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Recombining ofDNAmolecules from two different species that are inserted into a host organism to produce newgeneticcombinations that are

of value to science, medicine,agriculture, orindustry. Using this technology, scientists are able to isolate agene, determine

itsnucleotidesequence, study its transcripts, mutate it in highly specific ways, and reinsert the modified sequence into a living organism. The

processes of DNAcloningandsequencingare used to compare different organisms for evolutionary relatedness and to determine gene

function. Recombinant DNA technology can also be used to studymutations and their biological effects, such as the role of specific mutations

in disease or abnormal drugresponse. Other applications of recombinant DNA technology includegene therapy, reverse genetics, diagnostics,

genomics, andproteinmanufacture (the preparation of large amounts of protein for basic research or medicinal use, such as commercially

producedinsulin).genetic engineering;biotechnology.

Read more: http://www.answers.com/topic/recombinant-dna-technology-1#ixzz1xowTYVdk

How is Recombinant DNA made?

There are three different methods by which Recombinant DNA is made. They are

Transformation, Phage Introduction, and Non-Bacterial Transformation. Each

are described separately below.

Transformation

The first step in transformation is to select a piece of DNA to be inserted

into a vector. The second step is to cut that piece of DNA with a restriction

enzyme and then ligate the DNA insert into the vector with DNA Ligase. The insert contains a selectable

marker which allows for identification of recombinant molecules. An antibiotic

marker is often used so a host cell without a vector dies when exposed to a certain

antibiotic, and the host with the vector will live because it is resistant.

The vector is inserted into a host cell, in a process called transformation. One

example of a possible host cell is E. Coli. The host cells must be specially

prepared to take up the foreign DNA.

Selectable markers can be for antibiotic resistance, color changes, or any other

characteristic which can distinguish transformed hosts from untransformed hosts.

Different vectors have different properties to make them suitable to different

applications. Some properties can include symmetrical cloning sites, size, and

high copy number.

Non-Bacterial Transformation

This is a process very similar to Transformation, which was described above. The

only difference between the two is non-bacterial does not use bacteria such as E. Coli

for the host.

In microinjection, the DNA is injected directly into the nucleus of the cell being

transformed. In biolistics, the host cells are bombarded with high velocity

microprojectiles, such as particles of gold or tungsten that have been coated

with DNA.

Phage Introduction

Phage introduction is the process of transfection, which is equivalent to transformation,

except a phage is used instead of bacteria. In vitro packagings of a vector is used.

This uses lambda or MI3 phages to produce phage plaques which contain recombinants.

The recombinants that are created can be identified by differences in the

recombinants and non-recombinants using various selection methods.

Why is rDNA important?

Recombinant DNA has been gaining in importance over the last few years, andrecombinant DNA will only become more important in the 21st century as genetic

diseases become more prevelant and agricultural area is reduced. Below aresome of the areas where Recombinant DNA will have an impact.

Better Crops (drought & heat resistance) Recombinant Vaccines (ie. Hepatitis B) Prevention and cure of sickle cell anemia Prevention and cure of cystic fibrosis Production of clotting factors

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Production of insulin Production of recombinant pharmaceuticals Plants that produce their own insecticides Germ line and somatic gene therapy

What does the future hold?Now that we've figured out the basics behind what Recombinant DNA are, it's

time to look at how Recombinant DNA will impact the future. Which industriesand fields will be shaped by rDNA? How will rDNA effect the health andlifestyles of RPI students in the next generation?

Impacts of Rdna

Good

Improved Medicines Improved Livestock (resistance to disease) Improved Crops (resistance to disease, higher yields) Prevention of Genetic Diseases Lowering the cost of medicines (i.e. Insulin) Safer Medicines (i.e. Insulin) Treatment for pre-existing conditions (i.e. Cancer)

Bad

Safety concerns (viruses developing antibiotic resistance) Environmental concerns (developing resistance to fungi) Ethical dilemmas over human treatment (i.e. are we playing God?) Potential for Experimental abus (doctors using patients as test subjects) Germline treatment going from treating diseases to a method for

picking the traits you want in a child (i.e. specifying hair and eye color)

Were it not for recombinant DNA (rDNA) technology, also referred to as gene splicing,cloningwould merely be aninteresting, but useless side path ofmolecular biology. The opposite is also true: without cloning, recombinant DNA would

continue to be an expensive and time-consuming process. The reason for this is simple. Recombinant DNA is difficult and has alow success rate. When it is achieved, there is no guaranteed way to pass the genetically altered organism's newly obtainedtrait on to subsequent generations. However, a organism can be cloned, thus producing thousands of organisms with thedesired trait.

The first task of the gene splicing process is to isolate thegenethat is to be inserted into theDNA. For this example, ahumanproteinwill be inserted into another animals DNA, just as in the case ofPolly. This can be done through the lengthyprocess of searching through the entire genome. In the case of humans, the genome is being mapped by the Human GenomeProject. However this is costly and has not yet been completed. Another way is to isolate the RNAwhich is producing theprotein from the specific area of the body. This RNA can then be converted into DNA using reverse transcriptase,anenzymeused by retroviruses to convert their RNA genes into DNA.

Recombinant DNA is made possible by two important enzymes. Restriction enzymesandDNA ligaseare the two principaltools, first used byPaul Bergin1972, employed to alter DNA. Restriction enzymes are used to "cut" DNA at a specificlocation. These are used if the DNA must be removed from the entire strand of DNA (as in the first method for gene isolationabove) and also to open the section of DNA into which the isolated gene will be inserted. DNA ligase is used to "glue" twosections of DNA together.

However, the isolated gene cannot be directly inserted into the target DNA. A cloning vector must be used. In the Cohen andBoyer experiment, aplasmidwas used. A plasmid is a circular section of DNA which can be inserted into acell. The cell thentakes on the characteristics of this DNA. This technique is easy, however it does not work on larger animals. The other cloningvector is avirus. The isolated gene is inserted into the DNA of a virus and "glued" using DNA ligase. The virus then injectsthe gene into the cells main DNA. The cell then begins producing the desired protein. Cloning is looked upon as a much easier,cost effective, and reliable way of mass producing genetically altered larger organisms.

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