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Efficient transformation and expression of gfp gene in the edible mushroom Pleurotus nebrodensis

Efficient transformation and expression of gfp gene in the edible mushroom Pleurotus nebrodensis

Authors :
Lin, Junfang
Zheng, Mingyao
Wang, Jie
Shu, Wei
Guo, Liqiong
Source :
Progress in Natural Science. Jul2008, Vol. 18 Issue 7, p819-824. 6p.
Publication Year :
2008

Abstract

Abstract: An efficient transformation method mediated by PEG-protoplasts was developed for the newly commercial edible mushroom Pleurotus nebrodensis. Two plasmids were used to co-transform protoplasts of P. nebrodensis. One plasmid is pAN7-1 containing a positive selectable marker gene hph conferring hygromycin B resistance. Another plasmid is pBlue-GFP containing a reporter gene gfp conferring green fluorescent protein. PCR and Southern blot analysis showed that hph gene or/and gfp gene were integrated into the genome of P. nebrodensis transformants. The transformation efficiency of the positive selectable marker gene hph was 3 transformants per microgram of plasmid pAN7-1 DNA, which was about 30 times higher than that previously reported in thoroughly studied Pleurotus species such as Pleurotus ostreatus. The transformation efficiency of the reporter gene gfp was 9 transformants per microgram of plasmid pBlu-GFP DNA. The co-transformation efficiency was 23.68%. This is the first report that a “reporter” gene, green fluorescent protein gene can be successfully stably expressed in this Pleurotus species. [Copyright &y& Elsevier]

Details

Language :
English
ISSN :
10020071
Volume :
18
Issue :
7
Database :
Academic Search Index
Journal :
Progress in Natural Science
Publication Type :
Academic Journal
Accession number :
34303746
Full Text :
https://doi.org/10.1016/j.pnsc.2008.01.015