Jurusan Ilmu Kelautan, Fakultas Perikanan dan Ilmu Kelautan, Universitas Diponegoro, Indonesia
BibTex Citation Data :
@article{IK.IJMS19361, author = {Agus Sabdono}, title = {Deteksi Transfer Plasmid Pendegradasi 2,4-D (pPP202) pada Bakteri Escherichia coli dH5α dengan Menggunakan Indikator Media EMBA}, journal = {ILMU KELAUTAN: Indonesian Journal of Marine Sciences}, volume = {7}, number = {3}, year = {2009}, keywords = {}, abstract = { Plasmid (pPP202) dari Vibrio natriegens diintroduksi secara laboratoris dengan elektrotransformasi pada inang bakteri Escherichia coli dH5 α . Pulsed Field Gel Electrophoresis (PFGE) digunakan untuk mengisolasi plasmid DNA. Plasmid pPP202 mengandung gen yang menyandi degradasi parsial senyawa 2,4-diklorofenosi asetat (2,4-D). Bakteri E. coli dH5α tidak memiliki gen yang diperlukan untuk meneralisasi 2,4-D pada khromosomnya. Asumsi tersebut memungkinkan untuk dilakukannya studi transfergen dengan menyeleksi transconjugant pada media indikator EMBA yang mengandung 2,4-D sebagai sumber karbon. Sehingga isolat yang mengandung plasmid pPP202 pada inang E. coli bisa dideteksi dengan melihat kesamaan warna dengan koloni V . natriegens PP202 yang berwarna merah. Selanjutnya , transconjugant tersebut diuji lagi pada media Zobel 12216E + 200 ppm 2,4-D yang dibandingkan dengan koloni negatif yang berwarna putih. Kata kunci : plasmid; transfergen; transconjugant A plasmid (pPP202) of Vibrio natriegens was introduced on host Escherichia coli dH5α, with electrotransformation Pulsed Field Gel Electrophoresis (PFGE) was used to isolate the DNA plasmid. Genes on this plasmid encode partiaI 2.4-dichlorophenpxyacetic acid (2.4-D) degradation. The E. coli dH5α lacks the chromosomal genes necessary for mineralization of 2.4-D, and this fact allows presumptive transconjugants obtained in gene transfer studies to be selected by plating on EMBA indicator media containing 2.4-D as the carbon source. Use of this approach enabled detection of plasmid pPP202 transfer to E. coli where previously it had not been detected. Thus, all of the 2,4-D-degrading isolates of E. coli that contained a plasmid pPP202 whose red colony colour was similar to the colony colour of V. natriegens Strain PP202, were considered as transconjugants. In addition, transconjugants were observed at distinct times in Zobell 2216E + 200 ppm 2.4-D that supported transconjugant populations compared to controls (white colour) in which no gene transfer was detected. Keywords: plasmid; gene transfer; transconjugant }, issn = {2406-7598}, pages = {152--157} doi = {10.14710/ik.ijms.7.3.152-157}, url = {https://ejournal.undip.ac.id/index.php/ijms/article/view/19361} }
Refworks Citation Data :
Plasmid (pPP202) dari Vibrio natriegens diintroduksi secara laboratoris dengan elektrotransformasi pada inang bakteri Escherichia coli dH5α. Pulsed Field Gel Electrophoresis (PFGE) digunakan untuk mengisolasi plasmid DNA. Plasmid pPP202 mengandung gen yang menyandi degradasi parsial senyawa 2,4-diklorofenosi asetat (2,4-D). Bakteri E. coli dH5α tidak memiliki gen yang diperlukan untuk meneralisasi 2,4-D pada khromosomnya. Asumsi tersebut memungkinkan untuk dilakukannya studi transfergen dengan menyeleksi transconjugant pada media indikator EMBA yang mengandung 2,4-D sebagai sumber karbon. Sehingga isolat yang mengandung plasmid pPP202 pada inang E. coli bisa dideteksi dengan melihat kesamaan warna dengan koloni V. natriegens PP202 yang berwarna merah. Selanjutnya, transconjugant tersebut diuji lagi pada media Zobel 12216E + 200 ppm 2,4-D yang dibandingkan dengan koloni negatif yang berwarna putih.
Kata kunci : plasmid; transfergen; transconjugant
A plasmid (pPP202) of Vibrio natriegens was introduced on host Escherichia coli dH5α, with electrotransformation Pulsed Field Gel Electrophoresis (PFGE) was used to isolate the DNA plasmid. Genes on this plasmid encode partiaI 2.4-dichlorophenpxyacetic acid (2.4-D) degradation. The E. coli dH5α lacks the chromosomal genes necessary for mineralization of 2.4-D, and this fact allows presumptive transconjugants obtained in gene transfer studies to be selected by plating on EMBA indicator media containing 2.4-D as the carbon source. Use of this approach enabled detection of plasmid pPP202 transfer to E. coli where previously it had not been detected. Thus, all of the 2,4-D-degrading isolates of E. coli that contained a plasmid pPP202 whose red colony colour was similar to the colony colour of V. natriegens Strain PP202, were considered as transconjugants. In addition, transconjugants were observed at distinct times in Zobell 2216E + 200 ppm 2.4-D that supported transconjugant populations compared to controls (white colour) in which no gene transfer was detected.
Keywords: plasmid; gene transfer; transconjugant
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