DC 欄位 | 值 | 語言 |
dc.contributor | 生化科技學系 | en |
dc.contributor.author | Kuo, C.-Y. | en_US |
dc.contributor.author | CHING-TSAN HUANG | en_US |
dc.creator | Kuo, C.-Y.;Huang, C.-T. | en |
dc.date | 2008-02 | - |
dc.date.accessioned | 2008-02-26T02:09:37Z | - |
dc.date.accessioned | 2018-07-06T02:23:39Z | - |
dc.date.available | 2008-02-26T02:09:37Z | - |
dc.date.available | 2018-07-06T02:23:39Z | - |
dc.date.issued | 2008-02 | - |
dc.identifier.issn | 0167-7012 | - |
dc.identifier.uri | http://ntur.lib.ntu.edu.tw//handle/246246/64142 | - |
dc.description.abstract | A simple and reliable mushroom transformation procedure based on electroporation of basidiospores or mycelial fragments was developed. This method eliminated the problem of protoplast preparation, the transformation efficiency were 30-150 transformants per mug DNA and the hygromycin resistant marker gene and gus were expressed in Lentinula edodes successfully. No false positive antibiotic-resistant cultures were detected by PCR amplification and the beta-glucuronidase (GUS) expression was maintained stable during mitotic cell division without selection pressure for more than 6 months. Southern analysis of transformants indicated the integration of gene might occur by non-homologous recombination. Using the glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter with the first intron of gpd gene, the average GUS activity in L. edodes reached 144.6+/-3.9 U mg(-1) soluble protein, while only 30.1+/-0.7 U mg(-1) soluble protein was detected for those without the intron. The percentage of GUS in total soluble protein was 5.67x10(-4) (0.06%) for the transformant with the highest GUS activity. This rapid and convenient electroporation procedure offers a new approach for the genetic manipulation and tool to tag genes of important edible mushroom species. | en |
dc.format.extent | 689293 bytes | - |
dc.format.mimetype | application/pdf | - |
dc.language | en_US | en |
dc.language.iso | en_US | - |
dc.relation | Journal of Microbiological Methods 72(2):111-115 | en |
dc.relation.ispartof | Journal of Microbiological Methods | en_US |
dc.subject | Electroporation | en |
dc.subject | β-glucuronidase | en |
dc.subject | Heterologous expression | en |
dc.subject | Lentinula edodes | en |
dc.title | A reliable transformation method and heterologous expression of β-glucuronidase in Lentinula edodes | en |
dc.identifier.doi | 10.1016/j.mimet.2007.11.006 | - |
dc.identifier.scopus | 2-s2.0-38049049926 | - |
dc.relation.pages | 111-115 | - |
dc.relation.journalvolume | 72 | - |
dc.relation.journalissue | 2 | - |
dc.identifier.uri.fulltext | http://ntur.lib.ntu.edu.tw/bitstream/246246/64142/1/08JMM72(111).pdf | - |
item.fulltext | with fulltext | - |
item.languageiso639-1 | en_US | - |
item.grantfulltext | open | - |
crisitem.author.dept | Biochemical Science and Technology | - |
crisitem.author.orcid | 0000-0003-1019-784X | - |
crisitem.author.parentorg | College of Life Science | - |
顯示於: | 生化科技學系
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