CATALYST: Gene Expression Review

1. What is RNAi?
2. What is the difference between the control of the lac and trp operon in prokaryotes? (Hint: Look back at biozone pages) 3. How is control of typ operon an example of negative feedback? 4. What are some differences between how gene expression is controlled in prokaryotes and eukaryotes?

Case 1: The Effort to Cure Pituitary Dwarfism

William Harrison and Charles Stratton (Tom Thumb), 1860

Identifying the Problem

Pituitary secretes many hormones, including HGH Gene = GH1

Determining the Mode of Inheritance

Autosomal or sex-linked? Dominant or recessive?

Why did early treatments fail?

A New Plan

Escheria coli

A New Plan
1. Find GH1 2. Obtain copy of the gene 3. Insert gene into E.coli

Using Reverse Transcriptase to Produce cDNAs

Retrovirus life cycle

Using Reverse Transcriptase to Produce cDNAs

DNA Cloning

DNA Cloning = Plasmid =

Cutting and Pasting DNA

Restriction Enzymes (Endonucleases) =

Cutting and Pasting DNA

HG1

__________________= Seals recombinant DNA together

Transformation!

HGH!

HGH!

Fig. 20-7

Screening a DNA Library

TECHNIQUE
Radioactively labeled probe molecules Probe DNA Gene of interest Single-stranded DNA from cell

Multiwell plates holding library clones

Film

Nylon membrane

Nylon Location of DNA with the membrane complementary sequence

DNA Probe =

Mass Producing HGH!

Case 2: Amplification of Fossil DNA The Polymerase Chain Reaction

30,000-year-old Homo neanderthalensis Svante Paabo

Amplifying DNA

Taq Polymerase

DNA Polymerase found in bacterium Thermus aquaticus, first discovered in hot springs of Yellowstone National Park

What did they find out?

30,000-year-old Homo neanderthalensis Svante Paabo 8:45

Oldest DNA Ever Studied

Other Applications of PCR

Case 3: The Development of Golden Rice

Worldwide, about 300 million acres of transgenic crops

Transgenic Plants

Bt corn

Glyphosate resistant soybean Canola, engineered to have less saturated fats

Rice as a Target Crop

250,000 Southeast Asian children go blind each year from Vitamin A deficiency

-carotene

Carotenoids

Synthesizing -carotene in Rice

GGPP

Enzyme 1

Enzyme 2 Phytoene

Enzyme 3 Lycopene

-carotene

Agrobacterium tumefaciens

Ti (tumor inducing) Plasmid

Golden Rice

GMOs
Here you are creating millions of novel organisms, organisms that the Earth has never seen, and then releasing them into the environmentMany of us have spend much of our working lives addressing the terrible ecological problems created by chemical pollution. [But] chemical pollution, however horrible, does dilute over time, and it can often be contained. However, once you release an organism into the environment it cannot be recalled or contained. It will not dilute, but rather will reproduce, disseminate, and mutate. It is unstoppable. -Andrew Kimbrell, Executive Director of the Center for Food Safety