Plant genetic engineering has been considered to be a solution for such global challenges. Molecular biology technology using synthetic or modified genes with appropriate sensible genetic engineering strategies can help to increasing or create a new product. This potentially would lead to high production yields at low costs of better quality oils which are important for green energy and industrial materials.
2. Clip main shoots to encourage proliferation of many secondary branches.
3. Waiting until plants have flowers
4. Prepare Agrobacterium tumefaciens strain carrying gene of interest on a binary vector. Grow a large liquid culture @ 28-30oC in LB with antibiotics to select for the binary plasmid, or grow in other media.
5. Spin down Agrobacterium, resuspend to OD600 = 0.8 (can be higher or lower) in 5% Sucrose solution (made fresh is better, no need to autoclave). Add Silwet L-77 to a concentration of 0.05% (500 ul/L) and mix well. You will need 400-500 ml for 6-9 3.5" (9cm) pots.
6. Dip flower parts of plant in Agrobacterium solution for 2 to 3 minutes in vaccum.
7. Place dipped plants laid down on a tray and cover for 16 to 24 hours to maintain high humidity.
8. Tie plants in the pot with a stick as long as plant high. Water and grow plants normally. Stop watering as seeds become mature. Do not water on the top part (seed and flower)
9. Harvest dry seed. Transformants are usually all independent, but are guaranteed to be independent if they come off of separate plants.
10. Select for transformants using fluorescence or herbicide (basta) selectable marker.
11. Put the transgenic seed (color) to soil for next generation.
12. Look at transgenic seed with red color
Transgenic seeds (left) and WT seeds (right). It was took in regular light and with out any filter by Kodak camera
Video clip for Camelina transformation
References:
Bechtold, N., Ellis, J., and Pelletier, G. (1993). In planta Agrobacterium-mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C. R. Acad. Sci. Paris, Life Sciences 316:1194-1199.
Last Updated on Wednesday, 24 February 2010 21:28
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3:1 non green and green seed (Zs-green gene)
3:1 non green and green seed (Zs-green gene)
BNL Biology Department
Biodiesel become clear after washing a couple of days in room temprature
Biodiesel from camelina seed (top layer) before washing with water
Biodiesel is washing with waters
Camelina plants selected by basta after 10 days
Camelina plants selecting by basta 1 week
Camelina_oil
Camelina_oil_2818_1
EXPRESSION OF THE Ds-Red GENE IN THE TRANSGENICPROTOCORM-LIKE BODY OF THE Dendrobium