pLDutr-CTB-Fx3Pris

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Bombardment was done using the Bio-Rad PDS-1000/He gene gun as described previously (Verma et al., 2008). Sterile tobacco leaves were placed abaxial side up, whereas sterile lettuce leaves were placed abaxial side down on Murashige and Skoog (MS) medium and were bombarded under sterile conditions using pLDutr-CTB-Fx3Pris or pLsLF-CTB-Fx3Pris coated with gold particles prepared according to (Kumar and Daniell, 2004). Bombarded leaf pieces were incubated in the dark for 2 days at room temperature, and then leaf segments were transferred to regeneration media of plants (RMOP) containing 500 mg/L spectinomycin for tobacco or regeneration media of lettuce (RMOL) containing 100 mg/L spectinomycin (Ruhlman et al., 2010).

Protocol tips

Bombardment was done using the Bio-Rad PDS-1000/He gene gun as described previously (Verma et al., 2008). Sterile tobacco leaves were placed abaxial side up, whereas sterile lettuce leaves were placed abaxial side down on Murashige and Skoog (MS) medium and were bombarded under sterile conditions using pLDutr-CTB-Fx3Pris or pLsLF-CTB-Fx3Pris coated with gold particles prepared according to (Kumar and Daniell, 2004). Bombarded leaf pieces were incubated in the dark for 2 days at room temperature, and then leaf segments were transferred to regeneration media of plants (RMOP) containing 500 mg/L spectinomycin for tobacco or regeneration media of lettuce (RMOL) containing 100 mg/L spectinomycin (Ruhlman et al., 2010).

Publication protocol

Bombardment was done using the Bio-Rad PDS-1000/He gene gun as described previously (Verma et al., 2008). Sterile tobacco leaves were placed abaxial side up, whereas sterile lettuce leaves were placed abaxial side down on Murashige and Skoog (MS) medium and were bombarded under sterile conditions using pLDutr-CTB-Fx3Pris or pLsLF-CTB-Fx3Pris coated with gold particles prepared according to (Kumar and Daniell, 2004). Bombarded leaf pieces were incubated in the dark for 2 days at room temperature, and then leaf segments were transferred to regeneration media of plants (RMOP) containing 500 mg/L spectinomycin for tobacco or regeneration media of lettuce (RMOL) containing 100 mg/L spectinomycin (Ruhlman et al., 2010). The appearance of green shoots was seen after 4– 6 weeks. These shoots were allowed to grow, and the leaves were cut and subsequently transferred to RMOP or RMOL containing 500 mg/L or 100 mg/L spectinomycin, respectively, for the second round of selection. After new shoots formed, these were transferred for the third round of selection to MS containing 500 mg/L spectinomycin for tobacco or 100 mg/L spectinomycin for lettuce and allowed to establish roots. Plants were then transferred to the greenhouse.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Eukaryotic cells - lettuce leaves proinsulin using pLDutr-CTB-Fx3Pris from Henry Daniell, Department of Biochemistry, School of Dental Medi.

Manufacturer protocol

Download the product protocol from Henry Daniell, Department of Biochemistry, School of Dental Medi for pLDutr-CTB-Fx3Pris below.

Videos

Check out videos that might be relevant for performing Protein Expression Eukaryotic cells - lettuce leaves proinsulin using pLDutr-CTB-Fx3Pris from Henry Daniell, Department of Biochemistry, School of Dental Medi. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.