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Recombinant DNA Technology -

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New idea for generating recombinant DNA conceived

With the use of recombinant DNA human insulin can now be synthesized in the laboratory using eukaryotic or prokaryotic translational vectors.

First time potential biohazards of recombinant DNA raised

There are four different nitrogen bases, adenine, thymine, cytosine and guanine. The synthesis of a particular protein such as insulin is determined by the sequence in which these bases are repeated (see fig.

This has been achieved using Recombinant DNA technology.

Herb Boyer (UCSF) studying restriction enzymes, met at a meeting and realized that they could use restriction enzymes to cut both plasmid DNA as well as DNA containing a gene of interest, and combine the DNAs so that the "sticky ends" of each DNA could be joined, or "spliced", to make a recombinant DNA (ie bacteria ­ human).

Three types of RNA are formed during transcription: (mRNA), (rRNA), and (tRNA). These three types of RNA differ in function, size, and percentage of the total cell RNA (). mRNA makes up only a small percent of the total amount of RNA within the cell, primarily because each molecule of mRNA exists for a relatively short time; it is continuously being degraded and resynthesized. The molecular dimensions of the mRNA molecule vary according to the amount of genetic information a given molecule contains. After transcription, which takes place in the nucleus, the mRNA passes into the cytoplasm, carrying the genetic message from DNA to the ribosomes, the sites of protein synthesis. In , we shall see how mRNA directly determines the sequence of amino acids during protein synthesis.

First paper published on generating recombinant DNA

A recombinant plasmid containing a full length hGH cDNA (which fails to express) is cleaved with restriction enzymes that release a fragment containing the complete hGH coding sequence after codon 24.

are cellular substructures where proteins are synthesized. They contain about 65% rRNA and 35% protein, held together by numerous noncovalent interactions, such as hydrogen bonding, in an overall structure consisting of two globular particles of unequal size.

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  • Recombinant DNA produced in bacteria

    First time possible biohazards of recombinant DNA technology publicly discussed

  • First transgenic mice made with recombinant DNA announced

    First easy-to-use technique published for constucting recombinant DNA. J. Mertz, R. Davis, 69/11, pp. 2270-74.

  • First recombinant DNA based drug approved

    First concerns about potential biohazards of recombinant DNA published

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- Recombinant DNA, Scientific American Books, New York, 1992.

coli bacterial cell produce an insulin chemically identical to its naturally produced counterpart
more reliable and sustainable
Insulin
B-galactosidase
enzyme that controls the transcription of the genes

the insulin gene needs to be tied to this enzyme to make the bacteria produce insulin
Restriction enzymes

Naturally produced by bacteria, act like biological scalpels,
only recognizing particular stretches of nucleotides, such as the one that codes for insulin.

Introduction
Genetic engineer's toolbox

Recombinant DNA Technology in the Synthesis of Human Insulin
variations in diabetic person's blood glucose levels, compared with healthy individuals
Insulin is a small, simple protein.

After Emery AEH, Recombinant DNA technology.

The nucleus contains all the necessary enzymes, proteins, and nucleotides required for this synthesis. A short segment of DNA is “unzipped,” so that the two strands in the segment are separated to serve as templates for new DNA. DNA polymerase, an enzyme, recognizes each base in a template strand and matches it to the complementary base in a free nucleotide. The enzyme then catalyzes the formation of an ester bond between the 5′ phosphate group of the nucleotide and the 3′ OH end of the new, growing DNA chain. In this way, each strand of the original DNA molecule is used to produce a duplicate of its former partner (). Whatever information was encoded in the original DNA double helix is now contained in each replicate helix. When the cell divides, each daughter cell gets one of these replicates and thus all of the information that was originally possessed by the parent cell.

After Emery A, 1984, An Introduction to Recombinant DNA technology.

Gene cloning has a diverse range of applications. Where it has proven particularly useful has been in mapping out the human genome, the creation of transgenic animals, and the development of insect-resistant crops. It is also pivotal to genetic tests carried out in forensic science and archaeology as well as in tests for determining hereditary disease and paternity. The technology also forms the backbone of hepatitis and human immunodeficiency virus (HIV) diagnostic tests. Recombinant DNA technology has also proven important to the production of vaccines and protein therapies such as human insulin, interferon and human growth hormone. It is also used to produce clotting factors for treating haemophilia and in the development of gene therapy.

After Emery AEH, An Introduction to Recombinant DNA.

Sixty three nucleotides are required for synthesising the A chain and ninety for the B chain, plus a codon at the end of each chain,signalling the termination of protein synthesis.

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