The Use Of Pulse Field GelElectrophoresis For Megaplasmid
| Main Author: | Perpustakaan UGM, i-lib |
|---|---|
| Format: | Article NonPeerReviewed |
| Terbitan: |
[Yogyakarta] : Universitas Gadjah Mada
, 1994
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| Subjects: | |
| Online Access: |
https://repository.ugm.ac.id/22776/ http://i-lib.ugm.ac.id/jurnal/download.php?dataId=5698 |
Daftar Isi:
- Strain-strain bakteri Pseudomonas solanacearum E.F. Smith penyebab layu tanaman telah dilaporkan mempunyai plasmid-plasmid berukuran besar. Pada plasmid tersebut diduga terdapat gen-gen yang menentukan patogenisitas bakteri tersebut. Megaplasmid mempunyai ukuran yang sangat besar dan jumlah kopi yang rendah. Ekstraksi plasmid dengan metode lisis di luar gel menyebabkan kerusakan struktur plasmid dan tidak menghasilkan pita DNA pada gel setelah elektroforesis. Pada eksperimen ini diuji penggunaan Pulse Field Gel Electrophoresis (PFGE) untuk mengisolasi megaplasmid strain 2305L (biovar 2 berasal dart tomat di Lembang) dan strain JGP (biovar 3 berasal dart kacang tanah di Jambegede, Malang). PFGE dilarikan dengan 'ramping time' 40 � 60 detik, 'running time' 26 jam pada voltase 200 volt. Hasilnya menunjukkan bahwa kedua strain yang dicoba mempunyai megaplasmid yang dapat dipisahkan dart DNA genomnya. Ukuran megaplasmid tersebut lebih besar dart 770 kb. Abstract The strains of Pseudomonas solanacearum E.F. Smith have been reported to have .megaplasmids. The plasmid was suspected to carry genes of pathogenicity of the bacterium. The plasmids have quite big size and Iow copy number. Extraction of the plasmid out of gel causes the damage of the plasmid and no band appears on gel after the electrophoresis. This experiment was done to isolate megaplasmid of two strains of P. solanacearum strain 2305L (biovar 2 tomato isolate from Lembang) and strain JGP (biovar 3 peanut isolate from Jambegede, Malang) using Pulse Field Gel Electrophoresis (PFGE). The PFGE was run at the ramping time of 40 � 60 seconds, the running time of 26 hours and the voltage was set at 200 volt. The results suggested that the two strains of P. solanacearum have megaplasmids which could be separated from the genom. The size of the megaplasmids were bigger than 770 kb.