Construction of a pAED4-M2 vector for expressing avian influenza A (H9N2) virus M2 gene as a universal recombinant vaccine model
We expressed the M2 gene in prokaryotic cells using the pAED4 expression vector system to produce native and purified M2 protein as a candidate for universal recombinant vaccine against influenza A subtypes. The open reading frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was a...
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| Veröffentlicht in: | Turkish journal of veterinary & animal sciences 2009-01, Vol.33 (5), p.401-406 |
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| container_title | Turkish journal of veterinary & animal sciences |
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| creator | EBRAHIMI, SEYED MAHMOUD AGHAIYPOUR, KHOSROW NILI, HASSAN ESMAILZADEH, MAJID MIRJALILI, ALI |
| description | We expressed the M2 gene in prokaryotic cells using the pAED4 expression vector system to produce native
and purified M2 protein as a candidate for universal recombinant vaccine against influenza A subtypes. The open reading
frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was amplified by 2-step RT-PCR using specific
primers and pfu DNA polymerase. pAED4 was used as expression vector, purified PCR product digested by Nde I and
EcoR I restriction enzymes was ligated to the same digested site in the vector using T4 ligase to form pAED4-M2.
The cloned M2 gene was confirmed by PCR and restriction enzymes pattern. M2 polypeptide was produced through the
expression of this recombinant expression vector (pAED4+M2) in Escherichia coli BL21 (DE3) strain. The expressed M2
polypeptide was analyzed on SDS-PAGE and confirmed by western blotting assay. The level of 100% homology between
the N-terminal domain of H5 and H9 isolates was considerable. It seems that recombinant vaccine based on Iranian
isolate A/chicken/Iran/101/1998(H9N2) M2 protein might cover all H5 and H9 circulating in Iran and neighboring
regions significantly. Further research will be needed to evaluate the immunity of the expressed M2 protein in the lab
animal model to check the native conformational structure of this expressed protein by challenging with other influenza
isolates. |
| doi_str_mv | 10.3906/vet-0804-38 |
| format | Article |
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and purified M2 protein as a candidate for universal recombinant vaccine against influenza A subtypes. The open reading
frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was amplified by 2-step RT-PCR using specific
primers and pfu DNA polymerase. pAED4 was used as expression vector, purified PCR product digested by Nde I and
EcoR I restriction enzymes was ligated to the same digested site in the vector using T4 ligase to form pAED4-M2.
The cloned M2 gene was confirmed by PCR and restriction enzymes pattern. M2 polypeptide was produced through the
expression of this recombinant expression vector (pAED4+M2) in Escherichia coli BL21 (DE3) strain. The expressed M2
polypeptide was analyzed on SDS-PAGE and confirmed by western blotting assay. The level of 100% homology between
the N-terminal domain of H5 and H9 isolates was considerable. It seems that recombinant vaccine based on Iranian
isolate A/chicken/Iran/101/1998(H9N2) M2 protein might cover all H5 and H9 circulating in Iran and neighboring
regions significantly. Further research will be needed to evaluate the immunity of the expressed M2 protein in the lab
animal model to check the native conformational structure of this expressed protein by challenging with other influenza
isolates.</description><identifier>ISSN: 1303-6181</identifier><identifier>ISSN: 1300-0128</identifier><identifier>EISSN: 1303-6181</identifier><identifier>DOI: 10.3906/vet-0804-38</identifier><language>eng</language><publisher>TÜBİTAK</publisher><subject>Animal Genetics and Breeding ; Animal Immunology ; avian influenzavirus ; aşı geliştirme ; aşı(hayvan) ; bağışık yanıt ; gen ifadesi ; gene expression ; genes ; genler ; Hayvan Genetiği ve Islahı ; Hayvan İmmünolojisi ; Hayvanların Prion, Viral, Bakteriyel ve Fungal Patojenleri ; immune response ; immunogenetics ; immunogenetik ; influenza A ; kuş gribi virüsü ; kümes hayvanları ; poultry ; Prion, Viral, Bacterial and Fungal Pathogens of Animals ; SDS-PAGE ; vaccine development ; vaccines</subject><ispartof>Turkish journal of veterinary & animal sciences, 2009-01, Vol.33 (5), p.401-406</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids></links><search><creatorcontrib>EBRAHIMI, SEYED MAHMOUD</creatorcontrib><creatorcontrib>AGHAIYPOUR, KHOSROW</creatorcontrib><creatorcontrib>NILI, HASSAN</creatorcontrib><creatorcontrib>ESMAILZADEH, MAJID</creatorcontrib><creatorcontrib>MIRJALILI, ALI</creatorcontrib><title>Construction of a pAED4-M2 vector for expressing avian influenza A (H9N2) virus M2 gene as a universal recombinant vaccine model</title><title>Turkish journal of veterinary & animal sciences</title><description>We expressed the M2 gene in prokaryotic cells using the pAED4 expression vector system to produce native
and purified M2 protein as a candidate for universal recombinant vaccine against influenza A subtypes. The open reading
frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was amplified by 2-step RT-PCR using specific
primers and pfu DNA polymerase. pAED4 was used as expression vector, purified PCR product digested by Nde I and
EcoR I restriction enzymes was ligated to the same digested site in the vector using T4 ligase to form pAED4-M2.
The cloned M2 gene was confirmed by PCR and restriction enzymes pattern. M2 polypeptide was produced through the
expression of this recombinant expression vector (pAED4+M2) in Escherichia coli BL21 (DE3) strain. The expressed M2
polypeptide was analyzed on SDS-PAGE and confirmed by western blotting assay. The level of 100% homology between
the N-terminal domain of H5 and H9 isolates was considerable. It seems that recombinant vaccine based on Iranian
isolate A/chicken/Iran/101/1998(H9N2) M2 protein might cover all H5 and H9 circulating in Iran and neighboring
regions significantly. Further research will be needed to evaluate the immunity of the expressed M2 protein in the lab
animal model to check the native conformational structure of this expressed protein by challenging with other influenza
isolates.</description><subject>Animal Genetics and Breeding</subject><subject>Animal Immunology</subject><subject>avian influenzavirus</subject><subject>aşı geliştirme</subject><subject>aşı(hayvan)</subject><subject>bağışık yanıt</subject><subject>gen ifadesi</subject><subject>gene expression</subject><subject>genes</subject><subject>genler</subject><subject>Hayvan Genetiği ve Islahı</subject><subject>Hayvan İmmünolojisi</subject><subject>Hayvanların Prion, Viral, Bakteriyel ve Fungal Patojenleri</subject><subject>immune response</subject><subject>immunogenetics</subject><subject>immunogenetik</subject><subject>influenza A</subject><subject>kuş gribi virüsü</subject><subject>kümes hayvanları</subject><subject>poultry</subject><subject>Prion, Viral, Bacterial and Fungal Pathogens of Animals</subject><subject>SDS-PAGE</subject><subject>vaccine development</subject><subject>vaccines</subject><issn>1303-6181</issn><issn>1300-0128</issn><issn>1303-6181</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><recordid>eNpNkDFPwzAUhC0EElXpxIzkEYQCtp-TJmNVCkUqsMAc2c5zZUicyk4iYOKnE1SGDqd30t274SPknLMbKFh2O2CXsJzJBPIjMuHAIMl4zo8P_CmZxfjOGONFkaZpMSE_y9bHLvSmc62nraWK7harO5k8CTqg6dpA7Sj83AWM0fktVYNTnjpv6x79t6ILerkunsUVHVzoIx3_tuiRqjhO9d4NGKKqaUDTNtp55Ts6KGPcWGnaCuszcmJVHXH2f6fk7X71ulwnm5eHx-VikxiRsi4RnGtAOQfQRnCZClXZHCWg0SyvxLzKIUVe8KxK56B1BVJIzU2FzCouCwFTcrHf7Wv1oV1T7oJrVPgqOZMZ8DG_3ucmtDEGtAeF8o9vOfIt__iWkMMvNBxs_Q</recordid><startdate>20090101</startdate><enddate>20090101</enddate><creator>EBRAHIMI, SEYED MAHMOUD</creator><creator>AGHAIYPOUR, KHOSROW</creator><creator>NILI, HASSAN</creator><creator>ESMAILZADEH, MAJID</creator><creator>MIRJALILI, ALI</creator><general>TÜBİTAK</general><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>20090101</creationdate><title>Construction of a pAED4-M2 vector for expressing avian influenza A (H9N2) virus M2 gene as a universal recombinant vaccine model</title><author>EBRAHIMI, SEYED MAHMOUD ; AGHAIYPOUR, KHOSROW ; NILI, HASSAN ; ESMAILZADEH, MAJID ; MIRJALILI, ALI</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c250t-211b3e4733bc21452adf8e43ecb08d27d835e1916d573bbd3424b1cde0fa14923</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Animal Genetics and Breeding</topic><topic>Animal Immunology</topic><topic>avian influenzavirus</topic><topic>aşı geliştirme</topic><topic>aşı(hayvan)</topic><topic>bağışık yanıt</topic><topic>gen ifadesi</topic><topic>gene expression</topic><topic>genes</topic><topic>genler</topic><topic>Hayvan Genetiği ve Islahı</topic><topic>Hayvan İmmünolojisi</topic><topic>Hayvanların Prion, Viral, Bakteriyel ve Fungal Patojenleri</topic><topic>immune response</topic><topic>immunogenetics</topic><topic>immunogenetik</topic><topic>influenza A</topic><topic>kuş gribi virüsü</topic><topic>kümes hayvanları</topic><topic>poultry</topic><topic>Prion, Viral, Bacterial and Fungal Pathogens of Animals</topic><topic>SDS-PAGE</topic><topic>vaccine development</topic><topic>vaccines</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>EBRAHIMI, SEYED MAHMOUD</creatorcontrib><creatorcontrib>AGHAIYPOUR, KHOSROW</creatorcontrib><creatorcontrib>NILI, HASSAN</creatorcontrib><creatorcontrib>ESMAILZADEH, MAJID</creatorcontrib><creatorcontrib>MIRJALILI, ALI</creatorcontrib><collection>CrossRef</collection><jtitle>Turkish journal of veterinary & animal sciences</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>EBRAHIMI, SEYED MAHMOUD</au><au>AGHAIYPOUR, KHOSROW</au><au>NILI, HASSAN</au><au>ESMAILZADEH, MAJID</au><au>MIRJALILI, ALI</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Construction of a pAED4-M2 vector for expressing avian influenza A (H9N2) virus M2 gene as a universal recombinant vaccine model</atitle><jtitle>Turkish journal of veterinary & animal sciences</jtitle><date>2009-01-01</date><risdate>2009</risdate><volume>33</volume><issue>5</issue><spage>401</spage><epage>406</epage><pages>401-406</pages><issn>1303-6181</issn><issn>1300-0128</issn><eissn>1303-6181</eissn><abstract>We expressed the M2 gene in prokaryotic cells using the pAED4 expression vector system to produce native
and purified M2 protein as a candidate for universal recombinant vaccine against influenza A subtypes. The open reading
frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was amplified by 2-step RT-PCR using specific
primers and pfu DNA polymerase. pAED4 was used as expression vector, purified PCR product digested by Nde I and
EcoR I restriction enzymes was ligated to the same digested site in the vector using T4 ligase to form pAED4-M2.
The cloned M2 gene was confirmed by PCR and restriction enzymes pattern. M2 polypeptide was produced through the
expression of this recombinant expression vector (pAED4+M2) in Escherichia coli BL21 (DE3) strain. The expressed M2
polypeptide was analyzed on SDS-PAGE and confirmed by western blotting assay. The level of 100% homology between
the N-terminal domain of H5 and H9 isolates was considerable. It seems that recombinant vaccine based on Iranian
isolate A/chicken/Iran/101/1998(H9N2) M2 protein might cover all H5 and H9 circulating in Iran and neighboring
regions significantly. Further research will be needed to evaluate the immunity of the expressed M2 protein in the lab
animal model to check the native conformational structure of this expressed protein by challenging with other influenza
isolates.</abstract><pub>TÜBİTAK</pub><doi>10.3906/vet-0804-38</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1303-6181 |
| ispartof | Turkish journal of veterinary & animal sciences, 2009-01, Vol.33 (5), p.401-406 |
| issn | 1303-6181 1300-0128 1303-6181 |
| language | eng |
| recordid | cdi_ulakbim_primary_104631 |
| source | EZB-FREE-00999 freely available EZB journals; TÜBİTAK Scientific Journals |
| subjects | Animal Genetics and Breeding Animal Immunology avian influenzavirus aşı geliştirme aşı(hayvan) bağışık yanıt gen ifadesi gene expression genes genler Hayvan Genetiği ve Islahı Hayvan İmmünolojisi Hayvanların Prion, Viral, Bakteriyel ve Fungal Patojenleri immune response immunogenetics immunogenetik influenza A kuş gribi virüsü kümes hayvanları poultry Prion, Viral, Bacterial and Fungal Pathogens of Animals SDS-PAGE vaccine development vaccines |
| title | Construction of a pAED4-M2 vector for expressing avian influenza A (H9N2) virus M2 gene as a universal recombinant vaccine model |
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