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Isolation and characterization of hypoxia inducible gene connective tissue growth factor (CTGF) in Labeo rohita

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Abstract

The connective tissue growth factor gene plays important role in several biological processes and also responsive to hypoxia stress in fishes. The freshwater fish, Labeo rohita, highly cultured in Indian subcontinent for food, is reported as hypoxia sensitive but annotation and sequences of nuclear genes were not available for this species so far in the public domain, except some transcripts. In this study, an attempt was made for isolation and annotation of the CTGF gene in L. rohita using information of zebrafish from the same family. The CTGF gene sequence was obtained by aligning assembled genome of L. rohita, (NCBI BioProject ID: PRJNA437789), with the CTGF protein of zebrafish. Eight overlapping sets of forward and reverse primers from aligned region were designed for amplification of around 600 bp long successive overlapping fragments of CTGF gene in L. rohita. Assembly and annotation of overlapping fragments confirmed a complete 2421 bp long CTGF gene sequence with a full coding region that comprised of five exons between 308 and 1921 positions. This annotated CTGF gene sequence was submitted to GenBank (Acc. No. KY940466). Characterization of CTGF will be an initiative in identification of hypoxia response genes in L. rohita which may further help in understanding the mechanism of hypoxia tolerability in this species.

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Abbreviations

CTGF:

Connective tissue growth factor

HAS:

Hypoxia ancillary sequence

HIF:

Hypoxia inducible factors

HRE:

Hypoxia response element

TSS:

Transcription start site

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Acknowledgements

Authors are thankful to the Director, ICAR-NBFGR, Lucknow, for providing necessary facilities to carry out this work. Authors are also thankful to the CABin Scheme of ICAR and CABin Division of ICAR-IASRI, New Delhi, for financial assistance and guidance. First author is thankful to Dr. Satarudra Prakash Singh and Dr. Vineet Awasthi, Amity University Uttar Pradesh, Lucknow, for their continued suggestions.

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Authors and Affiliations

  1. Molecular Biology and Biotechnology Division, ICAR-National Bureau of Fish Genetic Resources, Lucknow, Uttar Pradesh, 226 002, India

    Iliyas Rashid, Vishwamitra Singh Baisvar, Mahender Singh, Ravindra Kumar, Basdeo Kushwaha & Ajey Kumar Pathak

  2. AMITY Institute of Biotechnology, AMITY University Uttar Pradesh, Lucknow, Uttar Pradesh, 226 028, India

    Iliyas Rashid & Prachi Srivastava

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  1. Iliyas Rashid
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Correspondence to Ravindra Kumar.

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Rashid, I., Baisvar, V.S., Singh, M. et al. Isolation and characterization of hypoxia inducible gene connective tissue growth factor (CTGF) in Labeo rohita. Mol Biol Rep 46, 1683–1691 (2019). https://doi.org/10.1007/s11033-019-04617-w

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  • DOI: https://doi.org/10.1007/s11033-019-04617-w

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