harnessing the response for the improvement of recombinant protein production.

Authors:  Mohamed B Al-Fageeh; Rosalyn J Marchant; Martin J Carden; C Mark Smales
Journal:  Biotechnol Bioeng       Date:  2006-04-05       Impact factor: 4.530

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  3 in total

1.  Optimized expression of the antimicrobial protein Gloverin from Galleria mellonella using stably transformed Drosophila melanogaster S2 cells.

Authors:  Jan Zitzmann; Tobias Weidner; Peter Czermak
Journal:  Cytotechnology       Date:  2017-01-28       Impact factor: 2.058

2.  Srlp is crucial for the self-renewal and differentiation of germline stem cells via RpL6 signals in Drosophila testes.

Authors:  Jun Yu; Yidan Yan; Xiaojin Luan; Chen Qiao; Yuanyuan Liu; Dan Zhao; Bing Xie; Qianwen Zheng; Min Wang; Wanyin Chen; Cong Shen; Zeyu He; Xing Hu; Xiaoyan Huang; Hong Li; Qixiang Shao; Xia Chen; Bo Zheng; Jie Fang
Journal:  Cell Death Dis       Date:  2019-04-01       Impact factor: 8.469

3.  Single-cell cloning enables the selection of more productive Drosophila melanogaster S2 cells for recombinant protein expression.

Authors:  Jan Zitzmann; Christine Schreiber; Joel Eichmann; Roberto Otmar Bilz; Denise Salzig; Tobias Weidner; Peter Czermak
Journal:  Biotechnol Rep (Amst)       Date:  2018-07-03
  3 in total

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