Expression Of Recombinant Human Dna Topoisomerase In Pichia Pastoris And Analysis Of Its Activity
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Date
2017-11
Authors
Chan, Mooi Kwai
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
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.
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Keywords
Recombinant Human Dna Topoisomerase In Pichia Pastoris , Topoisomerase In Pichia Pastoris And Analysis Of Its Activity