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Transformation of Monascus purpureus to hygromycin B resistance with cosmid pMOcosX reduces fertility Electron. J. Biotechnol.
Lakrod,Kamolnan; Chaisrisook,Chulee; Skinner,Daniel Z.
Albino strain KB20M1 of Monascus purpureus was genetically transformed to hygromycin B resistance with cosmid pMOcosX, using biolistic bombardment. Conidia and mycelial fragments were used as the recipient material and rupture disk strengths of 6.2 and 9.3 MPa each yielded a transformed isolate from five bombardments. Southern analysis suggested a single copy of the cosmid had integrated into each of the transformants. Both of the independent transformants formed cleistothecia but ascospore formation was greatly reduced or absent, suggesting the integration and expression of the genes carried on pMOcosX interfered with fertilization and/or ascospore formation in this homothallic fungus.
Tipo: Journal article
Ano: 2003 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582003000200007
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Quantitative real-time PCR method to detect changes in specific transcript and total RNA amounts Electron. J. Biotechnol.
Baek,Kwang-Hyun; Skinner,Daniel Z.
Quantitative real-time PCR (qRT-PCR), used in conjunction with reverse transcriptase, has been applied to the determination of the number of copies of a transcript per unit mass of RNA, but did not indicate any change in the amount of total RNA per mass of tissue. In the present work, we described a simple method to use qRT-PCR to estimate the change in the amount of total RNA per unit mass of wheat (Triticum aestivum L.) tissue in response to cold temperature. Three qRT-PCR templates, i.e. control, cold-exposed, and one of RNA extracted from a sample consisting of equal masses of control and cold-exposed tissue, were analyzed. The number of copies of target transcript per unit mass of RNA was estimated from the three samples using standard qRT-PCR...
Tipo: Journal article
Ano: 2004 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582004000100008
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