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ZJUS-B
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)  2014, Vol. 15 Issue (2): 116-124    DOI: 10.1631/jzus.B1300213
Articles     
Improvement of a gene targeting system for genetic manipulation in Penicillium digitatum
Qian Xu, Cong-yi Zhu, Ming-shang Wang, Xue-peng Sun, Hong-ye Li
Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China
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Abstract  Penicillium digitatum is the most important pathogen of postharvest citrus. Gene targeting can be done in P. digitatum using homologous recombination via Agrobacterium tumefaciens mediated transformation (ATMT), but the frequencies are often very low. In the present study, we replaced the Ku80 homolog (a gene of the non-homologous end-joining (NHEJ) pathway) with the hygromycin resistance cassette (hph) by ATMT. No significant change in vegetative growth, conidiation, or pathogenicity was observed in Ku80-deficient strain (螖PdKu80) of P. digitatum. However, using 螖PdKu80 as a targeting strain, the gene-targeting frequencies for both genes PdbrlA and PdmpkA were significantly increased. These results suggest that Ku80 plays an important role in homologous integration and the created 螖PdKu80 strain would be a good candidate for rapid gene function analysis in P. digitatum.

Key words： Penicillium digitatum      Efficiency      Gene targeting      Non-homologous end-joining (NHEJ) pathway      Fig. S1')" href="#">Ku80

The online version of this article contains supplementary materials Fig. S1      Fig. S2')" href="#">Fig. S2      Fig. S3')" href="#">Fig. S3      Table S1

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Received: 12 August 2013      Published: 28 January 2014
CLC:  S436.661.1+1  
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Qian Xu
Xue-peng Sun
Cong-yi Zhu
Ming-shang Wang
Hong-ye Li
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Qian Xu, Cong-yi Zhu, Ming-shang Wang, Xue-peng Sun, Hong-ye Li. Improvement of a gene targeting system for genetic manipulation in Penicillium digitatum. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2014, 15(2): 116-124.

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http://www.zjujournals.com/xueshu/zjus-b/10.1631/jzus.B1300213     OR     http://www.zjujournals.com/xueshu/zjus-b/Y2014/V15/I2/116

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