Restriction enzyme

a) Type-I: Cuts everywhere: Eco k and Eco B

b) Type II: Cuts at exact site: EcoRI

c) Type III: Cut near vicinity : EcoP₁ and EcoP₁₅.

Transformation

• Metal size for particle gun method is 0.5-5 µm.
• Particles are called microcarriers.
• Electroporation: Protoplast are subjected to 400 V/cm strength electrical pulses for 10-15 ms.
• Important vector is Agrobacterium tumifaciens.
• T region: Region transferred into host genome.

a) tms1, tms2: Synthesis of IAA

b) tmr: Production of cytokinin

c) Os: Opine synthesis

d) tms1, tms2 and tmr: Tumor production ] oncogenes

• Removal of 3 tumor producing genes from tDNA: disarming
• Autoclaving: Sterlization at 121⁰C at 15 psi for 15-40 min.
• Cybrids: Nuclear gene for one parent and cytoplasm from another parent.
• Cryopreservation: Freezing and storage at -196⁰