Renno D V, Saunders G, Bull A T, Holt G
School of Biotechnology, Polytechnic of Central London, UK.
Appl Microbiol Biotechnol. 1990 Dec;34(3):364-7. doi: 10.1007/BF00170061.
A number of transformants of Penicillium chrysogenum have been obtained with the plasmid vector p3SR2. Southern analysis showed that transformation had occurred by integration of vector sequences into the nuclear DNA of the fungus. A number of transformants were tested for stability of the transformed phenotype in agar medium and some were found to be unstable. Two transformants, shown to be stable in agar culture, were grown in 5-1 batch fermentors for further stability tests. Over periods of up to 312 h in the fermentor both transformants were 100% stable with respect to the transformed phenotype. In addition Southern analysis of DNA extracted from the spent mycelium showed that no change had occurred in the position of the integrated vector sequences within the transformant nuclear DNA.
利用质粒载体p3SR2已获得了许多产黄青霉转化体。Southern分析表明,转化是通过载体序列整合到真菌的核DNA中而发生的。对一些转化体在琼脂培养基中进行转化表型稳定性测试,发现其中一些是不稳定的。有两个在琼脂培养中表现稳定的转化体,在5-1分批发酵罐中培养以进行进一步的稳定性测试。在发酵罐中长达312小时的时间段内,这两个转化体的转化表型均100%稳定。此外,对从用过的菌丝体中提取的DNA进行Southern分析表明,转化体核DNA中整合的载体序列位置没有发生变化。