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Molecular Cloning and Characterization of Mannose Binding Lectin Homologue from Snow Trout (Schizothorax richardsonii)

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Abstract

The teleost fishes depend on their innate immune system for first line of defense. Mannose binding lectin (MBL) is a member of the collectin family of proteins that plays an important role in the innate immune response in many vertebrates including fish. The present study was thus carried out to identify and characterize MBL homologue from snow trout. The cDNA of snow trout MBL homologue (galactose binding; Sch-GBL) was cloned and characterized. The obtained cDNA (778-bp) contains 762 bp long ORF (open reading frame) and a 16 bp partial 3′UTR (untranslated region); transcripts are ubiquitously expressed in all the tissues under study. Sch-GBL encodes a 253-aa protein that clusters with that of common carp, goldfish and contain several common structural motifs. In addition, the Ka/Ks ratios of the C-type lectin domain sequences depicts that they are under negative selection. Sch-GBL transcript level increases in response to challenge to Aeromonas hydrophila infection and decreases in response to downshift in temperature. Altogether, these findings suggest that Sch-GBL play an important role in the first line of fish defense mechanisms.

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Abbreviations

Sch-GBL:

Schizothorax galctose binding lectin

MBL:

Mannose binding lectin

ORF:

Open reading frame

UTR:

Untranslated region

CRD:

Carbohydrate recognition domain

CTLs:

C-type lectins

IUCN:

International Union for Conservation of Nature

RNA:

Ribonucleic acid

cDNA:

Complementary DNA

PCR:

Polymerase chain reaction

LB:

Luria-Bertani

RACE:

Rapid amplification of cDNA ends

BLAST:

Basic local alignment search tool

qRT-PCR:

Quantitative real time PCR

PBS:

Phosphate buffered saline

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Acknowledgements

The authors are thankful to Indian Council of Agricultural Research (ICAR) for financial support under National Agricultural Innovation Project (NAIP).

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Authors and Affiliations

  1. Molecular Genetics Laboratory, Directorate of Coldwater Fisheries Research, Indian Council of Agricultural Research, Bhimtal, Nainital, 263136, Uttarakhand, India

    Chirag Goel, Ashoktaru Barat & Prabhati Kumari Sahoo

  2. Department of Biotechnology, Kumaun University, Bhimtal Campus, Bhimtal, Nainital, 263136, Uttarakhand, India

    Veena Pande

Authors
  1. Chirag Goel
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  2. Ashoktaru Barat
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  3. Veena Pande
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  4. Prabhati Kumari Sahoo
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Corresponding author

Correspondence to Ashoktaru Barat.

Electronic supplementary material

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10930_2014_9591_MOESM1_ESM.tif

Figure S1 Multiple alignment of the deduced amino acid sequence of MBL homologues of Vertebrates. Identical amino acid residues between each other are shown by dots respectively. (TIFF 37 kb)

10930_2014_9591_MOESM2_ESM.tif

Figure S2 Sch-GBL expression in different tissues (Spleen, liver, heart, brain and muscles) in S. richardsonii. The mRNA level of Sch-GBL was normalized to that of β-actin and muscle was used as a calibrator sample. Values are shown as mean ± SE. (TIFF 515 kb)

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Goel, C., Barat, A., Pande, V. et al. Molecular Cloning and Characterization of Mannose Binding Lectin Homologue from Snow Trout (Schizothorax richardsonii). Protein J 34, 1–8 (2015). https://doi.org/10.1007/s10930-014-9591-9

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