GMS6038-2010-final-key

GMS6038-2010-final-key - Bacterial Genetics GMS6038 Final...

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Bacterial Genetics GMS6038 Final Exam Fall 2010 Write your station code here. ___________________________ You have 2 1/2 hours to complete the exam. Please note - this is a closed book, closed note exam. All backpacks and notebooks must be in the cubbies against the wall. All cell phones and personal communication devices must be off and put away. You may use the rest room one person at a time. Any cheating will result in a 0 for the exam and failure of the course. This exam is in the form of a Word document. Simply type your answers below the questions. You may write on the paper form of the exam, but you will have to turn it in at the end of the exam. The exam center personnel will instruct you on mechanistic of the computer system.
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Shown below is pGULIG-12. Note the following genetic elements: At the top of the map is the origin of replication of the ColE1 plasmid. Note that the rna-II is expressed from the trp promoter. Note that the trp promoter is not followed by any trp protein coding sequences. Moving clockwise: From 1:00-3:00 are the genes for the coding sequences for the two components of a typical two partner secretion system. The β barrel protein and passenger protein genes are indicated. Both of these genes are expressed by the promoter for the araBAD genes. From 4:00-5:00 is the tet gene expressed by the λ pR promoter. From 6:00-8:00 is the pir gene expressed from the dnaK promoter. From 8:00-10:00 is the λ CI gene expressed from the lac promoter. There are no other promoters on this plasmid other than those indicated on the map. Assume that all genes have appropriate translation start and stop codons. The short bars crossing the circle represent typical factor-independent terminators that will prevent transcriptional read through between these different elements. The host E. coli strain lacks the F plasmid and λ phage . Any E. coli genes that normally regulate the promoters on this plasmid are not encoded on this plasmid unless indicated.
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NOTE: These questions involve 2 or 3 sentence answers - not paragraphs. The number of points is the maximum number of facts being looked for, and in some cases one fact is worth 2 points. Keep to what is being asked. Adding extra material hoping to include the right answer somewhere in the middle could count against you if your answer reveals a lack of understanding or includes incorrect information, even if unrelated to the original question. The following questions are based on pGulig-12. 1. (6 points) How would you change the copy number of pGULIG-12 by altering growth conditions? Explain for both increasing and decreasing copy number. Be sure to detail how this works at the molecular level. There is only one origin of replication on the plasmid, the ori of the ColE1 plasmid. It is an RNA-based origin, so the promoter of the initiating RNA, RNA-2, determines the replication and copy number. The promoter is the trp promoter without any coding sequences, i.e., trpL, the leader protein that encodes the attenuation system. So this promoter is regulated strictly
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This note was uploaded on 12/13/2011 for the course GMS 6038 taught by Professor Gulig during the Fall '11 term at University of Florida.

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GMS6038-2010-final-key - Bacterial Genetics GMS6038 Final...

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