Recombinant Dna & Bio Technology

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Biotechnology

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an industrial process that provides products based on our ability to genetically engineer bacteria or other cells.

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Genetic Engineering

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The process in which foreign genes are inserted into cells. This makes them capable of producing new & different products.

Example: used to make drugs for health, clean up pollutants, alter genotypes (dz resistant plants), etc.

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The result of Genetic Engineering:

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An altered DNA called "Recombinant DNA" (rDNA): DNA from 2 or more different sources.

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Recombinant DNA Vectors:

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To make rDNA you need a vector or a carrier of the foreign DNA, which is to be inserted into the host cell.

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Two types of Recombinant DNA Vectors

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1. Viral DNA

2. Plasmids

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Plasmids

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Small accessory rings of DNA taken from bacteria. These rings carry DNA that are NOT on the bacterial chromosome.

3 Steps:

1) Plasmid is removed from bacterium & the foreign gene is inserted into the plasmid.

2) The altered Plasmid is returned to the bacterium.

3) Bacterium is encouraged to reproduce multiple times. This is referred to as "Gene Cloning". Everytime the bacterium reproduce the new inserted gene is copied.

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Viral DNA

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Viruses are used to carry rDNA in Animal cells. When this virus with its foreign DNA enters the host cell, it releases its DNA.

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Making Recombinant DNA; Two types of Enzymes:

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1. Restriction Enzymes

2. DNA Ligase

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Restriction Enzymes

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An enzyme used to make Recombinant DNA.

Found in bacteria to restrict the growth of viruses within bacteria. There are 100’s of these restriction enzymes that can cleave (cut) DNA at a specific location, sometimes called “molecular scissors”. Foreign DNA can now be placed in this gap. The ends of 2 DNA molecules (foreign and vector) are called "sticky ends" because they ad here by complementary base pairing if cut by same restriction enzyme.

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DNA Ligase

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An Enzyme used to make Recombinant DNA.

Seals any breaks in a DNA molecule and seals foreign DNA into the vector.

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Biotechnology Products

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1. Human Growth Hormones: treats slow growth in kids.

2. Insulin: previously extracted from pancreas of slaughtered pigs, sometimes caused allergic reactions.

3. Tissue Plasminogen Activator (tPA): a protein that is normally only found in small amounts in body. It is very affective in dissolving blood clots. When given to heart attack victims it can greatly reduce the damage caused from a lack of O2.

4. Clotting Factor VIII: treatment of hemophilia.

5. Human Insulin: treatment of diabetes Type I.

6. Bovine growth hormone (bGH): cows produce 25% more milk.

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Making Safer Vaccines

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1. Vaccines that build human immunity can occasionally cause the illness it was intended to prevent. Vaccines produced thru biotechnology don’t.

2. Current recombinant vaccine for hepatitis B.

3. In the future, for Chlamydia, malaria, and AIDS.

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Transgenic Organisms

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Have a foreign gene inserted into them.

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Transgenic Bacteria

(transgenic organism)

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1. Protecting and Enhancing Plants - Used to promote the health of plants, such as to protect against frost damage.

2. Bioremediation - Microbes are used to clean up oil spills.

3. Producing Chemicals - A strain of bacteria has been developed to produce phenylalanine, a chemical needed to make aspartame (NutraSweet).

4. Processing Minerals - Bacteria may help extract copper, uranium, and gold from low-grade sources.

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Transgenic Plants

(transgenic organism)

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Presently there are over 100 genetically engineered plants. It is hoped that one day bioengineered plants will be used for:

A) Increasing plant growth under unfavorable conditions of heat, cold and drought.

B) make seeds & plants more nutritious.

C) develop bruise resistant fruits.

D) produce plants that require less fertilizer.

E) Create plants that produce useful chemicals & drugs.

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Transgenic Animals

(transgenic organism)

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Foreign genes are microinjected into animal eggs.

Gene pharming- the use of transgenic animals to produce pharmaceuticals.

Examples:

1) Bovine growth hormone produce larger fishes, cows, & pigs.

2) The milk of a transgenic sheep contains tPA.

3) A transgenic pig can now produce human hemoglobin.

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Gene Therapy in Humans

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The insertion of genes (DNA) into an individual to treat genetic disorder and other human illnesses.

1. Ex Vivo

2. In Vivo

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Ex Vivo

(Gene Therapy in Humans)

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Cells are removed from a patient, treated, and returned to the patient (ex vivo means living outside the organism).

A retro virus (has RNA genes) is a vector to carry healthy genes into the cells of the patient.

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In Vivo

(Gene Therapy in Humans)

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Cells are not removed from the patient but viruses, laboratory-grown cells, or synthetic carriers are used to introduce genes directly into the patient.

Examples:

1) Adenovirus that contains a gene to treat cystic fibrosis is placed in an aerosol spray for patient use. It is hoped that the modified virus will carry the necessary gene to the trachea, bronchi and pancreas so that the patient will produce normal mucus.

2) Other Diseases being explored: hemophilia, diabetes, Parkinson’s and AIDS.