Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity
This study was aimed to establish a method to produce in-house recombinant human DNA topoisomerase. The DNA topoisomerase I (Topo I) and DNA topoisomerase II (Topo II) coding regions were amplified from the cDNA of breast cancer cells, MDA-MB-231. Topo I and Topo II coding region were ligated int...
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my-usm-ep.478312020-10-28T09:23:17Z Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity 2017-11 Chan, Mooi Kwai QP Physiology This study was aimed to establish a method to produce in-house recombinant human DNA topoisomerase. The DNA topoisomerase I (Topo I) and DNA topoisomerase II (Topo II) coding regions were amplified from the cDNA of breast cancer cells, MDA-MB-231. Topo I and Topo II coding region were ligated into expression vector, pPICZ--A and pPICZ--C, respectively. Subsequently, the constructed recombinant vectors were transformed into four types of Pichia pastoris strain, X-33, GS115, SMD1168H and KM71H. PCR screening for recombinant clones was performed and followed by selection of multicopy clones using agar plates containing increasing concentrations of Zeocin. The selected multicopy clones were then induced for enzymes expression in a shake flask system. Topo I was expressed relatively higher in GS115 than other Pichia strains. 2017-11 Thesis http://eprints.usm.my/47831/ http://eprints.usm.my/47831/1/EXPRESSION%20OF%20RECOMBINANT%20HUMAN%20DNA.pdf%20cut.pdf application/pdf en public phd doctoral Universiti Sains Malaysia Institut Penyelidikan Perubatan Molekul (Institute for Research in Molecular Medicine INFORM) |
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Universiti Sains Malaysia |
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USM Institutional Repository |
| language |
English |
| topic |
QP Physiology |
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QP Physiology Chan, Mooi Kwai Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity |
| description |
This study was aimed to establish a method to produce in-house recombinant human DNA topoisomerase. The DNA topoisomerase I (Topo I) and DNA topoisomerase II (Topo II) coding regions were amplified from the cDNA of breast cancer cells, MDA-MB-231. Topo I and Topo II coding region were ligated into expression vector, pPICZ--A and pPICZ--C, respectively. Subsequently, the constructed recombinant vectors were transformed into four types of Pichia pastoris strain, X-33, GS115, SMD1168H and KM71H. PCR screening for recombinant clones was performed and followed by selection of multicopy clones using agar plates containing increasing concentrations of Zeocin. The selected multicopy clones were then induced for enzymes expression in a shake flask system. Topo I was expressed relatively higher in GS115 than other Pichia strains. |
| format |
Thesis |
| qualification_name |
Doctor of Philosophy (PhD.) |
| qualification_level |
Doctorate |
| author |
Chan, Mooi Kwai |
| author_facet |
Chan, Mooi Kwai |
| author_sort |
Chan, Mooi Kwai |
| title |
Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity |
| title_short |
Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity |
| title_full |
Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity |
| title_fullStr |
Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity |
| title_full_unstemmed |
Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity |
| title_sort |
expression of recombinant human dna topoisomerase in pichia pastoris and analysis of its activity |
| granting_institution |
Universiti Sains Malaysia |
| granting_department |
Institut Penyelidikan Perubatan Molekul (Institute for Research in Molecular Medicine INFORM) |
| publishDate |
2017 |
| url |
http://eprints.usm.my/47831/1/EXPRESSION%20OF%20RECOMBINANT%20HUMAN%20DNA.pdf%20cut.pdf |
| _version_ |
1747821837787594752 |