BIO208 Bacterial Genetics Worksheet 1 1. Fill in: Transformation Transduction Conjugation a. _______________a. naked DNA is involved b. _______________b. faulty head stuffing may occur c. _______________c. involves exconjugants d. _______________d. adsorption and injection of genetic material e. _______________e. competent cells required f. _______________f. an F plasmid encodes attachment proteins g. _______________g. used to construct the E. coli minute map h. _______________h. a pilus is produced i. _______________i. lytic cycle entered in response to stress j. _______________j. may be specialized or general k. _______________k. requires cell to cell contact l. _______________l. mediated by a virus m. _______________m. known as parasexual mating n. _______________n. requires competent cells o. _______________o. involves a phage 2. Fill in - you may use a term more than once, or not at all homologous recombination extrachromosomally (episomally) F+ cell F Factor (plasmid) Hfr strain F minus (F-) generalized phage packaging lysogeny adenovirus adsorption a. the reciprocal exchange of DNA between two DNA fragments of identical (or at least close to identical) sequences is called __________________________________ b. A plasmid can be maintained _______________________ or integrate into bacterial chromosome. c. In ______________ transduction, all bacterial DNA has equal probability of being packaged into progeny phage d. In ________________, the phage genome integrates into bacterial genome creating a prophage. e. The ______________ contains ~100,000 base pairs and contains 19 genes that encode for proteins involved in pilus synthesis and other functions f. One method of gene therapy uses replication deficient _____________ to deliver a gene to body cells g. The F factor occasionally integrates randomly into the E. coli chromosome creating a _______ h. In normal conjugation, recipient (female) cells remain __________ i. After a phage engages in _____________, it will inject genetic material into bacterium j. Lambda bacteriophage assembles new virus during the process of ________________ k. The creation of a knockout mouse uses the process of ________________ to replace DNA with an engineered construct 3. What is the difference between: a. A bacterial colony and a lawn b. The log phase and the stationary phase of bacterial growth c. Minimal media and complete media d. A conditional mutant and a nutritional mutant e. Binary fission and parasexual mating f. A prototroph and an auxotroph g. A plasmid and a chromosome h. Transduction and transformationBIO208 Bacterial Genetics Worksheet 2 i. A phage and a bacterium j. An F+, an Hfr, and an F- cell k. A donor and recipient cell l. A U tube and a blender m. Size of F factor and size of bacterial genome n. Lytic and lysogenic cycle o. A virulent and a temperate phage p. Prophage and phage q. Phage adsorption and phage packaging r. A cfu and a plaque 4. What is the difference between a. operon inducer and repressor b. betagalactosidase and Z structural gene c. promoter and operator d. O and Oc e. I and I- f. lactose and glucose g. negative and positive operon regulation h. polycistronic and monocistronic (as in humans) mRNA i. constitutively expressed and regulated genes 5. Operons are found in (circle one) a. Prokaryotes b. Eukaryotes 6. Fill in using the following choices lactose repressor protein operator lac Z promoter beta galactosidase a. _____________________________ DNA that binds repressor to prevent transcription b. _____________________________ DNA that binds RNA polymerase c. _________________________ binds repressor to allow transcription to occur d. _____________________________ a structural gene of the operon e. _____________________________ is bound by lactose f. _____________________________ a trans acting protein that represses the operon g. _____________________________ encoded by the I gene h. __________undergoes a conformational (shape) change which prevents binding to the operator i. _____________________________ gene that is actively transcribed when derepressed j. _____________________________ enzyme that cleaves lactose into glucose and galactose k. _____________________________ 3 DNA elements of the lac operon l. _____________________________ 2 proteins associated with the lac operon m. _____________________________ 1 sugar associated with the lac operon 7. 7A normal bacterial cell is grown in lactose (sugar) in the absence of glucose. The lac operon is: a. off (repressed) b. on (induced) 8. A bacterial cell has a lactose operon but the promoter is defective. All else is normal. The bacterium is transformed with a plasmid that contains a wild type (normal) promoter, amp resistance gene, and origin of replication. Can the cell utilize lactose? (i.e. is the operon inducible?) Why or why not?BIO208 Bacterial Genetics Worksheet 3 9. 9. Match the plasmid feature to the gene. You will use some answers more than once. _____ allows cell to deactivate ampicillin A. ara C _____ allows plasmid to replicate autonomously B. bla gene _____ also called ampR C. ori _____ allows cells to manufacture GFP D. GFP gene _____ regulates the expression of GFP gene E. multiple cloning site _____ enables genes to be ligated into plasmid _____ known as a reporter gene _____ recognized by a variety of restriction enzymes _____ a foreign gene not normally found in bacteria _____ enables jellyfish and other marine organisms to glow 10. 36 bacterial colonies were observed on a nutrient agar plate. The original sample was 0.1 ml withdrawn from the culture and diluted 1:1000 prior to plating. Determine the titer (cfu/ml). 11. Why is it not useful to use Salmonella bacteria in phase C? 12. Growth curve of Salmonella