Duva’s Reading Guide for Chapter 13: Biotechnology

Concept 13.1: Recombinant DNA Can Be Made in the Laboratory

I. Recombinant DNA -A. Restriction enzymes

1. Origin

2. Function

B. Recognition sequence/restriction site –

1. Blunt –

2. Sticky –

a. BamHI

b. HindIII

c. EcoRI

3. Palindrome

C. Modification of bacterial DNA

D. Restriction digest

II. Gel electrophoresis separates DNA fragmentsA. Gel electrophoresis

1. Purpose

2. Design

a. Gel

b. Buffer

c. Electric field

B. How does it work?

C. Information obtained from the DNA sample

1. Number of fragments

2. Sizes of fragments

3. Relative abundance of a fragment

III. Recombinant DNA can be made from DNA fragmentsA. DNA ligase

B. Cohen and Boyer

1. Bacterial plasmid

2. Results

Concept 13.2: DNA Can Genetically Transform Cells and Organisms

I. Clone

II. Transformation

III. Transgenic

IV. Selectable marker

V. Model organisms for researchA. Bacteria

1. Pros

2. Cons

B. Yeasts

1. Pros

2. Cons

C. Plant cells

1. Pros

2. Cons

D. Cultured animal cells

1. Pros

2. Cons

VI. Methods for inserting DNA into host cellsA. Chemical treatment

B. Electroporation

C. Use of viruses and bacteria as vectors

D. Inject DNA into nuclei of fertilized eggs

E. “gene guns”

VII. Replicon – A. Can be inserted into host chromosome randomly

B. Vector

1. Plasmids

a. Small

b. Restriction enzyme recognition sites

c. Selectable marker (antibiotic resistance)

d. Replicate independently of host chromosome

2. Plasmid vectors for plants

a. Agrobacterium tumefaciens

b. Plasmid name

c. How does it work?

3. Viruses

a. Bacteriophage λ

b. Pros

C. Reporter genes

Concept 13.3: Genes and Gene Expression Can Be Manipulated

I. Sources of DNA fragmentsA. Genomic library

B. cDNA libraries

C. synthetic DNA and PCR

II. DNA mutations can be made in the lab

III. Genes can be inactivated by homologous recombinationA. Inactivate genes to understand their function

B. Stem cells

Concept 13.4: Biotechnology Has Wide Applications

I. Protein factoriesA. Expression vectors

1. Prokaryotes

2. Eukaryotes

II. Medically useful proteinsA. Human insulin

1. Engineered plasmid

2. Bacteria factories

3. Chemical treatment

B. Pharming1. Protein produced in milk

2. HGH

III. Agriculture – transgenic plantsA. Produce insecticides

B. Improved nutrition

C. Adapt to environment

IV. Public concernA. Unnatural

B. Unsafe

C. Dangerous to the environment