Chapter 10 Genetic Engineering

Recombinant DNA technology

remove and combine gene into a different organism

A cloning host is usually a _____ or _____ that can ____ a gene and then ____ it into a protein.

bacterium, yeast, replicate, translate

What is the first step of recombinant DNA technology?

Gene inserted into plasmid

What is the second step of recombinant DNA technology?

Plasmid put into bacteria cell

What is the third step of recombinant DNA technology?

Host cell grown in culture

What is the fourth step of recombinant DNA technology?

Research and applications

What is involved in the first step of recombinant DNA technology?

restriction endonuclease, ligase

What is involved in the second step of recombinant DNA technology?

artificial transformation, competency

Restriction endonuclease

breaks bonds in DNA, creates sticky ends

What is the purpose of sticky ends?

join complementary DNA from other organisms

Ligase

seals plasmids back together

Artificial transformation

lab conditions that allow bacteria to take in DNA easily

Competence

ability of genes to cross plasma membrane

How do you make a gene more competent? 

calcium chloride and heat shock

Transgenic plants

have DNA from other organism

BT gene

produce toxin to protect from caterpillars

Glyphosate resistance gene 

protects food plants from specific herbicide 

Glyphosate 

herbicide kills weeds 

What is shown in these images?

Argobacterium tumefaciens

Argobacterum tumefaciens causes what disease?

Crown-gall

Argobacterium tumefaciens is identified by what? What is this condition caused by?

tumor like growth, plasmid insertion 

DNA profiling

specific DNA pattern obtained from person or sample

What are some examples of DNA profiling? 

forensics, paternity, identifying human remains

List the 6 step procedure of DNA profiling.

collect sample, isolate DNA, cut fragments, replicate, separate and stain, analyze

Remember DNA profiling by:

CIC! RSA!

You can amplify DNA by PCR, which stands for ___ ____ ____. This _____ a segment of DNA across several orders of magnitude, generating thousands to millions of ____ of a particular ____ sequence. 

polymerase chain reaction, amplifies, copies, DNA 

How many steps are in the polymerase chain reaction cycle?

3

What are the steps of polymerase chain reaction?

Denaturation, priming, extending

In denaturation of PCR the DNA strands will _____, which causes what?

separate, exposes genes 

Priming

DNA primers attach to 3’ end

During extending, the DNA polymerase is ____ ____ and extends the primers in a ? to ? direction.

heat stable, 5, 3

What is occurring in this step?

Denaturation 

What is occurring in this step?

Priming 

What is occurring in this step?

Extension 

Thermocycler 

amplifies DNA segments via PCR 

What is this?

Thermocycler

Gel electrophoresis

separates DNA based on size

What is this device used for?

Gel electrophoresis 

Where is the DNA sample placed in gel electrophoresis?

one end

In which direction do DNA molecules move in gel electrophoresis due to the current?

from negative to positive

What kind of fragments move the easiest through the gel matrix in electrophoresis?

shortest

DNA fragments appears as ____, each one is a collection of DNA molecules of the ____ length.

bands, same

The location of a band tells you what? 

Fragment length 

The width of a band tells you what?

Abundance

The shortest fragments travel the _____, the widest bands are the most ____.

fastest, abundant