Biology 12 Unit 1

DNA Replication The Semi-Conservative Model

Text References: Pg 41-42 (DNA) Pg 114 116 (DNA Replication), Pg 128 137 (Biotechnology)

Deoxyribonucleic Acid (DNA) Chemically, DNA looks a lot like a twisted ladder two long polymers made up of adjoining nucleotides that twist to form a double helix. The 2 sides of the ladder are referred to as the sugar-phosphate backbones The rungs of the ladder are formed when 2 complementary nitrogenous bases are joined by weak hydrogen bonds Adenine (A) always hydrogen bonds to ____________ (complementary bases) Cytosine (C) always hydrogen bonds to ____________ (complementary bases) The sugar of one nucleotide and the phosphate of another are held together by strong phosphodiester bonds

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A Closer Look at DNA DNAs 4 nitrogenous bases (C, G, A, T) can be separated into 2 categories: 1) Purines Have a 2-ring structure E.g. Guanine (G) and Adenine (A) 2) Pyrimidines Have a single-ring structure Ex. Cytosine (C) and Thymine (T) One purine always hydrogen bonds to a pyrimidine. The complementary base pair A T forms _____hydrogen bonds The complementary base pair C G forms _____ hydrogen bonds

Did You Know? If unwound and tied together, the strands of DNA in one cell, would stretch almost six feet long but would only be 50 trillionths of an inch wide. If you uncoil the DNA in all of your cells, you could reach the moon 6000 times! There are 3 billion letters in the human genome and it would take a person who could type 60 words per minute, 8 hours a day, around 50 years to type out the human genome. In 2003, the human genome was completely sequenced, down to the last nucleotide. DNA has a multi-coiled structure that allows an incredible amount of it to be packed into the tiny space within a cells nucleus When cells are not dividing, DNA is called chromatin a loosely coiled tangled mess of DNA inside the nucleus When human cells are preparing to divide, their DNA is tightly coiled into 46 X shaped chromosomes that are arranged in 23 pairs.

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Each chromosome contains sections of DNA called genes Each gene contains a set of instructions to make a specific protein

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DNA Replication As organisms grow their cells divide. DNA replication must occur before cell division so that each new daughter cell has a complete set of genetic information that provides the instructions for all cellular functions. During semi-conservative replication a parent DNA molecule is used as a template to create 2 genetically identical daughter DNA molecules. DNA replication is called semi-conservative because each DNA molecule formed contains one old (parent) strand and one new (daughter) strand. In other words, one of the parental strands is conserved, or present, in each new double helix.

Did You Know? Cancer is characterized by rapidly dividing cells. Treatment for cancer includes chemotherapy drugs that stop replication and therefore cell division. Some chemotherapy drugs are analogs that have a similar, bur not identical, structure to the four nucleotides in DNA. When these are mistakenly used by the cancer cells to synthesize new DNA molecules, replication stops and the cells eventually die.

Steps of Semi-Conservative Replication http://www.youtube.com/watch?v=zdDkiRw1PdU&feature=related

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1) Unzipping A parent DNA molecule unwinds and DNA helicase (an enzyme) breaks the weak hydrogen bonds between complementary base pairs causing the double helix to unzip

Single-strand binding proteins (SSBs) stabilize the unwound DNA parent strands and keep them from joining back together
Direction helicase unzips DNA

helicase SSBs

Replication Fork

Unzipping occurs at multiple sites along a single DNA molecule This allows DNA replication to begin at several points rather than just starting at one end. This greatly speeds up the rate of replication. Remember that in a single DNA molecule there are more than 300 million base pairs to replicate!

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2) Complementary Base Pairing Once the strands have been unwound and separated DNA polymerase (an enzyme) brings in free floating nucleotides to build a new daughter strand by using the parent strand as a template

Daughter Strand Parent Strand

DNA Polymerase

The helix continues to unwind and open allowing the strand grows continuously towards the replication fork.

Old Parent Strands

New Daughter Strand being synthesized by DNA polymerase

The leading strand is the strand that is replicated towards the replication fork The lagging strand is the strand that is replicated in the direction that moves away from the replication fork The lagging strand is replicated in a discontinuous fashion in fragments called Okazaki Fragments

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3) Joining of Adjacent Nucleotides DNA ligase (an enzyme) forms strong phosphodiester bonds between the sugar of one nucleotide and the phosphate group of another nucleotide in each lagging daughter strand. This process continues until two genetically identical DNA molecules are formed. Recombinant DNA (rDNA) - Biotechnology Genetic engineering is a form of biotechnology used to alter the DNA of viruses, bacteria, and other cells for medical, agricultural or industrial purposes. Scientists have the ability to isolate pieces of DNA called genes in one organism and transplant them into the DNA of another organism. The result is an improved organism or an organism which is able to produce a desirable genetic product such as a specific protein it wouldnt otherwise be able to produce. These proteins are then harvested and used in a variety of applications.

Recombinant DNA r(DNA) - contains DNA from two or more different sources
Did you Know? Scientists have created genetically altered pigs that lack a gene involved in rejection of transplanted organs. They could potentially be used to grow organs for human transplants. Did you know that spider silk is the strongest, toughest fibre known to man? Webster & Pete are 2 baby goats that have had the gene from a spider inserted into their genome. This gene causes these goats to produce spider silk in their milk. The silk protein in their milk will be used to make Biosteel a material used to make bulletproof vests, aircraft, & race cars.

Getting vaccines, vitamins and medicines to people in third world countries is very expensive and difficult. Scientists are hoping foods such as bananas could be the answer to this problem. They are using rats genes to develop a vaccine which will be inserted in the genome of the banana.

Scientists have found the gene for green fluorescent protein (GFP) that makes jellyfish glow in the dark. They have inserted this gene into mice, which now glow bright green in the dark!

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Uses of Recombinant DNA:

1. Mass production of proteins is possible and can be produced inexpensively for a wide variety of medical applications. Insulin for diabetics Growth Hormone for Pituitary Dwarfism (low secretion of the hormone) Erythropoietin helps to increase red blood cell production in the bone marrow Tissue Plasminogen Activator enzyme dissolves blood clots in heart attack victims Human gene for Chloride Channel Protein for suffers of cystic fibrosis

2. Safer and cheaper vaccines can be produced Products are safer for human and animal use fewer side affects from contamination or allergic reactions. Cheaper vaccines provide more accessibility to people in underdeveloped countries like those in Africa. Vaccine for hepatitis A, B and C Vaccine for herpes Vaccine for Lyme Disease

3. Ecological Use Transforming bacteria into oil metabolizing bacteria for oil spills.

4. Disease and drought resistant crop production Salmon that grow 10 30 times faster than wild populations Strawberries which are resistant to frost Tomatoes which resist molds and mildews Corn which is drought resistant Maize which produces its own insecticide which kills the stem borer caterpillar By 2003, 81% of all soybeans, 73% of cotton, and 40% of all corn were genetically engineered in an effort to increase crop production.

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How Recombinant DNA is Made

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For example, E. coli is a bacterium that reproduces about every 20 minutes. Segments of DNA can be inserted into E. coli and the bacterium will reproduce those genes too! Protein hormones like insulin can be made by inserting insulin genes into E. coli DNA.

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