Leishmania as an Expression System for the Human Follicle-Stimulating Hormone
Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In th...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Kakhki, Somayeh [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2019 |
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| Schlagwörter: |
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| Anmerkung: |
© Shiraz University 2019 |
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| Übergeordnetes Werk: |
Enthalten in: Iranian journal of science and technology - Cham, Switzerland : Springer International Pubishing, 2004, 43(2019), 6 vom: 28. Aug., Seite 2735-2742 |
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| Übergeordnetes Werk: |
volume:43 ; year:2019 ; number:6 ; day:28 ; month:08 ; pages:2735-2742 |
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| DOI / URN: |
10.1007/s40995-019-00760-y |
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| 520 | |a Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. | ||
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10.1007/s40995-019-00760-y doi (DE-627)SPR038042967 (SPR)s40995-019-00760-y-e DE-627 ger DE-627 rakwb eng Kakhki, Somayeh verfasserin aut Leishmania as an Expression System for the Human Follicle-Stimulating Hormone 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Shiraz University 2019 Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. Recombinant expression (dpeaa)DE-He213 FSH hormone subunits (dpeaa)DE-He213 Arabgari, Atefeh aut Amini-Bayat, Zahra (orcid)0000-0003-2667-0486 aut Kazemi, Bahram aut Enthalten in Iranian journal of science and technology Cham, Switzerland : Springer International Pubishing, 2004 43(2019), 6 vom: 28. Aug., Seite 2735-2742 (DE-627)SPR038034816 (DE-600)2843077-3 2364-1819 nnns volume:43 year:2019 number:6 day:28 month:08 pages:2735-2742 https://dx.doi.org/10.1007/s40995-019-00760-y lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER AR 43 2019 6 28 08 2735-2742 |
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10.1007/s40995-019-00760-y doi (DE-627)SPR038042967 (SPR)s40995-019-00760-y-e DE-627 ger DE-627 rakwb eng Kakhki, Somayeh verfasserin aut Leishmania as an Expression System for the Human Follicle-Stimulating Hormone 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Shiraz University 2019 Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. Recombinant expression (dpeaa)DE-He213 FSH hormone subunits (dpeaa)DE-He213 Arabgari, Atefeh aut Amini-Bayat, Zahra (orcid)0000-0003-2667-0486 aut Kazemi, Bahram aut Enthalten in Iranian journal of science and technology Cham, Switzerland : Springer International Pubishing, 2004 43(2019), 6 vom: 28. Aug., Seite 2735-2742 (DE-627)SPR038034816 (DE-600)2843077-3 2364-1819 nnns volume:43 year:2019 number:6 day:28 month:08 pages:2735-2742 https://dx.doi.org/10.1007/s40995-019-00760-y lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER AR 43 2019 6 28 08 2735-2742 |
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10.1007/s40995-019-00760-y doi (DE-627)SPR038042967 (SPR)s40995-019-00760-y-e DE-627 ger DE-627 rakwb eng Kakhki, Somayeh verfasserin aut Leishmania as an Expression System for the Human Follicle-Stimulating Hormone 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Shiraz University 2019 Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. Recombinant expression (dpeaa)DE-He213 FSH hormone subunits (dpeaa)DE-He213 Arabgari, Atefeh aut Amini-Bayat, Zahra (orcid)0000-0003-2667-0486 aut Kazemi, Bahram aut Enthalten in Iranian journal of science and technology Cham, Switzerland : Springer International Pubishing, 2004 43(2019), 6 vom: 28. Aug., Seite 2735-2742 (DE-627)SPR038034816 (DE-600)2843077-3 2364-1819 nnns volume:43 year:2019 number:6 day:28 month:08 pages:2735-2742 https://dx.doi.org/10.1007/s40995-019-00760-y lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER AR 43 2019 6 28 08 2735-2742 |
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10.1007/s40995-019-00760-y doi (DE-627)SPR038042967 (SPR)s40995-019-00760-y-e DE-627 ger DE-627 rakwb eng Kakhki, Somayeh verfasserin aut Leishmania as an Expression System for the Human Follicle-Stimulating Hormone 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Shiraz University 2019 Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. Recombinant expression (dpeaa)DE-He213 FSH hormone subunits (dpeaa)DE-He213 Arabgari, Atefeh aut Amini-Bayat, Zahra (orcid)0000-0003-2667-0486 aut Kazemi, Bahram aut Enthalten in Iranian journal of science and technology Cham, Switzerland : Springer International Pubishing, 2004 43(2019), 6 vom: 28. Aug., Seite 2735-2742 (DE-627)SPR038034816 (DE-600)2843077-3 2364-1819 nnns volume:43 year:2019 number:6 day:28 month:08 pages:2735-2742 https://dx.doi.org/10.1007/s40995-019-00760-y lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER AR 43 2019 6 28 08 2735-2742 |
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10.1007/s40995-019-00760-y doi (DE-627)SPR038042967 (SPR)s40995-019-00760-y-e DE-627 ger DE-627 rakwb eng Kakhki, Somayeh verfasserin aut Leishmania as an Expression System for the Human Follicle-Stimulating Hormone 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Shiraz University 2019 Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. Recombinant expression (dpeaa)DE-He213 FSH hormone subunits (dpeaa)DE-He213 Arabgari, Atefeh aut Amini-Bayat, Zahra (orcid)0000-0003-2667-0486 aut Kazemi, Bahram aut Enthalten in Iranian journal of science and technology Cham, Switzerland : Springer International Pubishing, 2004 43(2019), 6 vom: 28. Aug., Seite 2735-2742 (DE-627)SPR038034816 (DE-600)2843077-3 2364-1819 nnns volume:43 year:2019 number:6 day:28 month:08 pages:2735-2742 https://dx.doi.org/10.1007/s40995-019-00760-y lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER AR 43 2019 6 28 08 2735-2742 |
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leishmania as an expression system for the human follicle-stimulating hormone |
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Leishmania as an Expression System for the Human Follicle-Stimulating Hormone |
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Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. © Shiraz University 2019 |
| abstractGer |
Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. © Shiraz University 2019 |
| abstract_unstemmed |
Abstract Human follicle-stimulating hormone (hFSH) is a heterodimeric glycoprotein consisting of a common α subunit non-covalently attached to the hormone-specific β subunit. The therapeutic application of this hormone requires large quantities of biologically active contaminant-free hormones. In the current study, secretory expression and purification of codon-optimized recombinant α and β subunits of hFSH were reported using Iranian Lizard Leishmania (I.L.L.) expression system. Synthetic codon-optimized genes of α and β subunits of hFSH were amplified by PCR and cloned into the pLEXSY-hyg2 and pLEXSY-neo2 vectors, respectively. The resulted expression vectors were digested and electroporated into ILL. PCR analysis showed that the genes of α and β subunits of hFSH were inserted into the genome of the parasite. Subunits expression was analyzed by dot-blot and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then purified and detected by Western blot analysis. Expression analysis confirmed the presence of protein bands in sizes similar to those of the natural hormone subunits. The low electrophoretic mobility and the apparent higher molecular weight of the expressed subunits were due to successful glycosylation of subunits by ILL expression system. © Shiraz University 2019 |
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Leishmania as an Expression System for the Human Follicle-Stimulating Hormone |
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https://dx.doi.org/10.1007/s40995-019-00760-y |
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Arabgari, Atefeh Amini-Bayat, Zahra Kazemi, Bahram |
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2024-07-03T15:53:10.518Z |
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