Biotechnology Principles And Processes Part 3

Vectors#

1. Origin of Replication

• DNA sequence where replication begins
• Allows vector to replicate independently in host cell

2. Selectable Markers

• Genes conferring antibiotic resistance or specific traits
• Helps identify and select transformed cells

3. Cloning Sites

• Unique restriction enzyme recognition sequences
• Allow insertion of foreign DNA at specific locations

4. Specialized Vectors

• Purpose-built for specific applications (expression, shuttle)
• Contain additional features for specific host organisms

Host Transformation Methods#

1. Chemical Method

• Uses calcium chloride to make cells competent
• Heat shock enables DNA uptake through cell membrane

2. Micro-injection

• Direct injection of DNA into cell nucleus
• Precise but labor-intensive method

3. Biolistics

• DNA-coated gold particles shot into cells
• Suitable for plant cells with tough cell walls

4. Vector-mediated Transfer

• Uses biological vectors like Agrobacterium
• Natural DNA transfer mechanism into host cells