Mort-Bontemps M, Fèvre M
Laboratoire de Biologie Cellulaire Fongique, CGMC UMR CNRS 5534, Université Lyon 1, 43, Boulevard 11 Novembre 1918, F-69622 Villeurbanne Cedex, France.
Curr Genet. 1997 Mar;31(3):272-5. doi: 10.1007/s002940050205.
Saprolegnia monoïca was stably transformed to hygromycin resistance using the plasmids pTH210 and pHAM34H containing a bacterial phosphotransferase gene fused to regulatory sequences from genes of another Oomycete. Vectors pBT6, pCM54, used to transform higher fungi, yielded no stable transformants. DNA hybridizations indicated that transformation resulted from a single-copy integration of the transforming vector. Development of non-resistant subcultures from transformed colonies revealed that the transgene could become silent and was not uniformly expressed in the transformants.
使用含有与另一种卵菌基因的调控序列融合的细菌磷酸转移酶基因的质粒pTH210和pHAM34H,将单孢水霉稳定转化为对潮霉素具有抗性。用于转化高等真菌的载体pBT6、pCM54未产生稳定的转化体。DNA杂交表明,转化是由转化载体的单拷贝整合引起的。从转化菌落中产生无抗性的继代培养物表明,转基因可能会沉默,并且在转化体中并非均匀表达。