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Genetic engineering

• A gene product is any biochemical material derived from the extraction and/or manipulation of a particular gene from the genome of an organism. • Gene products are produced by genetic engineering • The branch of genetic engineering used to make gene products is known as Recombinant DNA Technology. This includes several techniques:

.Recombinant DNA Technology Techniques: Restriction analysis. involving the use of restriction enzymes DNA sequencing (where the sequence of nucleotides is known and the gene can be synthesized in the laboratory) Nucleic acid hybridization (used to locate the target gene on its chromosome) DNA cloning (the gene is inserted into the genome of a foreign organism (host) and is replicated as the cells in the host divide).

Molecular Cloning (Process by which gene products are made) PCR product cloning vector ligation transformation expression E. coli protein 3 .

Steps in making gene products by Recombinant DNA Technology: • Identify the gene • Locate the gene on its DNA strand using an appropriate Polymerase Chain Reaction or Nucleic Acid Hybridization technique. • Isolate the gene (this process involves several steps including the use of restriction enzymes). • Ligate the gene with a cloning vector • Transform the ligated gene into a host and allow it to replicate. • Extract the gene and insert it into the desired genome .

Examples of Gene Products Insulin The insulin gene is found in the human genome. It is needed for carbohydrate metabolism and insulin deficiency results in diabetes. .  The objective of Genetic engineering in this case is to clone the insulin gene from the human genome.

Fig.2 The human insulin gene . 1.

insulin from the pig and cattle genomes were administered. coli genome after translation and is replicated as the bacterium’s cells divide. yeast played this role. coli bacterium.In earlier years. The mass produced insulin can then be extracted and given to the diabetic patient. The host in which the replication of extracted insulin occurs is the E. They however were not as effective as the modern genetically engineered variety. .  Before genetic insulin production. The insulin gene becomes integrated into the E.

It governs the synthesis of the human albumin protein in the liver(The most abundant protein in blood plasma and aids in the transport of thyroid hormones. fatty acids.Human Albumin The human albumin gene. protein-losing enteropathy cause low albumin (hypoalbuminemia)in humans. . This can be treated by the administering of genetically engineered Human Albumin Gene. Liver disease. other complex molecules. like the human insulin gene exists in the human genome. nephrotic syndrome.

This however is not the only means of treatment. It is then transformed into the host’s E. the genetically engineered human albumin gene is extracted from the host and can be administered to a patient suffering from hypoalbuminemia.coli’s genome in which it replicates normally. . After sufficient replication.The gene when extracted from the human genome is ligated unto the cloning vector. bacteriophage (lambda).

Persons deficient of factor VIII suffer from a form of haemophillia know as haemophillia A.Factor (VIII) Eight  Factor VIII is produced naturally in humans by the factor VIII gene. . It is known as an anti-haemophillic factor as it aids in the blood clotting process. as their factor VIII gene is defective in some way.  The factor VIII gene after extraction from a human donor is ligated into the bacteriophage  FVIII23.

Factor VIII is produced in the host as its cells divide and when sufficient amount is produced it can be extracted. The ligated gene in then transformed into hamster kidney cells which act as the host. Factor VIII is afterwards purified by heat treatment before being given to a haemophilliac patient during therapeutic treatment. . during production.

Elastase is an enzyme produced by some white blood cells which destroys elastic fibres in the lungs as a part of the normal turnover of elastic tissue. When the gene that makes AAT is isolated from a human then is placed into hemagglutinin (HA)-tagged vector pcDNA3-HA. This vector is then transformed into eukaryotic cells.Alpha-1-antitrypsin (AAT) • Alpha 1-Antitrypsin or α1-antitrypsin (A1AT) is a protease inhibitor belonging to the serpin superfamily. It is a naturally occurring protein which is found in human blood. AAT is however made in the liver and can be extracted from the blood . more specifically the mammary glands of a sheep to express the gene. • • • • . A mutant form of the gene that codes for it causes a genetic disease leading to emphysema as a result of uninhibited elastase activity. however more people need it than can be supplied by normal means.

• But it is not confirmed that the gene will be expressed into the transgenic offspring • For the gene to be expressed the promoter. the piece of DNA next to the gene must be turned on • By cloning the promoter beta-lactoglobulin and attaching it to the human DNA we can ensure that only mammary glands express that gene .

AAT rendering .

tPA is used in clinical medicine to treat only embolic or thrombotic stroke.Tissue Plasminogen Activator (Tpa) • Tissue plasminogen activator (abbreviated TPA or PLAT) is a protein involved in the breakdown of blood clots. • tPA may be manufactured using recombinant biotechnology techniques. the major enzyme responsible for clot breakdown.4. It is a serine protease (EC 3. Use is contraindicated in hemorrhagic stroke and head trauma.68) found on endothelial cells.21. it catalyzes the conversion of plasminogen to plasmin. tPA created this way may be referred to as recombinant tissue plasminogen activator (rtPA). As an enzyme. Because it works on the clotting system. . the cells that line the blood vessels.

tPA rendering of the actual protein .

• The retroviral vectors derived from murine leukemia virus (MuLV) were used to transfer human tPA cDNA • The host cells are Eurkaryotic and have been tranfered to transgenic mice to extract the tPA. .