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Isolation and functional characterization of buffalo (Bubalus bubalis) β-casein promoter for driving mammary epithelial cell-specific gene expression.
J Biotechnol. 2015 Mar 20; 198:53-9.JB

Abstract

Therapeutic proteins are produced in microbes, mammalian cell lines, and body fluids by applying recombinant DNA technology. They are required for compensating the deficiency of essential proteins in patients. Animal bioreactors producing such valuable bio-pharmaceuticals in body fluids have lately emerged as efficient and cost-effective expression systems. Promoters, along with other regulatory elements of genes coding for milk proteins, have been cloned from few species for directing the expression of desired proteins in the milk of farm animals. However, buffaloes, which are the second largest source of milk production in the world, have remained unexplored for such use. Since mammary epithelial cell-specific β-casein is the most abundantly expressed protein found in buffalo milk, we have isolated the promoter region and the transcriptional regulatory element along with exon 1, Intron 1 and partial exon 2 of the β-casein gene from the genome of the Indian river buffalo (Bubalus bubalis) and have characterized the same (GenBank accession no. KF612339). Mammary epithelial cells of buffalo and human (MCF7) expressed Enhanced green fluorescent protein (EGFP) upon transfection with the construct where egfp was cloned under the β-casein promoter. Transfected HEK-293 cells failed to express EGFP. Transgenic female mice generated using this construct expressed EGFP in the milk gland during lactation, without leaky expression in any other organs. This promoter also drove expression of recombinant human Interferonγ suggesting its use for expressing recombinant bio-pharmaceuticals in the milk of buffalo or other farm animals. Additionally, this may also allow breast gland-specific gene expression for remediation of breast gland-associated diseases.

Authors+Show Affiliations

Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India.Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India.Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India.Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India. Electronic address: subeer@nii.ac.in.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25678138

Citation

Ganguli, Nirmalya, et al. "Isolation and Functional Characterization of Buffalo (Bubalus Bubalis) Β-casein Promoter for Driving Mammary Epithelial Cell-specific Gene Expression." Journal of Biotechnology, vol. 198, 2015, pp. 53-9.
Ganguli N, Ganguli N, Usmani A, et al. Isolation and functional characterization of buffalo (Bubalus bubalis) β-casein promoter for driving mammary epithelial cell-specific gene expression. J Biotechnol. 2015;198:53-9.
Ganguli, N., Ganguli, N., Usmani, A., & Majumdar, S. S. (2015). Isolation and functional characterization of buffalo (Bubalus bubalis) β-casein promoter for driving mammary epithelial cell-specific gene expression. Journal of Biotechnology, 198, 53-9. https://doi.org/10.1016/j.jbiotec.2015.02.001
Ganguli N, et al. Isolation and Functional Characterization of Buffalo (Bubalus Bubalis) Β-casein Promoter for Driving Mammary Epithelial Cell-specific Gene Expression. J Biotechnol. 2015 Mar 20;198:53-9. PubMed PMID: 25678138.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation and functional characterization of buffalo (Bubalus bubalis) β-casein promoter for driving mammary epithelial cell-specific gene expression. AU - Ganguli,Nirmalya, AU - Ganguli,Nilanjana, AU - Usmani,Abul, AU - Majumdar,Subeer S, Y1 - 2015/02/09/ PY - 2014/10/17/received PY - 2015/01/24/revised PY - 2015/02/02/accepted PY - 2015/2/14/entrez PY - 2015/2/14/pubmed PY - 2015/11/14/medline KW - Animal biotechnology KW - Biopharmaceutics KW - Buffalo KW - Recombinant proteins KW - β-Casein promoter SP - 53 EP - 9 JF - Journal of biotechnology JO - J. Biotechnol. VL - 198 N2 - Therapeutic proteins are produced in microbes, mammalian cell lines, and body fluids by applying recombinant DNA technology. They are required for compensating the deficiency of essential proteins in patients. Animal bioreactors producing such valuable bio-pharmaceuticals in body fluids have lately emerged as efficient and cost-effective expression systems. Promoters, along with other regulatory elements of genes coding for milk proteins, have been cloned from few species for directing the expression of desired proteins in the milk of farm animals. However, buffaloes, which are the second largest source of milk production in the world, have remained unexplored for such use. Since mammary epithelial cell-specific β-casein is the most abundantly expressed protein found in buffalo milk, we have isolated the promoter region and the transcriptional regulatory element along with exon 1, Intron 1 and partial exon 2 of the β-casein gene from the genome of the Indian river buffalo (Bubalus bubalis) and have characterized the same (GenBank accession no. KF612339). Mammary epithelial cells of buffalo and human (MCF7) expressed Enhanced green fluorescent protein (EGFP) upon transfection with the construct where egfp was cloned under the β-casein promoter. Transfected HEK-293 cells failed to express EGFP. Transgenic female mice generated using this construct expressed EGFP in the milk gland during lactation, without leaky expression in any other organs. This promoter also drove expression of recombinant human Interferonγ suggesting its use for expressing recombinant bio-pharmaceuticals in the milk of buffalo or other farm animals. Additionally, this may also allow breast gland-specific gene expression for remediation of breast gland-associated diseases. SN - 1873-4863 UR - https://www.unboundmedicine.com/medline/citation/25678138/Isolation_and_functional_characterization_of_buffalo__Bubalus_bubalis__β_casein_promoter_for_driving_mammary_epithelial_cell_specific_gene_expression_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0168-1656(15)00042-5 DB - PRIME DP - Unbound Medicine ER -