BiotechnologyEllie Wortman

Zanib Sarfraz

What is biotechnology?

• Living organisms, or their products, modify human life and the environment

• Began nearly 10,000 years ago• Manipulate conditions to yield more end product• From nomadic hunter to the settled farmer

– Crops were essential– Increase yield – Improve taste

• This was only the beginning of biotechnology as it would become more complex

Where is it Used?

• Agriculture– Produce plants that are more resistant

• Bioremediation• Food Processing• Energy Production• Major Subjects

– Chemistry– Biology– Physics

• Three Major Branches– Genetic Engineering– Diagnostic Techniques– Cell/Tissue Techniques

Biotechnology and viruses and bacteria• Recombinant DNA – DNA from two different sources

– Transgenic • Biotechnology puts certain genes in viruses and bacteria• Viruses are non living

• VIRUS STRUCTURE -Nucleic acid surrounded by a capsid coat (made of proteins)

Recombinant DNA and Cloning

• DNA created artificially• the recombinant molecule must be replicated many times to provide

material for analysis• recombinant DNA must be taken up by the cell in a form in which it can be

replicated and expressed. – vectors

• Cloning– Producing many identical copies of the same recombinant molecule– Sticky ends– Two samples of DNA are mixed and some fragments of one organism

will stick to the other

Genomic Libraries

• Bacteria and virus vectors– Fragments

• Collection of bacteria or viruses is called a genomic library• Finding Genes in a Gene Library

– Microscope– Autoradiography– Radioactive probes

• Radioactive or fluorescent helps to visualize the gene• disease-causing microorganisms• defective (disease) genes• various cancers

PCR

• The polymerase chain reaction • Used to make many copies of small pieces of DNA • Because techniques in biotechnology usually require many copies of

genes, PCR has allowed much of the biotechnology development that we have seen in recent years

• Materials Needed:– Primers– DNA Polymerase– Nucleotides

PCR Procedure

1. The DNA in question is heated to separate the two strands of the double helix.

PCR Procedure

2. After the strands are separated, the DNA is cooled and the primers attach.

PCR Procedure

3. Next, DNA polymerase attaches and copies the strand.

PCR Procedure

4. The solution is then heated and cooled again as described above at regular intervals. Each time it is cycled through this heating and cooling procedure, the DNA replication process repeats itself and the amount of DNA produced is doubled.

DNA Fingerprinting (RFLP Analysis)• The DNA of an organism is cut up into

fragments using restriction enzymes• A large number of short fragments of DNA

will be produced• Restriction enzymes always cut at the same

base sequence• Because no two people have identical DNA,

no two people will have the same length fragments

Gel Electrophoresis

• Electrophoresis is a technique used to separate the DNA fragments according to their size. They are placed on a sheet of gelatin and an electric current is applied to the sheet. DNA is charged and will move in an electric field toward the positive pole.

Gel Electrophoresis

• Smallest fragments will move the fastest because they are able to move through the pores in the gelatin faster

• Bands will be produced on the gelatin where the fragments accumulate• The shortest fragments will accumulate near one end of the gelatin and

the longer, slower-moving ones will remain near the other end

Gel Electrophoresis Uses

• This procedure requires a large amount of DNA so it is often used in conjunction with PCR – uses are identification of diseased genes including oncogenes,

identification of viral infections, determining family relationships among individuals, and identifying tissue found at a crime scene

• Sickle Cell disease, Huntington’s disease, Duchenne muscular dystrophy• Taxonomists can use this technique to explore evolutionary relationships• The procedure for sequencing and mapping DNA requires RFLP analysis

Biotechnology in Human Therapy• Tissue plasminogen activator(TPA) for dissolving blood clots • adenosine deaminase (ADA) for treating some forms severe combined

immunodeficiency(SCID) • Parathyroid hormone• several conoclonal antibodies• Behaptitis B surface antigen(HBsAg) to vaccinate against the hepatitus B

virus• C1 inhibitor(C1INH) used to treat hereditary angioedema• Granulocyte-macrophage colony-stimulating factor (GM-CSF) for

stimulating the bone marrow after a bone marrow transplant • Granulocyte colony-stimulating factor(G-CSF) for stimulating neutrophil

production, e.g., after chemotherapy and for mobilizing hematopoietic stem cells from the bone marrow into the blood.

Crossword Puzzle Answer Key

CURRENT CLUES ACROSS: 5 MANY |Eukaryotic cells have _____ thousands of genes 6 PLASMIDS |Smal, accessory rings of DNA 9 VIRUSES |The vectors of choice for inserting genes into animal cells 12 RADIOACTIVEPROBES |Can be used to find colonies that have specific genes 13 VECTORS |Pieces of DNA that are used to transfer genes into a host cell 14 INTRONS |Eukaryotic genes contain _____ 15 BIOTECHNOLOGY |Refers to technology used to manipulate DNA 16 GENOME |A ______ is all the genes in a particular organism CURRENT CLUES DOWN: 1 POLYMERASECHAINREACTION |Can be used to make many copies of small pieces of DNA 2 RECOMBINANTDNA |Refers to DNA from two different sources 3 PROBES |Short, single-stranded segments of DNA whose base sequence matches part of the gene is question 4 RNA |The genetic material for retroviruses 7 STICKYENDS |Fragments of DNA that has been cut with restriction enzymes have unpaired nucleotides at the ends called _____ 8 RESTRICTION |Enzymes used as a defense mechanism to cut up the DNA of viruses or other bacteria 10 AIDSVIRUS |An example of a retrovirus 11 LIGASE |DNA _____ is used to seal the fragments between sticky ends

Brown, C (1994). Recombinant DNA. Kimball’s Biology Pages. Retrieved fromhttp://users.rcn.com/jkimball.ma.ultranet/BiologyPages/R/RecombinntDNA.html

Michael J. Gregory, Ph.D (2006). Biotechnology. The Biology World. Retrievedfrom http://faculty.clintoncc.suny.edu/faculty/michael.gregory/

files/Bio%20 00/Bio%20100%20Lectures/Biotechnology/biotechn.htmNational Health Mueseum (1990). Where did Biology Begin? Access Excellence Resource Center. Retrieved fromhttp://www.accessexcellence.org/RC/AB/BC/Where_Biotechnology_gin.phpPicture Slide 4 retrievedhttp://learn.genetics.utah.edu/archive/sars/images/virus_structure.jpg