Production of biologically active recombinant salmon calcitonin in Escherichia coli and fish cell line
Salmon calcitonin is a small peptide hormone synthesised and released by a specialised gland called ultimobranchial gland in fish. This hormone has been used to treat osteoporosis for over 50 years. The aim of this study was to compare the efficacy of five repeats of salmon calcitonin (5sCT) produce...
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| Veröffentlicht in: | Archives of microbiology 2025-01, Vol.207 (2), p.44 |
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| creator | Nafeez Ahmed, A. Abdul Majeed, S. Taju, G. Suryakodi, S. Mithra, S. Abdul Wazith, M. J. Kanimozhi, K. Rajkumar, V. Badhusha, A. Sahul Hameed, A. S. |
| description | Salmon calcitonin is a small peptide hormone synthesised and released by a specialised gland called ultimobranchial gland in fish. This hormone has been used to treat osteoporosis for over 50 years. The aim of this study was to compare the efficacy of five repeats of salmon calcitonin (5sCT) produced in two different hosts (bacteria and fish cell line). The 5sCT gene was synthesised and cloned in prokaryotic (pET32b(+)) and eukaryotic (pcDNA 3.1 + and pGenlenti) vectors. The pET32 b cloned plasmid was transformed into bacterial host BL 21 (DE3) and expression of recombinant 5sCT was induced by IPTG. The 5sCT cloned pcDNA 3.1 and pGenlenti plasmids were transfected using Lipofectamine 3000 in snubnose pompano fin (SPF) cell line. The expression of recombinant 5sCT protein in both hosts was confirmed by Western blot and ELISA using a polyclonal antibody raised against r-5sCT in mice. The results of Western blot and ELISA confirmed the expression of 5sCT protein in
E. coli
and SPF cells. The purified r-5cST expressed from both hosts was evaluated in mice via intramuscular injection at various dosages, and it was found that it significantly decreased serum calcium levels in mice when compared to normal mice. |
| doi_str_mv | 10.1007/s00203-024-04216-9 |
| format | Article |
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E. coli
and SPF cells. The purified r-5cST expressed from both hosts was evaluated in mice via intramuscular injection at various dosages, and it was found that it significantly decreased serum calcium levels in mice when compared to normal mice.</description><identifier>ISSN: 0302-8933</identifier><identifier>ISSN: 1432-072X</identifier><identifier>EISSN: 1432-072X</identifier><identifier>DOI: 10.1007/s00203-024-04216-9</identifier><identifier>PMID: 39862273</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Animals ; Bacteria ; Biochemistry ; Biological activity ; Biomedical and Life Sciences ; Biotechnology ; Calcitonin ; Calcitonin - genetics ; Calcitonin - metabolism ; Calcium (blood) ; Calcium - metabolism ; Cell Biology ; Cell Line ; Cloning ; Cloning, Molecular ; E coli ; Ecology ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Expression vectors ; Fish ; Fishes ; Genetic Vectors ; Life Sciences ; Mice ; Microbial Ecology ; Microbiology ; Original Paper ; Osteoporosis ; Plasmids ; Plasmids - genetics ; Polyclonal antibodies ; Proteins ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Salmon</subject><ispartof>Archives of microbiology, 2025-01, Vol.207 (2), p.44</ispartof><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2024 Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</rights><rights>2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.</rights><rights>Copyright Springer Nature B.V. Feb 2025</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><orcidid>0000-0002-5038-9830 ; 0000-0002-1308-4341 ; 0000-0001-6057-4792 ; 0000-0001-6255-906X ; 0000-0002-2100-9210 ; 0000-0001-5805-3779 ; 0000-0003-2897-8425 ; 0000-0001-6561-6648 ; 0000-0001-6013-4105 ; 0000-0002-6574-548X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00203-024-04216-9$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00203-024-04216-9$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/39862273$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nafeez Ahmed, A.</creatorcontrib><creatorcontrib>Abdul Majeed, S.</creatorcontrib><creatorcontrib>Taju, G.</creatorcontrib><creatorcontrib>Suryakodi, S.</creatorcontrib><creatorcontrib>Mithra, S.</creatorcontrib><creatorcontrib>Abdul Wazith, M. J.</creatorcontrib><creatorcontrib>Kanimozhi, K.</creatorcontrib><creatorcontrib>Rajkumar, V.</creatorcontrib><creatorcontrib>Badhusha, A.</creatorcontrib><creatorcontrib>Sahul Hameed, A. S.</creatorcontrib><title>Production of biologically active recombinant salmon calcitonin in Escherichia coli and fish cell line</title><title>Archives of microbiology</title><addtitle>Arch Microbiol</addtitle><addtitle>Arch Microbiol</addtitle><description>Salmon calcitonin is a small peptide hormone synthesised and released by a specialised gland called ultimobranchial gland in fish. This hormone has been used to treat osteoporosis for over 50 years. The aim of this study was to compare the efficacy of five repeats of salmon calcitonin (5sCT) produced in two different hosts (bacteria and fish cell line). The 5sCT gene was synthesised and cloned in prokaryotic (pET32b(+)) and eukaryotic (pcDNA 3.1 + and pGenlenti) vectors. The pET32 b cloned plasmid was transformed into bacterial host BL 21 (DE3) and expression of recombinant 5sCT was induced by IPTG. The 5sCT cloned pcDNA 3.1 and pGenlenti plasmids were transfected using Lipofectamine 3000 in snubnose pompano fin (SPF) cell line. The expression of recombinant 5sCT protein in both hosts was confirmed by Western blot and ELISA using a polyclonal antibody raised against r-5sCT in mice. The results of Western blot and ELISA confirmed the expression of 5sCT protein in
E. coli
and SPF cells. The purified r-5cST expressed from both hosts was evaluated in mice via intramuscular injection at various dosages, and it was found that it significantly decreased serum calcium levels in mice when compared to normal mice.</description><subject>Animals</subject><subject>Bacteria</subject><subject>Biochemistry</subject><subject>Biological activity</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Calcitonin</subject><subject>Calcitonin - genetics</subject><subject>Calcitonin - metabolism</subject><subject>Calcium (blood)</subject><subject>Calcium - metabolism</subject><subject>Cell Biology</subject><subject>Cell Line</subject><subject>Cloning</subject><subject>Cloning, Molecular</subject><subject>E coli</subject><subject>Ecology</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Expression vectors</subject><subject>Fish</subject><subject>Fishes</subject><subject>Genetic Vectors</subject><subject>Life Sciences</subject><subject>Mice</subject><subject>Microbial Ecology</subject><subject>Microbiology</subject><subject>Original Paper</subject><subject>Osteoporosis</subject><subject>Plasmids</subject><subject>Plasmids - genetics</subject><subject>Polyclonal antibodies</subject><subject>Proteins</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Salmon</subject><issn>0302-8933</issn><issn>1432-072X</issn><issn>1432-072X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2025</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkUFLJDEQhYMoOo7-gT0sAS976bVSSXe6jyKzu4KgBwVvIZNOnEg6GTvTgv_ezIwiLBQU1Pt4POoR8oPBbwYgLzMAAq8ARQUCWVN1B2TGBMcKJD4dkhlwwKrtOD8hpzm_ADBs2_aYnPCubRAlnxF3P6Z-MhufIk2OLn0K6dkbHcI71eX8ZuloTRqWPuq4oVmHoZBFN36Too-0zCKblR29WXlNTQqe6thT5_OKGhsCDT7aM3LkdMj2_HPPyeOfxcP1v-r27u_N9dVttS7Rukr0nWgM9tAIdK1m6NAxlL2BXnYawWmwRShi7aTkWBBheydqqSXUteVz8mvvux7T62TzRg0-b1PoaNOUFWd114KQDS_oxX_oS5rGWNLtKMmwEVvq5yc1LQfbq_XoBz2-q68PFoDvgVyk-GzHbxsGatuT2vekSk9q15Pq-AcLqILT</recordid><startdate>20250125</startdate><enddate>20250125</enddate><creator>Nafeez Ahmed, A.</creator><creator>Abdul Majeed, S.</creator><creator>Taju, G.</creator><creator>Suryakodi, S.</creator><creator>Mithra, S.</creator><creator>Abdul Wazith, M. 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The 5sCT gene was synthesised and cloned in prokaryotic (pET32b(+)) and eukaryotic (pcDNA 3.1 + and pGenlenti) vectors. The pET32 b cloned plasmid was transformed into bacterial host BL 21 (DE3) and expression of recombinant 5sCT was induced by IPTG. The 5sCT cloned pcDNA 3.1 and pGenlenti plasmids were transfected using Lipofectamine 3000 in snubnose pompano fin (SPF) cell line. The expression of recombinant 5sCT protein in both hosts was confirmed by Western blot and ELISA using a polyclonal antibody raised against r-5sCT in mice. The results of Western blot and ELISA confirmed the expression of 5sCT protein in
E. coli
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| subjects | Animals Bacteria Biochemistry Biological activity Biomedical and Life Sciences Biotechnology Calcitonin Calcitonin - genetics Calcitonin - metabolism Calcium (blood) Calcium - metabolism Cell Biology Cell Line Cloning Cloning, Molecular E coli Ecology Escherichia coli - genetics Escherichia coli - metabolism Expression vectors Fish Fishes Genetic Vectors Life Sciences Mice Microbial Ecology Microbiology Original Paper Osteoporosis Plasmids Plasmids - genetics Polyclonal antibodies Proteins Recombinant Proteins - genetics Recombinant Proteins - metabolism Salmon |
| title | Production of biologically active recombinant salmon calcitonin in Escherichia coli and fish cell line |
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