Cloning 1

IX Gene Transfer Some people’s blood does not clot due to lack of protein IX If they can be supplied  problem solved Production of factor IX is cheapest in a transgenic sheep This is done by associating the human gene which codes for the production of IX with the genetic information for milk production in sheep. Basically: 2

A ewe is treated with fertility drugs to create super-ovulation. Eggs are inseminated. Each fertilized egg has the transgene injected. A surrogate ewe has the egg implanted for gestation. Lambs are born which are transgenic, GMO for this factor IX gene. Each Lamb when mature can produce milk. The factor IX protein is in the milk and so must be isolated and purified before use in human. 3

Gene that codes for protein IX can be inserted in plasmid of host cell (sheep) and plasmid can be inserted in sheep, where protein will be produced Gene that codes for protein IX can be inserted in plasmid of host cell (sheep) and plasmid can be inserted in sheep, where protein will be produced Sheep can synthesize factor IX Sheep can synthesize factor IX The messenger RNA which codes for factor IX is extracted from a human pancreatic cell which produces factor IX. The messenger RNA which codes for factor IX is extracted from a human pancreatic cell which produces factor IX. A selected plasmid is cut using restriction enzymes (endonuclease) which cut the DNA at specific base sequences from the host cell (sheep). A selected plasmid is cut using restriction enzymes (endonuclease) which cut the DNA at specific base sequences from the host cell (sheep). The plasmid from host cell and isolated gene from donor cell are “stuck” together by enzyme DNA ligase. Plasmid is now recombinant and is used as a tool to introduce a new gene in an organism. The plasmid from host cell and isolated gene from donor cell are “stuck” together by enzyme DNA ligase. Plasmid is now recombinant and is used as a tool to introduce a new gene in an organism. Recombinant plasmid is inserted in sheep  sheep produces factor IX found in milk. Recombinant plasmid is inserted in sheep  sheep produces factor IX found in milk. 4 How it is done

5 Advantages / Disadvantages AdvantagesDisadvantages Cheapest way to produce factor XI Incidence rate is 1 / Therefore is it right to genetically modify sheep for such a small sample of population? Production of transgenic sheep by nuclear transfer requires fewer than half the animals needed for pronuclear microinjection Potential danger to sheep

Salt tolerance in tomato plants The new salt-proof tomato isolates salt from the soil, transports it to its leaves and hold it there i.e. away from the fruit The new salt-proof tomato isolates salt from the soil, transports it to its leaves and hold it there i.e. away from the fruit This makes the tomatoes more tolerant to the high levels of salt in the soil (due to modern irrigation techniques) This makes the tomatoes more tolerant to the high levels of salt in the soil (due to modern irrigation techniques) As the tomatoes are more tolerant, it makes it easier to grow in certain regions of high salinity As the tomatoes are more tolerant, it makes it easier to grow in certain regions of high salinity 6

7 How it is done A plasmid is removed from Yeast Hal1 A plasmid is removed from Yeast Hal1 The plasmid is cut open using a restriction endonuclease The plasmid is cut open using a restriction endonuclease The gene to be copied is placed inside the open plasmid The gene to be copied is placed inside the open plasmid The gene is pasted into the plasmid using DNA ligase. The plasmid is now called a recombinant plasmid The gene is pasted into the plasmid using DNA ligase. The plasmid is now called a recombinant plasmid The vector is placed inside the Yeast Hal1 bacterium and the bacterium is given its ideal conditions to grow and proliferate The vector is placed inside the Yeast Hal1 bacterium and the bacterium is given its ideal conditions to grow and proliferate

Advantages / Disadvantages AdvantagesDisadvantages Since there is less damage to the maize crops, there is a higher crop yield which can lessen food shortages. We are not sure of the consequences of humans and animals eating the modified crops. Could be harmful to human as well as animal health. Since there is a higher crop yield, less land is needed to grow more crops. Instead the land can become an area for wild life conservation.  May tackle, to some extent, world hunger Other insects which are not harmful to the crops could be killed. The maize pollen will contain the toxin and so if it is blown onto near by plants it can kill the insects feeding on these plants. There is a reduction in the use of pesticides which are expensive and may be harmful to the environment, wild life and farm workers. Cross pollination can occur which results in some wild plants being genetically modified as they contain new gene. These plants will have an advantage over others as they will be resistant to certain insects and so some plants may become endangered. This will have significant consequences on the population of wild plants. 8