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Direct DNA transfer to plants
- 1. PCTE Group of Institutes, DIRECT DNA TRANSFER TO PLANTS 4/24/2022 Faculty of Pharmaceutical Sciences, PCTE Group of Institutes 1 PRESENTED TO PRESENTED BY Ms. NEHA HARSHDEEP SINGH ASSISTANT PROFESSOR ROLL NO. 2021481 FACULTY OF BIOTECHNOLOGY 4th SEMESTER
- 2. CONTENTS :- • Introduction. • Different types of methods. •Natural methods •Physical methods •Chemical methods • Applications. • Conclusion • References 4/24/2022 PCTE GROUP OF INSTITUTES 2
- 3. 4/24/2022 PCTE GROUP OF INSTITUTES 3 Introduction • Transfer a gene from one DNA molecule to another DNA molecule. • Transgene and the organism that develop after a successful gene transfer is known as transgenic.
- 4. 4/24/2022 PCTE GROUP OF INSTITUTES 4 BY NATURAL METHODS 1. CONJUGATION • Presence of a special plasmid called the F plasmid. • F+ plasmid is integrated within the bacterial chromosome, the cell is called an H fr cell (high frequency of recombination cell).
- 5. 4/24/2022 PCTE GROUP OF INSTITUTES 5 2. TRANSFORMATION • Transformation is the direct uptake of exogenous DNA from its surroundings and taken up through the cell membrane. • This is commonly done using calcium chloride which parabolizes the cell membrane.
- 6. 4/24/2022 PCTE GROUP OF INSTITUTES 6 1. ELECTROPORATION BY PHYSICAL METHODS • Electroporation uses electrical pulse to produce transient pores in the plasma membrane thereby allowing DNA into the cells. • Electroporation as a transformation method is fast, convenient, simple, and inexpensive and has low cell toxicity.
- 7. 4/24/2022 PCTE GROUP OF INSTITUTES 7 2. MICROINJECTION • Microinjection is the process of transferring the desirable DNA into the living cell ,through the use of glass micropipette. • Microinjection involves direct mechanical introduction of DNA into the nucleus or cytoplasm using a glass micro capillary injection pipette.
- 8. 4/24/2022 PCTE GROUP OF INSTITUTES 8 3. PARTICLE BOMBARDMENT GUN METHOD (BIOLISTICS): • Effective method for gene transfer, and creation of transgenic plants. • In this method, 1-2 µm gold or tungsten particles coated with DNA are shot into the plant cells using a helium pressure particle gun device.
- 9. 4/24/2022 PCTE GROUP OF INSTITUTES 9 BY CHEMICAL METHODS 1. CALCIUM PHOSPHATE MEDIATED DNA TRANSFER • 3 phosphate facilitates the binding of the condensed DNA in the co-precipitate to the cell surface, and the DNA enters the cell by endocytosis.
- 10. 2. POLYETHYLENE GLYCOL MEDIATED TRANSFECTION 4/24/2022 PCTE GROUP OF INSTITUTES 10 • This method is utilized for protoplast only. • This method is not perfect because many treated cells do not contain any transfer DNA. • Sometimes the foreign DNA is degraded in the cytoplasm before reaching the nucleus.
- 11. 4/24/2022 PCTE GROUP OF INSTITUTES 11 APPLICATIONS • Clinical gene transfer applications • Vaccine Development • Production of transgenic animals • Treatment of Cancer, AIDS • Gene Discovery • Gene Therapy 2 • Enhancing the resistance of plants • GMO ( Genetically Modified Organisms)
- 12. 4/24/2022 PCTE GROUP OF INSTITUTES 12
- 13. 4/24/2022 PCTE GROUP OF INSTITUTES 13 REFERENCES USED FOR RESEARCH: • Fraley RT, Rogers SG, Horsch RB, Sanders PR, Flick JS, Adams SP, Bittner ML, Brand LA, Fink CL, Fry JS, et al. Expression of bacterial genes in plant cells. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4803–4807. • Plant Cell Rep;6(4):297-301 – PubMed • Plant Mol Biol;5(4):235-46 – PubMed • Horsch RB, Fraley RT, Rogers SG, Sanders PR, Lloyd A, Hoffmann N. Inheritance of functional foreign genes in plants. Science. 1984 Feb 3;223(4635):496–498.
- 14. 4/24/2022 PCTE GROUP OF INSTITUTES 14