Transformation of csp Gene into Tobacco Plant Mediated by Agrobacterium tumefaciens
No Thumbnail Available
Date
2022-01-19
Journal Title
Journal ISSN
Volume Title
Publisher
Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract
Abstract. At the cellular level, heat stress makes RNA misfolding, so protein synthesis cannot occur. The cold shock protein (csp) gene is a type of RNA chaperon from bacteria proven to be successful in protecting bacterial RNA from misfolding. This protein is naturally abundant in the nucleus, cytoplasm, and endoplasmic reticulum (ER). To insert
genes from bacteria, Agrobacterium tumefaciens is needed as an intermediary to insert the genes into tobacco models. Previously, we succeed transfer plasmid pCAMBIA1300int with carrier csp gene into vector A. tumefaciens and already into transformation tobacco. This study aimed to obtain transgenic tobacco containing the csp gene. This study used
Nicotiana tobaccum L. cv. Samsun leaf pieces inoculated for 20 minutes with A. tumefaciens strain LBA4404 carrying plasmid pCAMBIA1300int-csp. The transformants were selected on medium with 20 mg/l hygromycin, then continued with 50 mg/l to get regenerated transformants. A. tumefaciens mediated tobacco transformation produced 65% efficiency. PCR analysis indicated plantlets which passed the selection medium showed 100% of the csp gene containment. Chisquare analysis on the hpt gene showed one copy/plant with a ratio (3:1) when tobacco transgenic plants were cultivated
on medium containing hygromycin. Further studies are needed regarding the field test of this plant.
Description
Keywords
Tobacco, Agrobacterium tumefaciens, transgenic