COSMIDS:

Cosmid is a hybrid DNA formed by the joining of a plasmid and lambda phage DNA carrying a Cos site (cohesive). In brief cosmid is a plasmid carrying the Cos Site of a L-phage DNA. Cosmid is not a naturally found in living cells. It is constructed vector. E.g. Col E1 cosmid is a typical cosmid used in genetic engineering. CONSTRUCTION OF COSMIDS: Col E1 cosmid is a constructed from Col E1 plasmid and L phage Dna. The plasmid is cut with restriction endonuclease, which removes a portion of DNA form the plasmid. Te same restriction enzyme is used to cut the L phage DNA to get a DNA fragments containing Cos site. These two DNA fragments are mixed together in the presence of enzymes DNA ligase, which linked together the two DNA fragments end to end. The resulting recombinant plasmid is called Col E1 cosmid. CHARACTERISTIC FEATURES: 1. It is a circular double stranded DNA. 2. It contains a complementary single strand regions, these regions are abbreviated as Cos site. 3. The cosmids DNA dose not participate in the multiplication of phage particles. Advantages: 1. The cosmids transfer somewhat larger foreign gene into the bacterial cell. 2. The cosmids pick up even long size genes. Hence it is used for the entire genome of organisms. 3. Loading capacity of csosmid is usually 40,000-45,000 base pairs.

Disadvantages: 1. Each cosmid requires 2 Cos sites for the successful packing of recombinant cosmids. 2. The packing fails when the distance of separation exceeds 54000 base pairs or when it is less then 38,000 base pairs. Lambda phage of Lambdiod phage: 1. Lambda is a temperate bacteriophage with a genome size of about 48.5 kilo base pairs. It has a specific susceptibility for F coli. Its entire DNA sequence is known. The term temperate means it enjoys one of two life cycles. Lytic cycle replicate many times,

produces more phage. It can also take up lysogenic growth meaning that it migrates its DNA into bacterial cell.(further line is miss printed). 2. Lambda genome exists as a linear double stranded molecule with a single stranded complementary ends. 3. The points at which the segments of bacteriophage lambda are cut are called as Cos (Cohesive sites). 4. The Cos sites are the only part of lambda that is included in the plasmid of a cosmid vector. No viral gene is included in the vector when the plasmid is introduced to E-coli.

GENE SILENCING
Interruption or suppression of the expression of a gene at transcriptional or translational levels is called as gene silencing. As the most of the emphasis of genetic engineering efforts had been traditionally to increase the expression of gene. Especially the useful ones gene regulation and gene silencing has helped in this regards. 1. Gene silencing can take both at the transcriptional as well as translational levels. 2. Transcriptional gene silencing takes place due to modification of DNA mainly by methylation of histones by acetylation. 3. These changes remarkably affect the transcription of those stretches of DNA, usually by reducing the transcription initiation at the promoter. 4. Post transcriptional gene silencing takes place at the RNA level. Here specific transcripts of genes are degraded by the special complex called Dicer. 5. Dicer consists of Ribonuclease. 6. Dicer molecules quickly degrade specific transcripts there by reducing their levels and bringing about the silencing even through the transcription continues normally. 7. Because post transcriptional gene silencing is involved in targeting individual transcripts thus it can be used efficiently to silence the required genes and to understand their functions in the body. 8. This technique can also be used for silencing disease causing gene. E.g. cancer. Diagram:

APPLICATIONS OF GENETIC ENGINEERING

1. Many of the genetic diseases like haemophilia, sickle cell anemia etc, can be treated using gene therapy. 2. Its due to genetic engineering that industrial production of vaccines like insulin, interferons is possible which is a real blessing for mankind. 3. By using a recombinant technology, scientists have inserted genes into bacteria for the production of human growth hormone. This hormone is used to treat dwarfism. In 1986 the hormone becomes commercially available as Protropin. 4. Urokinase a clot dissolving enzyme is produced from genetically engineered bacteria which is a savior for angina patients. 5. Since microbial cells have much higher metabolic rates, genes of desired enzymes (of commercial values) could be introduced into plasmid of bacteria. E.g. yeasts are being engineering to yield enzymes for cheese industry. 6. Attempts are being made to engineering plants with bacterial genes that trap Nitrogen (N2) and convert it to a form plants. This would eliminate the necessity of nitrogen fertilizers. 7. By using gene expression silencing and tracking researchers have revolutionized in the field of recombinant technology. Now they are able to find out more genes and are able to remove single deleterious allele. 8. Genetic engineering is being used to create genetically modified food and genetically modified crops. These crops are resistant to herbs, sherbs, and fungal viruses. 9. In materials science, genetically modified virus has been used to construct a more environment friendly lithium ion battery.