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Beijing Da Xue Xue Bao Yi Xue Ban. 2008 Jun 18;40(3):330-3.

[Agrobacterium tumefaciens-mediated transformation of uracil auxotroph Aspergillus fumigatus is an efficient method for target gene knockout].

[Article in Chinese]

Author information

1
Department of Dermatology, Peking University First Hospital;Research Center for Medical Mycology, Peking University, Beijing, China.

Abstract

OBJECTIVE:

To investigate the efficiency of Agrobacterium tumefaciens mediated transformation of Aspergillus fumigatus by using pyrG as a recessive selectable marker.

METHODS:

FAP1 and SHO1 genes target sequences, composed of a selectable marker pyrG and the flanking sequences of the FAP1 and the SHO1 genes, were cloned into a binary plasmid pDHt/sk, respectively. The produced plasmids were transformed into A. tumefaciens. The A. tumefaciens and uracil auxotroph A. fumigatus were cocultured in induction medium without uricil and uridine at 24 degrees C for 48 h. To inhibit growth of A. tumefaciens and to select transformants, the cultures were transferred to 37 degrees C and incubated for another 48 h.

RESULTS:

In this study, A. tumefaciens-mediated transformation of A. fumigatus produced high homologous recombination rates, which was 44% (7 of 16) for FAP1 and 35% (7 of 20) for SHO1.

CONCLUSION:

Our study showed that A. tumefaciens-medidated transformation by using pyrG as a recessive selectable marker is an efficient tool for target gene deletion of A. fumigatus.

PMID:
18560466
[Indexed for MEDLINE]
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