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1.
Org Lett ; 21(18): 7435-7439, 2019 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-31509424

RESUMO

An unprecedented copper-mediated anion transformation is reported, along with selective C═C double bond cleavage and dipolar cycloaddition reaction from simple alkenes and inexpensive copper nitrate. Various transformations demonstrate the generality of this method. Further mechanistic investigation indicates a novel ionic pathway for alkene cleavage and highlights the coeffect of iodide and boric acid as additives on the inhibition of well-documented competitive nitration byproducts.

2.
Org Biomol Chem ; 17(22): 5509-5513, 2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-31112197

RESUMO

A copper nitrate-mediated [2 + 2 + 1] cycloaddition reaction was developed for the expedient synthesis of pharmacologically interesting 3-aryl substituted isoxazolines and isoxazoles through C[double bond, length as m-dash]C bond cleavage. Copper nitrate is employed as a reaction promoter and precursor of nitrile oxides. The given approach features a new mode of cycloaddition from olefinic azlactones, copper nitrate and unsaturated compounds with wide substrate scope, good functional group tolerance and operational simplicity.

3.
Res Vet Sci ; 124: 256-262, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30999161

RESUMO

Interferon-induced proteins with tetratricopeptide repeats (IFITs) are a family of proteins strongly induced downstream of type I interferon signaling. The function of IFITs has been investigated extensively in mammals. IFIT5 is the sole protein in this family found in birds and little information is available about the function of avian IFIT5. In this study, duck IFIT5 was cloned from peripheral blood mononuclear cells. Multiple amino acid sequence alignment and phylogenetic analysis showed that duck IFIT5 is highly homologous to chicken IFIT5. Tissue specificity analysis demonstrated that duck IFIT5 was ubiquitously expressed in all examined tissues of five-day-old ducklings, with the highest expression levels in heart, followed by thymus, cerebrum, liver, and lung; kidney expressed the lowest. Quantitative real-time PCR (qRT-PCR) analysis revealed that duck IFIT5 expression rapidly increased both in vitro and in vivo after stimulation with polyinosinic:polycytidylic acid [poly (I:C)] and infection with virulent duck hepatitis A virus type 3 (DHAV-3), respectively. Altogether, these results indicate that the expression of duck IFIT5 is positively correlated with viral load and may play an important role in the immune response to DHAV-3 infection. This study lays a foundation for further research into the innate antiviral immune responses of ducklings.


Assuntos
Patos/genética , Patos/imunologia , Proteínas de Neoplasias/genética , Sequência de Aminoácidos , Animais , Animais Recém-Nascidos/genética , Animais Recém-Nascidos/imunologia , Proteínas Aviárias/química , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Sequência de Bases , Clonagem Molecular , Vírus da Hepatite do Pato/fisiologia , Hepatite Viral Animal/imunologia , Proteínas de Neoplasias/química , Proteínas de Neoplasias/metabolismo , Fases de Leitura Aberta , Filogenia , Infecções por Picornaviridae/imunologia , Infecções por Picornaviridae/veterinária , Poli I-C/farmacologia , Doenças das Aves Domésticas/imunologia , Alinhamento de Sequência
4.
Org Lett ; 21(6): 1593-1597, 2019 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-30802075

RESUMO

A palladium-catalyzed multicomponent reaction of alkynes, carboxylic acids, and isocyanides has been developed with the assistance of silver salt under mild conditions. Highly functionalized captodative olefins are synthesized efficiently by this method, which can find many applications as versatile synthons in organic synthesis.

5.
Mol Immunol ; 106: 46-52, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30576951

RESUMO

The bovine IFN-ω (BoIFN-ω) multigene family is located on chromosome 8, which has 14 potential functional genes and 10 pseudogenes. After aligning 14 BoIFN-ω subtypes and assigning the most frequently occurring amino acids in each position, one artificial consensus BoIFN-ω (CoBoIFN-ω) gene was designed, optimized and synthesized. Then, CoBoIFN-ω was expressed in Pichia pastoris, which was demonstrated to have 3.94-fold and 14.3-fold higher antiviral activity against VSV on MDBK cells than that of BoIFN-ω24 and BoIFN-ω3, respectively. Besides this, CoBoIFN-ω was confirmed to have antiviral activity against VSV on BL, BT, PK-15 cells, and against BEV, BHV-1, BPIV3 on MDBK cells. Additionally, CoBoIFN-ω could bind with bovine type I IFN receptors, and then activate the promoters of NF-κB, ISRE and BoIFN-ß, and induce the transcription of ISGs and expression of Mx1 and NF-κB p65, which suggested CoBoIFN-ω exerts antiviral activity via activation of the JAK-STAT signaling pathway. Overall, this research on CoBoIFN-ω not only extends and improves consensus IFN research, but also reveals that CoBoIFN-ω has the potential to be used in the therapy of bovine viral diseases.


Assuntos
Antivirais , Herpesvirus Bovino 1/crescimento & desenvolvimento , Interferon Tipo I , Vírus da Parainfluenza 3 Bovina/crescimento & desenvolvimento , Pichia/metabolismo , Transdução de Sinais , Vesiculovirus/crescimento & desenvolvimento , Animais , Antivirais/imunologia , Antivirais/isolamento & purificação , Antivirais/metabolismo , Antivirais/farmacologia , Bovinos , Linhagem Celular , Cricetinae , Interferon Tipo I/biossíntese , Interferon Tipo I/química , Interferon Tipo I/imunologia , Interferon Tipo I/farmacologia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia
6.
Dev Comp Immunol ; 89: 44-53, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30092316

RESUMO

IFN-kappa (IFN-κ) is a type I IFN expressed by keratinocytes, monocytes and dendritic cells with important roles during the innate immune response period. This research was conducted to elaborate the evolution and characteristics of IFN-κ in poultry. Chicken IFN-κ is located on the sex-determining Z chromosome, which is greatly different from mammals. Poultry IFN-κ cluster together in a species-specific manner through positive selection pressure and share only 19-33% homology with mammalian IFN-κ and poultry other type I IFN. Both chicken and duck IFN-κ was constitutively expressed in spleen, skin, lung, and peripheral blood mononuclear cells (PBMC), as well as being significantly induced after treatment with virus in PBMC. Biologically, poultry IFN-κ has antiviral activity against VSV in chicken embryonic fibroblasts and duck embryonic fibroblasts (CEF and DEF) cells, and induces the expression of IFN stimulated genes (ISGs). After treatment with JAK1 inhibitor, the ISGs expression can be down-regulated. Overall, our research on poultry IFN-κ not only enriches the knowledge about IFN-κ but also facilitates further research on the role of type I IFNs in antiviral defense responses in poultry.

7.
Vet Res ; 49(1): 52, 2018 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-29925406

RESUMO

Duck hepatitis A virus 3 (DHAV-3), the only member of the novel genus Avihepatovirus, in the family Picornaviridae, can cause significant economic losses for duck farms in China. Reports on the pathogenicity and the antiviral molecular mechanisms of the lethal DHAV-3 strain in ducklings are inadequate and remain poorly understood. We conducted global gene expression profiling and screened differentially expressed genes (DEG) of duckling liver tissues infected with lethal DHAV-3. There were 1643 DEG and 8979 DEG when compared with mock ducklings at 12 hours post-infection (hpi) and at 48 hpi, respectively. Gene pathway analysis of DEG highlighted mainly biological processes involved in metabolic pathways, host immune responses, and viral invasion. The results may provide valuable information for us to explore the pathogenicity of the virulent DHAV-3 strain and to improve our understanding of host-virus interactions.


Assuntos
Patos , Vírus da Hepatite do Pato/fisiologia , Hepatite Viral Animal/genética , Infecções por Picornaviridae/veterinária , Doenças das Aves Domésticas/genética , Transcriptoma , Animais , Perfilação da Expressão Gênica/veterinária , Hepatite Viral Animal/virologia , Fígado/metabolismo , Infecções por Picornaviridae/genética , Infecções por Picornaviridae/virologia , Doenças das Aves Domésticas/virologia , Análise de Sequência de RNA/veterinária , Fatores de Tempo
8.
Org Biomol Chem ; 16(15): 2602-2618, 2018 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-29565088

RESUMO

Copper has been explored as an ideal candidate for replacing noble metals in organic synthesis, especially for practical large scale preparation. Recent decades have witnessed the renaissance and improvement of copper-catalyzed and copper-mediated organic reactions. Copper nitrate is a common inorganic copper salt which has been proved to be a ubiquitous reactant in organic synthesis due to its commercial availability, stability, inexpensiveness and environmentally benign nature. Copper nitrate could be used as a nitration reagent, oxidant, catalyst or promoter, and Lewis acid as well. Remarkably, great attention has been devoted to the efficient transformation of copper nitrate into functionalized or complicated compounds through various reaction types including cyclization, C-H activation, difunctionalization, nitration, rearrangement and asymmetric synthesis with chiral ligands. Further modification of copper nitrate, such as solid-supported copper nitrate or copper nitrate complexes, extends its applications in organic synthesis. The present review highlights recent advances of copper nitrate in organic synthesis, along with the mechanisms.

9.
Mol Immunol ; 95: 30-38, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29407574

RESUMO

Duck virus hepatitis caused by duck hepatitis A virus (DHAV) is an acute and contagious disease. To better understand the pathogenic mechanism of DHAV-3 in ducklings, an infection experiment was performed. Our results showed that typical symptoms were observed in the infected ducklings. DHAV-3 could infect many tissues, leading to pathological lesions, especially on the livers and spleen, and the host immune responses are activated in infection. Real-time quantitative PCR demonstrated that expression of many innate immune-related genes was mostly up-regulated in the livers and spleen, and antiviral innate immune response was established, but not sufficient to restrict the virus replication of lethal dose. Many major pattern recognition receptors (PRRs) (RIG-1, MDA5, and TLR7) are involved in the host immune response to DHAV-3, and the expression of interferon (IFNα, IFNß and IFNγ) and antiviral proteins (MX, OAS and PKR) are also up-regulated in the liver and spleen. The expression of most cytokines (IL-1ß, IL-2 and IL-6) was also up-regulated to different degrees and was various; the expression of IL-2 increased most significantly in liver. Our data provide a foundation for further study of the pathogenicity of duck virus hepatitis and extend our understanding of the immune responses of ducklings to DHAV-3 infection.

10.
Dev Comp Immunol ; 78: 91-99, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28942155

RESUMO

The multigene family of rabbit IFN-α (RbIFN-α) is located on chromosome 1, which shows seven functional genes in type I IFN locus. A novel RbIFN-α that remains unlocated in the rabbit genome was amplified and designated as the first novel rabbit IFN-α (RbIFN-αNov1), which possesses the typical molecular characteristics of type I IFNs and could be induced in RK-13 cells and peripheral blood mononuclear cells. After the mature peptide of RbIFN-αNov1 was expressed, its antiviral activity, physicochemical characteristics, and cytotoxicity were determined in vitro. Results indicated that RbIFN-αNov1 exerted a high specific antiviral activity against VSV and a low cytotoxic effect on RK-13 cells. RbIFN-αNov1 showed high sensitivity to trypsin and remained relatively stable after acid, alkali, or heat treatment. RbIFN-αNov1 could induce Mx1 expression on RK-13 cells and activate the NF-κB, ISRE and BoIFN-ß promoter activities on bovine testicular cells. Overall, our research on RbIFN-αNov1 not only enriches the knowledge about rabbit IFNs but also makes RbIFN-αNov1 have the potential to be used as an effective therapeutic agent for rabbit viral diseases.


Assuntos
Células Epiteliais/fisiologia , Interferon-alfa/genética , Leucócitos Mononucleares/fisiologia , Infecções por Rhabdoviridae/imunologia , Vírus da Estomatite Vesicular Indiana/imunologia , Animais , Antivirais/metabolismo , Linhagem Celular , Sobrevivência Celular , Clonagem Molecular , Células Epiteliais/virologia , Regulação da Expressão Gênica , Interferon-alfa/metabolismo , Interferon beta/genética , Interferon beta/metabolismo , Proteínas de Resistência a Myxovirus/genética , Proteínas de Resistência a Myxovirus/metabolismo , NF-kappa B/metabolismo , Regiões Promotoras Genéticas/genética , Estabilidade Proteica , Coelhos , Tripsina/metabolismo
11.
Res Vet Sci ; 115: 125-131, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28254416

RESUMO

Bovine interferon-omega3 (BoIFN-ω3) gene was amplified from bovine liver genomic DNA, which encodes a 195-amino acid protein containing a 23-amino acid signal peptide. Analysis of the molecular characteristics revealed that BoIFN-ω3 evolving from IFN-ω, contained four cysteine residues and five alpha helices, showing that BoIFN-ω3 presented the typical molecular characteristics of type I interferon. BoIFN-ω3 exhibited antiviral and antiproliferative activities, which exerted a protective effect against VSV in several mammalian cell lines, as well as against BEV, IBRV, and BVDV in MDBK cell. Moreover, BoIFN-ω3 was shown to be highly sensitive to trypsin, but remaining stable despite changes in pH and temperature. Additionally, BoIFN-ω3 induced the transcription of Mx1, ISG15, and ISG56 genes, as well as the expression of Mx1 protein in a time-dependent manner. These findings will be useful to further study BoIFN-ω in host's defence against infectious diseases, particularly viral infections. Furthermore, results will facilitate further research on the bovine interferon family.


Assuntos
Bovinos/fisiologia , Interferon Tipo I/química , Interferon Tipo I/metabolismo , Animais , Antivirais/farmacologia , Linhagem Celular , DNA , Fígado/química , Fígado/metabolismo , Conformação Proteica , Vírus/efeitos dos fármacos
12.
Dev Comp Immunol ; 67: 213-220, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27693540

RESUMO

A bovine interferon-kappa (BoIFN-κ) gene was amplified, which encodes a protein of 215 amino acids sharing 63% identity with human IFN-κ. BoIFN-κ was demonstrated to have antiviral and antiproliferative activities. Moreover, BoIFN-κ was shown to be highly sensitive to trypsin, however, it remained stable despite changes in pH and temperature. Result showed that BoIFN-κ can bind with bovine type I IFN receptors, and the antiviral activity can be blocked by antibodies against type I IFN receptors or BoIFN-κ. Additionally, BoIFN-κ can induce the transcription of Mx1, ISG15 and ISG56 gene, as well as the expression of Mx1 protein. The NF-κB, ISRE, and BoIFN-ß promoter can all be activated by BoIFN-κ. This study revealed that BoIFN-κ exhibits the typical characteristics of type I IFNs and exerts antiviral activity via activation of the JAK-STAT signaling pathway. Overall, these findings will enrich the current knowledge about IFN-κ and facilitate further research on the role of type I IFN in antiviral defense responses in bovine.


Assuntos
Antivirais/metabolismo , Bovinos/imunologia , Interferon Tipo I/metabolismo , Proteínas de Resistência a Myxovirus/metabolismo , Viroses/imunologia , Animais , Humanos , Imunidade , Janus Quinases/metabolismo , Proteínas de Resistência a Myxovirus/genética , NF-kappa B/genética , NF-kappa B/metabolismo , Ligação Proteica , Receptores de Interferon/metabolismo , Fatores de Transcrição STAT/metabolismo , Alinhamento de Sequência , Transdução de Sinais , Ativação Transcricional , Ubiquitinas/genética , Ubiquitinas/metabolismo
13.
Org Lett ; 18(18): 4746-9, 2016 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-27602996

RESUMO

An efficient copper nitrate mediated chloronitration reaction was developed for the direct synthesis of α-chloro-ß-nitroolefins with high regio- and stereoselectivity from simple alkynes and low toxic stannous chloride. This protocol provides a direct access to polysubstituted alkenes with operational simplicity, good functional group tolerance, and a wide substrate scope. Various applications of given products allowed the straightforward assembly of molecular complexity and indicated them as promising and valuable building blocks in organic synthesis.

14.
Chem Rec ; 16(3): 1701-14, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27230734

RESUMO

This account aims to describe our recent efforts on the synthesis and utilization of N-containing heterocycles, where transition metals participate in the synthesis. A variety of nitrogen sources, including amines, amides, hydrazones, pyrimidines, isocyanides, and copper nitrate, have been disclosed for the synthesis of diverse bioactive and pharmacologically interesting N-containing heterocycles under the participation of transition metals. The well-known nitrogen sources, such as amines and amides, were used for the construction of indoles, isatins, and quinolones. Dihydrophthalazines, isoquinolines, indazoles, and pyrazoles were obtained from hydrazones, while various pyrimidine-containing heterocycles were afforded through regioselective C-H functionalizations using pyrimidine as the directing group. Recent research has focused on the chemistry of isocyanides to achieve several kinds of heterocyclic compounds with high efficiency under the catalysis of transition metals (Pd, Rh, Mn, Cu), through oxidative cyanation reactions, sequential isocyanide insertions into C-H, N-H, or O-H bonds, and tandem radical annulation. More recently, an efficient route to isoxazolines has been reported using copper nitrate as a novel nitrogen source.

15.
Sheng Wu Gong Cheng Xue Bao ; 32(11): 1531-1538, 2016 Nov 25.
Artigo em Chinês | MEDLINE | ID: mdl-29034623

RESUMO

Immunoglobulin (Ig) is considered a part of the innate immune system and cooperates with the complementary system as the first line of defense. In this study, a monoclonal antibody (MAb) direct against the light chain of goose Ig (GoIgCL) was generated, characterized and identified in various immunoassays to detect goose Ig. An immunoaffinity chromatography column prepared with this MAb was used to separate the goose Ig from sera. After being conjugated with horseradish peroxidase (HRP), this MAb was used as the secondary antibody to evaluate the goose-specific antibody. In addition, this MAb distinguished and localized the SIg+ lymphocytes from peripheral blood lymphocytes. MAb against GoIgCL may be good candidate to detect or purify goose Ig under various conditions and as a powerful tool for humoral immunity research on goose.


Assuntos
Anticorpos Monoclonais/imunologia , Gansos/imunologia , Cadeias Leves de Imunoglobulina/imunologia , Animais , Cromatografia de Afinidade
16.
Angew Chem Int Ed Engl ; 54(30): 8795-9, 2015 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-26088206

RESUMO

An efficient method for the regioselective synthesis of pharmacologically relevant polysubstituted Δ(2)-isoxazolines is based on the copper-mediated direct transformation of simple terminal alkynes and alkenes. The overall process involves the formation of four chemical bonds with inexpensive and readily available copper nitrate trihydrate as a novel precursor of nitrile oxides. The reaction can be easily handled and proceeds under mild conditions.


Assuntos
Alcenos/química , Alquinos/química , Cobre/química , Isoxazóis/síntese química , Nitratos/química , Alcenos/síntese química , Alquinos/síntese química , Catálise , Ciclização , Isoxazóis/química , Nitrilos/síntese química , Nitrilos/química , Óxidos/síntese química , Óxidos/química , Estereoisomerismo
17.
Mol Immunol ; 66(2): 357-63, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25951414

RESUMO

A novel bovine interferon-ω (BoIFN-ω) gene, which encodes a protein of 195 amino acids with a 23-amino acid signal peptide, was amplified from bovine liver genomic DNA through PCR and named BoIFN-ω24 according to its position in the bovine genome. In this study, the recombinant protein was expressed in Escherichia coli and antiviral or antiproliferation activity was determined in vitro. Results showed that BoIFN-ω24 exhibits high antiviral activity, which can be abrogated using PAb against BoIFN-ω24, and inhibits cell proliferation. BoIFN-ω24 also presents high sensitivity to trypsin and stability at pH 2.0 or 65°C, which are typical characteristics of type I IFN. This study revealed that BoIFN-ω24 is a potential novel effective therapeutic agent and provided a basis for further research on the BoIFN-ω multigene family.


Assuntos
Antivirais/imunologia , Interferon Tipo I/imunologia , Isoformas de Proteínas/imunologia , Vesiculovirus/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/farmacologia , Antivirais/química , Antivirais/metabolismo , Antivirais/farmacologia , Sequência de Bases , Bovinos , Linhagem Celular , Proliferação de Células , Cricetulus , DNA/classificação , DNA/imunologia , DNA/isolamento & purificação , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Interferon Tipo I/química , Interferon Tipo I/genética , Interferon Tipo I/farmacologia , Fígado/química , Fígado/imunologia , Modelos Moleculares , Dados de Sequência Molecular , Filogenia , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/farmacologia , Estabilidade Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/farmacologia , Suínos , Tripsina/química , Vesiculovirus/crescimento & desenvolvimento , Vesiculovirus/imunologia
18.
Gene ; 564(2): 160-7, 2015 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-25796602

RESUMO

CD3 is one of the most important cell surface markers of T lymphocytes which play an important role in signal transmission of antigen recognition. In this study, goose CD3ε gene was cloned by touchdown PCR with the template of goose thymus cDNA. The complete open reading frame of goose CD3ε encoded 178 amino acid residues with a 21 signal peptide. Sequence alignments showed that goose CD3ε had an amino acid sequence similarity to duck (80.3%) and chicken (66.3%). The extracellular domain of goose CD3ε was efficiently expressed as fusion protein in Escherichia coli, purified by a Ni-NTA agarose column, and the purified recombinant protein was used to produce anti-GoCD3εex polyclonal antibodies. The characteristics of PAb were identified by Western blot, cellular ELISA, IFA, FCM, and LSCM analysis. These results may be useful for a better understanding of goose CD3ε and have a foundation for the study of T cell mediated immune mechanism in waterfowl.


Assuntos
Proteínas Aviárias/genética , Complexo CD3/genética , Clonagem Molecular , Gansos/genética , Sequência de Aminoácidos , Animais , Anticorpos/imunologia , Proteínas Aviárias/química , Proteínas Aviárias/metabolismo , Sequência de Bases , Complexo CD3/química , Complexo CD3/metabolismo , Citometria de Fluxo , Microscopia Confocal , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Linfócitos T/imunologia
19.
Org Biomol Chem ; 13(16): 4642-6, 2015 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-25785917

RESUMO

A novel direct transformation of hydrazones to acyloxylated pyrazoles by copper-catalyzed regioselective olefinic C(sp(2))-H bond cycloamination and acyloxylation was performed under mild conditions, which combines the formation of the pyrazole skeleton and installation of an acyloxyl group in a single step, using facile carboxylic acids as the acyloxylation reagents.


Assuntos
Cobre/química , Hidrazonas/química , Pirazóis/química , Ácidos Carboxílicos/química , Catálise , Ciclização , Desenho de Drogas , Hidrazinas/química , Ligações de Hidrogênio , Ligantes , Metais/química , Modelos Químicos , Estrutura Molecular , Compostos Orgânicos/química
20.
J Interferon Cytokine Res ; 35(3): 168-75, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25343404

RESUMO

A bovine interferon α (BoIFNα) gene that included signal sequence was amplified from bovine liver genomic DNA. The gene was named BoIFN-α1 according to the position at which the encoded gene of the bovine IFN was located in the bovine genome. The sequence included a 23-amino-acid signal peptide and a 166-amino-acid mature peptide. The structural characteristics and phylogenetic relationships of the BoIFN-α1 gene were analyzed. A recombinant mature BoIFN-α1 (rBoIFN-α1) was expressed in the yeast Pichia pastoris. Physicochemical characteristics and antiviral activity were determined in vitro. Recombinant BoIFN-α1 was found to be highly sensitive to trypsin and stable at pH 2.0 or 65°C. It also exhibited antiviral activity, which was neutralized by a rabbit anti-rBoIFNα polyclonal antibody. This study revealed that rBoIFN-α1 has the typical characteristics of IFNα and can be used for both research and industrial application.


Assuntos
Antivirais/metabolismo , Células Epiteliais/efeitos dos fármacos , Interferon-alfa/metabolismo , Fígado/fisiologia , Animais , Anticorpos Bloqueadores/metabolismo , Antivirais/química , Bovinos , Linhagem Celular , Células Epiteliais/fisiologia , Expressão Gênica , Genoma , Humanos , Interferon-alfa/química , Interferon-alfa/genética , Filogenia , Pichia/genética , Coelhos
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