Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana
The floral-dip transformation method for Arabidopsis thaliana has long been established. Following transformation, an important step is involving the selection for transgenic plants through antibiotics or herbicides. Hygromycin has been widely applied for transgenic plants selection. However, the...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Penerbit Universiti Kebangsaan Malaysia
2014
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| Online Access: | http://journalarticle.ukm.my/7296/1/43_1_13.pdf |
| _version_ | 1796930512880664576 |
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| author | Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z. |
| author_facet | Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z. |
| author_sort | Ee, S.F. |
| collection | UKM |
| description | The floral-dip transformation method for Arabidopsis thaliana has long been established. Following transformation, an
important step is involving the selection for transgenic plants through antibiotics or herbicides. Hygromycin has been widely
applied for transgenic plants selection. However, the concentrations used were varied between laboratories mostly in a range
of 10 mg/L up to 200 mg/L. In the present study, the hygromycin sensitivity test was performed on wild-type seeds to obtain
the most effective hygromycin concentration for selection of the transgenic. A standard curve on average mean of hypocotyls
lengths against hygromycin concentrations was constructed. Based on hypocotyls length and leaf colour, the most effective
concentration of hygromycin was determined in the range of 20 to 30 mg/L. For screening the transformed seeds, hygromycin
concentration at 25 mg/L was used. Non-transgenic plants demonstrated a clear decrease in hypocotyls lengths and no root
elongation, as compared to transgenic plants. The identified transgenic seedlings were further verified through polymerase
chain reaction (PCR), giving fragment with an expected size of 365 bp. |
| first_indexed | 2024-03-06T04:04:09Z |
| format | Article |
| id | ukm.eprints-7296 |
| institution | Universiti Kebangsaan Malaysia |
| language | English |
| last_indexed | 2024-03-06T04:04:09Z |
| publishDate | 2014 |
| publisher | Penerbit Universiti Kebangsaan Malaysia |
| record_format | dspace |
| spelling | ukm.eprints-72962016-12-14T06:43:40Z http://journalarticle.ukm.my/7296/ Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z. The floral-dip transformation method for Arabidopsis thaliana has long been established. Following transformation, an important step is involving the selection for transgenic plants through antibiotics or herbicides. Hygromycin has been widely applied for transgenic plants selection. However, the concentrations used were varied between laboratories mostly in a range of 10 mg/L up to 200 mg/L. In the present study, the hygromycin sensitivity test was performed on wild-type seeds to obtain the most effective hygromycin concentration for selection of the transgenic. A standard curve on average mean of hypocotyls lengths against hygromycin concentrations was constructed. Based on hypocotyls length and leaf colour, the most effective concentration of hygromycin was determined in the range of 20 to 30 mg/L. For screening the transformed seeds, hygromycin concentration at 25 mg/L was used. Non-transgenic plants demonstrated a clear decrease in hypocotyls lengths and no root elongation, as compared to transgenic plants. The identified transgenic seedlings were further verified through polymerase chain reaction (PCR), giving fragment with an expected size of 365 bp. Penerbit Universiti Kebangsaan Malaysia 2014-06 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/7296/1/43_1_13.pdf Ee, S.F. and Khairunnisa, M.B. and Zeti-Azura, M.H. and Noor Azmi, S. and Zamri, Z. (2014) Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana. Malaysian Applied Biology, 43 (1). pp. 119-123. ISSN 0126-8643 http://mabjournal.com/ |
| spellingShingle | Ee, S.F. Khairunnisa, M.B. Zeti-Azura, M.H. Noor Azmi, S. Zamri, Z. Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana |
| title | Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana |
| title_full | Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana |
| title_fullStr | Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana |
| title_full_unstemmed | Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana |
| title_short | Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana |
| title_sort | effective hygromycin concentration for selection of agrobacterium mediated transgenic arabidopsis thaliana |
| url | http://journalarticle.ukm.my/7296/1/43_1_13.pdf |
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