Author: Mohammadzadeh, Sara; Khabiri, Alireza; Roohvand, Farzin; Memarnejadian, Arash; Salmanian, Ali Hatef; Ajdary, Soheila; Ehsani, Parastoo
Title: Enhanced-Transient Expression of Hepatitis C Virus Core Protein in Nicotiana tabacum, a Protein With Potential Clinical Applications Document date: 2014_11_24
ID: 3posyr5n_8
Snippet: A. tumefactions strain LB4404 was transformed with PVXcore and pBI-core vectors ( Figure 3 A and 3B) in separate reactions via the standard freeze-thaw protocol (40) . To this end, the HCVcp-coding plant constructs, pBI-core and PVX-core were introduced into Agrobacterium LB4404 (in the case of pBI-core) and into Agrobacterium LB4404; the latter had already harbored the helper plasmid pSOUP (41) (in the case of PVX-core), which is essential for r.....
Document: A. tumefactions strain LB4404 was transformed with PVXcore and pBI-core vectors ( Figure 3 A and 3B) in separate reactions via the standard freeze-thaw protocol (40) . To this end, the HCVcp-coding plant constructs, pBI-core and PVX-core were introduced into Agrobacterium LB4404 (in the case of pBI-core) and into Agrobacterium LB4404; the latter had already harbored the helper plasmid pSOUP (41) (in the case of PVX-core), which is essential for replication of the PVX-GW-based plasmids in Agrobacterium species, respectively. Subsequently, transformed Agrobacterium cells were selected on plates containing 100 µg/ mL rifampicin (RIF) and 50 µg/mL kanamycin (KAN) and incubated for 72 hours at 28°C. The transformed colonies were confirmed by gene-specific colony PCR using F-Kozak-VX and R-core-VX-specific primers, as described above. All the chemicals, culture media and antibiotics were purchased from Sigma (US) and Merck (Germany) companies.
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