TY  - GEN
T1  - An updated pGREEN-based vector suite for cost-effective cloning in plant molecular biology
AU  - Pratt, Anna I.
AU  - Knoblauch, Jan
AU  - Kunz, Hans-Henning
DO  - 10.17912/micropub.biology.000317
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000317/
AB  - Traditional binary shuttle vectors used for creating plant T-DNA insertion mutants can be difficult to work with for several reasons. First, their large size (> 10 kB) and low-copy origin of replication result in low plasmid yields under typical growth conditions. This makes time consuming, and costly, midi or maxi scale plasmid isolation a requirement. Second, accurate sequence information is often not available, rendering vector modification difficult. Finally, cloning techniques that are reliant on restriction enzyme sites prohibit deliberate design of vectors resulting in haphazard configuration of components. Newer cloning techniques, such as Gibson (Gibson et al., 2009) and In-FusionTM assembly, offer unlimited flexibility but rely on sufficient vector fragment quantity; this can be achieved either by restriction digest or by PCR. However, the large size and low DNA yield of binary shuttle vectors make their use in assembly reactions challenging; this is especially true for large batch DNA library cloning.
PY  - 2020
JO  - microPublication Biology
ER  -