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INSULIN (INTRODUCTION) PRODUCTION OF REC.INSULIN HUMAN GROWTH HORMONE(Hgh)
PRODUCTION OF REC.Hgh
RECOMBINANT VACCINE RECOMBINANT HEPATITIS B VACCINE
INSULIN
Hormone Insulin is produced by the Beta cells of
islets of langerhans. Human insulin contains 51 Amino Acids , arranged in 2 polypeptide chains. Chain-A has 21 amino acids while Chain-B has 30 amino acids. Both are held together by Di-sulphide bonds. Occurrence of Diabetes is due to insufficient Insuline.
the pancreases of pigs and cows was used for the treatment of Diabetics Production of Insulin by recombinant DNA Technology started in late 1970s. Basic technique consisted of inserting human insulin gene and the promoter gene of lac operon on to the plasmid of E. coli. In 1986, Eli Lilly company received approval to market Human Insulin under trade name Humulin.
bacteria. The genes for Insulin A chain and B chain are separately inserted to the plasmids of two different E. coli cultures. The lac operon system is used for the expression of both the genes. Presence of lactose in the culture medium induces synthesis of Insulin A and B chains. Insulin chains are isolated and joined together to form a complete human Insulin
Growth hormone is produced by the pituitary gland. Growth hormone regulates overall body growth by increasing the cellular uptake of Amino Acids, and Protein synthesis. Insufficient human growth hormone in childrens leads to pituitary dwarfism.
signal peptide(with 26 amino acid). The signal peptide is removed during secretion to release the active hGH. Signal peptide interrupts hGH production by recombinanant technology. cDNA encoding signal peptide can be cut to solve these problems.
Continue.
Continued. Base sequence in cDNA encoding signal peptide (26 Amino Acids) + neighbouring 24 Amino Acids = Total 50 Amino Acids; cut by R.E.( ECoRI) Now a gene (cDNA) for 24 Amino Acid sequence of hGH is freshly synthesized and ligated to the remaining hGH cDNA. It is attached to a vector and inserted into E.coli bacterium. In this manner hGH can be produced by DNA technology.
RECOMBINANT VACCINES
INTRODUCTON:
Recombinant/purified protein vaccines are based on the concept that humoral immune responses mounted to an infection are often targeted toward specific localized regions on the surface of protein antigens known as epitopes. The recombinant proteins for vaccination are produced by expressing these immunogenic proteins using heterologous expression systems.
The advancement of genomics, proteomics and biotechnology provide us the opportunity to develop safe and more effective vaccines
Vaccination is the most cost-effective method of reducing animal suffering and economic losses due to infectious diseases in animals
SUBUNIT VACCINES
SUBUNIT VACCINES ARE DEFINED AS THOSE CONTAINING ONE OR MORE PURE OR SEMI-PURE ANTIGENS
Cerevisiae).
Gene for HbsAg is inserted which is linked to alcohol dehydrogenase promoter. These plasmids are then transferred and cultured. The cells grown in Tryptophan, free
against Hepatitis B.
& T-Lymphocytes.
2.
Continued..
The plasmid insertion vectors incorporates its
genes into vaccinia virus genome at a place that encodes for the enzyme Thymidine Kinase(TK). This the recombinant viruses have lost their ability to produce TK gene due to which it is easy to select recombined vaccinia viruses that lack TK gene. These viruses are less infectious than the normal viruses. The recombinant vaccinia viruses, release from the cultered animal cells,can be successfully used as vaccines.
REFERENCE
BIOTECHNOLOGY by U. Satyanarayana
IMMUNOLOGY by KUBY www.virology.ws
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