Black Chapter 8-Gene transfer and genetic Engineerings THE TYPES AND SIGNIFICANCE OF GENE TRANSFER Gene transfers

refers the movement of genetic information between organisms. Vertical gene transfer passes genes from parent to offspring. Lateral gene transfer passes genes to other cells in the same generation. It occurs in bacteria by transformation, transduction, and conjugation. Gene transfer is significant because iy increases genetic diversity within a population, thereby increasing the likelihood that some members of the population will survive environmental changes. TRANSFORMATION The discovery of Transformation Bacterial transformation was discovered in 1928 by Griffith, who showed that a mixed culture of live rough and heat-killed smooth pneumococci could produce live smooth pneumococci capable of killing mice. Avery later showed that a capsular polysaccharide was responsible for virulence and that DNA was the substance responsible for transformation Watson and Crick determined the structure of DNA, which led to studies showing that a cells genetic information is encoded in its nucleic acids. The Mechanism of Transformation Transformation involves the release of naked DNA fragments and their uptake by other cells at a certain stage in their growth cycle; (1) Uptake of DNA requires a protein called competence factor to make recipient cells ready to bind DNA. (2) Endonucleases cut double-stranded DNA uinto units; the strands separate, and only one strand is transferred. (3) Ultimately, donor DNA is spliced into recipient DNA. Leftover recipient DNA is broken down, so a cells total DNA remains constant. The Significance of Transformation Transformation is significant because (1) it contributes to genetic diversity (2) it can be used to introduce DNA into an organism, observe its effects, and study gene locations; (3) it can be used to create recombinant DNA. TRANSDUCTION The Discovery of Transduction In transduction, genetic material is carried by bacteriophage (phage). The Mechanism of Transduction Phages can be virulent of temperate. (1). Virulent phages destroy a host cells DNA, direct synthesis of phage particles, and cause lysis of the host cells in the lytic cycle.

(2). Temperate phages can replicate themselves as a prophage-part of a bacterial chromosomes-or eventually produce new phage particles and lyse the host cell. Persistence of the phage in the cell without the destruction of the host cell is called lysogeny. Prophage can be incorporated into the bacterial chromosome, or it can exist as a plasmid, a piece of extrachromosomal DNA. Cells that containsa prophage are called lysogenic cells because they have the potential to enter the lytic cycle. Transduction can be specialized or generalized. (1) In specialized transduction, the phage is incorporated into the chromosome and can transfer only genes adjacent to the phage. (2) In generalized transduction, the phage exists as a plasmid and can transfer any DNA fragments attached to it. The Significance of Transduction Transduction is significant because it transfers genetic material and demonstrates a close evolutionary relationship between prophage and host cell DNA. Also, its persistence in a cell suggests a mechanism for the viral origins of cancer, and it provides a possible mechanism for studying gene linkage. CONJUGATION The Discovery of Conjugation In conjugation large quantities of DNA are transferred from one organism to another during contact between donor and recipient cells. Conjugation was discovered by Lederberg in 1946 when he observed that mixing strains of E. coli with different metabolic deficiencies allowed the cells to overcome deficiencies. Plasmids are extrachromosomal DNA molecules. The Mechanisms of Conjugation Three mechanisms of conjugation have been observed: (1) In the transfer of F plasmids, a piece of extrachromosomal DNA (a plasmid) is transferred. (2). In high-frequencccy recombinations , parts of F plasmids that have been incorporated into the chromosome (the initiating segment) are transferred along with adjacent bacterial genes. (3) An F plasmid incorporated into the chromosome and subsequently separated becomes an F plasmid and transfers chromosomal genes attached to it. The Significance of Conjugation The significance of conjugation is that it increases genetic diversity, it may represent an evolutionary stage between asexual and sexual reproduction, and it provides a means of mapping genes in bacterial chromosomes. Promiscuous - those can transfer to other species. Gene Transfer Mechanisms Compared

Genetic transfer mechanisms differ in the quantity of DNA transferred. PLASMIDS Characteristics of Plasmids Plasmids are circular, self-replicating, double-stranded extrachromossomal DNA that carry information that is usually not essential for cell growth. Resistance Plasmids Resistance (R) plasmids carry genetic information that confers resistance to various antibiotics to certain heavy metals. They generally consist of a resistance transfer factor (RTF) and one or more Resistance (R) genes. Transposons R genes that move from one plasmid to another in a cell or become inserted in the chromosome are part of a transpons because they transpose, or change, their locations. Bacteriocinogens Bacteriocinogens are plasmids that produce bacteriocins, which are proteins that inhibit growth of other strains of the same species or closely related species. GENETIC ENGINEERING Genetic Engineering is the manipulation of genetic material to alter the characteristics of an organism. Genetic Fusion Genetic fusion allows transposition of genes from one location on a chromosome to another, sometimes deleting a portion, thereby causing the jointing of genes from two different operons. Protoplast Fusion Protoplast Fusion combines protoplasts (organisms without cell walls) and allows mixing of genetic information. Gene Amplication Gene amplication involves the addition of plasmids to microorganisms to increase yield of useful substance. Recombinant DNA Technology Recombinant DNA is DNA produced when genes from one kind of organism are introduced into the genome of a different kind of organism. The resulting organism is a transgenic, or recombinant organism. Recombinant DNA has proven especially useful in medicine, industry, and agriculture. 1).The donor segments are incorporated into a vector (plasmid) 2) It can be introduced into cells by heating in a solution of calcium chloride or by electroporation- use electrical pulses to produce pores in the cell membrane. 3). Restriction endonuclease (restriction enzymes) cut specific DNA sequences create restriction fragment.

4). Restriction Fragment length polymorphisms (RFLPs) DNA sequences can be slightly different between members of the same species. Medical Applications of Recombinant DNA 1). To make bacterial cells produce human proteins, a human DNA (synthesize the protein) is inserted into vector. Interferon (treat viral diseases and cancer) and insulin, human growth hormone, vaccines can be produced. 2). Diagnosis of genetic defects in a fetus by studying enzymes in fetal cells from amniotic fluid. By using recombinant DNA with a known nucleotide sequence to find errors in the nucleotide sequence in fetal DNA segments. 3). Gene therapy : inserting the gene in a defective gamete (egg or sperm) might prevent offspring from inheriting a genetic disease, like in severe combined immunodeficiency disease. 4). Make a blood substitute Inject copies of human hemoglobin genes into 1-day-old pig embryos that had been removed from their mothers uteri. The embryos were then implanted into a second pigs uterus to grow until delivery. Two days after birth, the piglets were tested to see whether they produced human hemoglobin with pig hemoglobin (transgenic, 0.5% succeed).The transgenic pigs produced blood containing 50% human hemoglobin. The pigs are bled and red blood cells are ruptured and purified. It has storage life of months instead weeks. Because naked hemoglobin does not have any antigens like red blood cells, it can be transfused without blood typing. It can ensure safety from human pathogens, and use as immediate sources of oxygen during disaster. Industrial Application of Recombinant DNA Fermentation processes used in making wine, antibiotics might be greatly improved by the use of recombinant DNA. Agricultural Application of recombinant DNA Some bacteria are being engineered to control insects that destroy crops. Pseudomonas fluorescens (colonizes the roots of corn) has been induced to carry genetic information inserted into it from Bacillus thuringiensis, allowing P.fluorescens to synthesize a protein that kills insects. Toxins made by B.thuringiensis has been extracted and used as insecticide. Pseudomonads could replace the use of chemicals insecticides to control black cutworm and other insect larvae that damage crops. Hybridomas Fusion of a myeloma (bone marrow cancer) cell with an antibody-producing white blood cells. It can produce monoclonal antibodies. Weighing the Risks and Benefits of Recombinant DNA When recombinant DNA techniques were first developed, scientist were concerned that virulent pathogens might be created, and they developed containment procedures. As research proceeded and no illness caused by recombinants were observed, most scientists came to believe that the benefits of recombinant DNA techniques outweigh the risks.