Protoplast transformation

Agrobacterium-mediated stable transformation -Plant Agrobacterium-mediated stable transformation -Cell culture Transient protoplast transformation Particle bombardment-based transient expression Agrobacterium- mediated transient expression... 

Salmenkallio, M., Hannus, R., Teeri, T.H. Kauppinen, V. (1990). Regulation of a-amylase promoter by gibberellic acid and abscisic acid in barley protoplasts transformed by electroporation. Plant Cell Reports 9, 352-5. 

A number of different methods for stable or transient genetic transformation of plants or plant cells have been developed [13-15]. These comprise particle bombardment, Agro actermm-mediated transformation, floral dip transformation, agrodrench, viral vectors, protoplast transformation and ultrasound. These are the main techniques for the genetic transformation of plants, and many of them have also been applied for the transformation of secondary metabolite pathways in an attempt to alter the metabolic pathways of target... 

Campoy, S., Perez, F., Martin, J. F., Gutierrez, S., and Liras, P. 2003. Stable transformants of the azaphilone pigment-producing Monascus purpureus obtained by protoplast transformation and Agrobacterium-mediated DNA transfer. Curr. Genet. 43(6), 447-452. 

Katsumata, R., Ozaki, A, Oka, X, and Furuya, A. (1984) Protoplast transformation of glutamate-producing bacteria with plasmid DNA. / Bacte-riol, 159 (1), 306-311. 

A classical way to transform noncompetent Bacilli with replicative plasmids is protoplast transformation. A detailed protocol for protoplast transformation has been documented in the handbook Molecular biological methods for BacUlus by Harwood and Cutting [116]. 

Figure 7.3 "Crossbreeding" by iysed protoplast transformation. (1) A competent B. subtilis cell with wild-type alleles (black dots) is mixed in a hypotonic buffer with a 6. subtilis protoplast comprising a multitude of mutant alleles (black triangles). The competent cell takes up the chromosome of the burst protoplast. CSM is a counterselection marker on the chromosome of the competent wild-type...

The fourth method is to modify DNA against host restriction in vitro. Matsushima and Baltz (21) have demonstrated that cell-free extracts of SoceWopotyspora spinosa can be used to modify plasmid DNA to allow successful protoplast transformation in this highly restricting strain. So far, this method has been limited to a single plasmid. McCue ei al. (31) have modified plasmid DNA with a combination of SssI methylase, which methylates CG sequences at the C residue, and Alul methylase, which methylates AG(Tr sequences at the C residue. The modified plasmids were successfully introduced into Streptomyces griseus by protoplast transformation transformations were not otherwise successful because of the strong restriction harriers. 

Other than conditions specified above, the protoplast transformation coiulitions described by Lampel and Strohl (275) are still followed in our laboratory for most anthra-cycline-producing strains. No technique has yet been published for electroporation of anthracycline-producing streptomycetes. 

---