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KMID : 1094720130180030581
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Biotechnology and Bioprocess Engineering
2013 Volume.18 No. 3 p.581 ~ p.586
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An improved integrative transformation system for Pichia pastoris with DNA-polyethylenimine-dextran sulfate nanoparticles
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Xue Wei
Wang Hang
Pei Zuodi
Zhao Wei
Tian Yong-Sheng
Peng Ri-He
Yao Quan-Hong
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Abstract
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Pichia pastoris is a suitable host for selecting mutant enzymes, since it can secret many gene products to the medium. However, poor transformation efficiency is often encountered. This makes it difficult to obtain a large number of transformants necessary to thoroughly cover a large library. We report here that pre-coating DNA with polyethyleneimine and dextran sulfate increased gene integration significantly in Pichia electroporation. This improvement on integrative efficiency has been proved in different voltage, resistance, cell phase, and DNA concentrations. The condensed DNA nanoparticles make Pichia available as a host for screening large libraries of random mutants.
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KEYWORD
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DNA nanoparticles, integration efficiency, polyethyleneimine, yeast electroporation
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