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Title: | Electroporation of Chlorella sp. with 35 s-ap circular construct |
Authors: | Banreet Kaur Bud Singh |
Keywords: | LP 7 FST 2 2007 Banreet Kaur Bud Singh |
Issue Date: | 2007 |
Publisher: | Terengganu: Universiti Malaysia Terengganu |
Abstract: | Electroporation has been given much attention in recent years due to its easy ability to act as a tool of genetic manipulation to generate transgenic plants. The availability of essential polyunsaturated fatty acids in Ch/ore/la sp. makes it a marketable source of omega-3 and omega-6. The 35S-AP plasmid DNA was extracted from Escherichia coli. The purity of the extracted plasmid was 1.78 while the concentration was 0. 67 µg/mL. This suggests that the plasmid obtained is of high quality. The 35S-AP plasmid was amplified with PCR technique using three primer combinations. The primer combinations 35S-F/35S-R produced a band of 326 bp. The primer combinations PTE-VF1/PTE-VR2 produced a band of 617 bp. The final primer combinations 35S-F/PTE-VF1 produced a distinct band of 943 bp. A series of voltage was tested to determine the most suitable voltage by electroporation of 35S-AP circular plasmid using the Bio-Rad Micropulser electroporation apparatus . The program Ec3 with a voltage of 3.0kV was selected to electroporate Ch/ore/la sp. with 35S-AP circular plasmid. The electroporated cells were cultured on BBM with 10 µg/mL hygromycin for selection of putative recombinant Ch/ore/la sp. Sixteen putative colonies were randomly selected and transferred to BBM grid plates with 10 µg/mL and 15 µg/mL hygromycin respectively. The putative transformed colonies grew on 10 µg/mL hygromycin but no growth was observed on 15 µg/mL hygromycin. Electroporation condition needs to be optimized to su�essfully transform Ch/ore/la sp. |
URI: | http://hdl.handle.net/123456789/8393 |
Appears in Collections: | Fakulti Sains dan Teknologi |
Files in This Item:
File | Description | Size | Format | |
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LP 7 FST 2 2007 Abstract.pdf | 703.99 kB | Adobe PDF | View/Open | |
LP 7 FST 2 2007 Full Text.pdf Restricted Access | 2.79 MB | Adobe PDF | View/Open Request a copy |
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