Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus
Two clones of N1 gene derived from isolate A/Dk/Tangerang/Bbalitvet-ACIAR-TE11/2007 (H5N1) exhibit single mismatch of amino acid sequence at position 242 that is threonine and methionine for the clone #3 and #5, respectively. In order to evaluate the effect of the amino acid substitution, these clon...
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| Format: | Article |
| Language: | English |
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Bogor Agricultural University
2012-12-01
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| Series: | Hayati Journal of Biosciences |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1978301916301528 |
| _version_ | 1818109814878765056 |
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| author | RISZA HARTAWAN NI LUH PUTU INDI DHARMAYANTI KARL ROBINSON TIMOTHY MAHONY JOANNE MEERS |
| author_facet | RISZA HARTAWAN NI LUH PUTU INDI DHARMAYANTI KARL ROBINSON TIMOTHY MAHONY JOANNE MEERS |
| author_sort | RISZA HARTAWAN |
| collection | DOAJ |
| description | Two clones of N1 gene derived from isolate A/Dk/Tangerang/Bbalitvet-ACIAR-TE11/2007 (H5N1) exhibit single mismatch of amino acid sequence at position 242 that is threonine and methionine for the clone #3 and #5, respectively. In order to evaluate the effect of the amino acid substitution, these clones were inserted into two different expression vectors that are pEGFP-C1 and pcDNA-3.3 TOPO® TA cloning. Subsequently, the respective recombinant clones were transfected into eukaryotic cells, including CEF, RK13 and VERO using Lipofectamine ‘plus’ reagent. As a result, the clone #3 retaining atypical sequence showed lower expression level rather than the clone #15 in both vectors and all type of cells. The 3D conformational modelling revealed that the mutation occurs in the inner part of glycoprotein embedded within envelope or matrix. Therefore, the missense mutation seems has no effect on the antigenic properties of neuraminidase but this substitution by any means causes lethal mutagenesis in the individual gene expression by reducing level of protein transcript. |
| first_indexed | 2024-12-11T02:37:14Z |
| format | Article |
| id | doaj.art-97b0ddd297c64286a41dd76f992acf55 |
| institution | Directory Open Access Journal |
| issn | 1978-3019 |
| language | English |
| last_indexed | 2024-12-11T02:37:14Z |
| publishDate | 2012-12-01 |
| publisher | Bogor Agricultural University |
| record_format | Article |
| series | Hayati Journal of Biosciences |
| spelling | doaj.art-97b0ddd297c64286a41dd76f992acf552022-12-22T01:23:41ZengBogor Agricultural UniversityHayati Journal of Biosciences1978-30192012-12-0119416917610.4308/hjb.19.4.169Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 VirusRISZA HARTAWAN0NI LUH PUTU INDI DHARMAYANTI1KARL ROBINSON2TIMOTHY MAHONY3JOANNE MEERS4Indonesian Research Centre for Veterinary Science, Jalan R.E. Martadinata 30, Bogor 16114, IndonesiaIndonesian Research Centre for Veterinary Science, Jalan R.E. Martadinata 30, Bogor 16114, IndonesiaQueensland Alliance for Agriculture and Food Innovation, Molecular Biosciences Technologies, Research Lane, The University of Queensland, QLD 4072, AustraliaQueensland Alliance for Agriculture and Food Innovation, Molecular Biosciences Technologies, Research Lane, The University of Queensland, QLD 4072, AustraliaSchool of Veterinary Science, The University of Queensland, Gatton Campus, QLD 4343, AustraliaTwo clones of N1 gene derived from isolate A/Dk/Tangerang/Bbalitvet-ACIAR-TE11/2007 (H5N1) exhibit single mismatch of amino acid sequence at position 242 that is threonine and methionine for the clone #3 and #5, respectively. In order to evaluate the effect of the amino acid substitution, these clones were inserted into two different expression vectors that are pEGFP-C1 and pcDNA-3.3 TOPO® TA cloning. Subsequently, the respective recombinant clones were transfected into eukaryotic cells, including CEF, RK13 and VERO using Lipofectamine ‘plus’ reagent. As a result, the clone #3 retaining atypical sequence showed lower expression level rather than the clone #15 in both vectors and all type of cells. The 3D conformational modelling revealed that the mutation occurs in the inner part of glycoprotein embedded within envelope or matrix. Therefore, the missense mutation seems has no effect on the antigenic properties of neuraminidase but this substitution by any means causes lethal mutagenesis in the individual gene expression by reducing level of protein transcript.http://www.sciencedirect.com/science/article/pii/S1978301916301528missense mutationin vitro expressionH5N1N1 gene |
| spellingShingle | RISZA HARTAWAN NI LUH PUTU INDI DHARMAYANTI KARL ROBINSON TIMOTHY MAHONY JOANNE MEERS Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus Hayati Journal of Biosciences missense mutation in vitro expression H5N1 N1 gene |
| title | Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus |
| title_full | Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus |
| title_fullStr | Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus |
| title_full_unstemmed | Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus |
| title_short | Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus |
| title_sort | expression of two n1 clones with single amino acid dissimilarity of avian influenza h5n1 virus |
| topic | missense mutation in vitro expression H5N1 N1 gene |
| url | http://www.sciencedirect.com/science/article/pii/S1978301916301528 |
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