An improved integrative transformation system for Pichia pastoris with DNA-polyethylenimine-dextran sulfate nanoparticles |
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| Authors: | Wei Xue Hang Wang Zuodi Pei Wei Zhao Yong-Sheng Tian Ri-He Peng Quan-Hong Yao |
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| Affiliation: | 1667. Shanghai Key Laboratory of Agricultural Genetics and Breeding, Agro-Biotechnology Research institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China
2667. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China
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| Abstract: | 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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