Microbiology Exam 4 Study Guide
Microbiology Exam 4 Study Guide BIO 2200 (Microbiology, Dr. Thomas Roberts)
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BIO 2200 (Microbiology, Dr. Thomas Roberts)
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This 3 page Study Guide was uploaded by Taylor Notetaker on Monday April 11, 2016. The Study Guide belongs to BIO 2200 (Microbiology, Dr. Thomas Roberts) at Wayne State University taught by Dr. Thomas Roberts in Winter 2016. Since its upload, it has received 44 views. For similar materials see Microbiology in Biological Sciences at Wayne State University.
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Date Created: 04/11/16
Exam 4 Study Guide What are restriction enzymes? - recognize specific DNA sequences & cut DNA at those sites - protect prokaryotes from hostile foreign DNA What are modification enzymes? - protect cell’s DNA for restriction enzymes - chemically modify nucleotides in restriction recognition sequence What are the 2 categories of antimicrobial drugs? - synthetic antimicrobial drugs & antibodies What are the fluorescent methods? - direct method: the antibody targeted against the surface antigen is covalently linked to the fluorescent dye - indirect method: the presence of a nonfluorescent antibody on the surface of a cell is detected by use of a fluorescent antibody directed against the nonfluorescent antibody True or False You must have Gel Electrophoresis if your have PCR. FALSE - You must have PCR in order to have Gel Electrophoresis but you don’t have to have Gel Electrophoresis if you have PCR because PCR is amplification of the DNA and you can’t perform Gel electrophoresis without a ample amount of DNA. True or False Nucleic acids migrate through Gel toward the positive electrode. TRUE True or False PCR (also know as DNA amplification) is a process of replication in a test tube. TRUE True or False Plasmids are synthetic vectors & have useful properties as cloning vectors. FALSE - Plasmids are NATURAL vectors not synthetic vectors. True or False pUC19 is a common cloning vector. TRUE True or False Insulin was the 1 synthetic protein made commercially by genetic engineering. FALSE st st - Insulin was the 1 HUMAN protein not the 1 synthetic protein. Southern blot: a hybridization procedure where DNA is in the gel and the probe is DNA or RNA Northern blot: RNA is in the gel and the probe is DNA or RNA Western blot (immunoblot): methods detect antibodies to specific antigens or the antigens themselves Steps in PCR: add DNA add primer add DNA polymerase (Taq polymerase or Pfu polymerase) heat & cool (2x) Variations of PCR Reverse transcription (RT PCR) – can make DNA from an RNA template Quantitative PCR (q PCR) – uses fluorescent probe to monitor the amplification process - same as RT PCR but it quantifies how much you started with 3 Steps of Gene Cloning 1. isolation & fragmentation of source DNA 2. insertion of DNA fragments into cloning vector 3. introduction of cloned DNA into host organism Site directed mutagenesis: can be used to assess the activity of specific amino acids in a protein Cassette mutagenesis: replacing a fragment you don’t want with new synthetic one - form of gene disruption (occurs when cassettes are inserted into the middle of the gene) o causes knockout mutations Blue colonies DON’T have vectors with foreign DNA inserted. White colonies HAVE foreign DNA inserted. T7 RNA polymerase recognizes only T7 promoters. - transcribes only cloned genes - shuts down host transcription Cloning with lambda 1. isolate vector DNA from phage particles, & cut it with the appropriate enzyme 2. connect the lambda fragments to foreign DNA using DNA ligase. 3. Package the DNA by adding cell extracts containing the head & tail proteins 4. Infect E. Coli cells & isolate phage clones by picking plaques. 5. Check the recombinant phage for the presence of foreign DNA. Agrobacterium tumefaciens can be used to introduce DNA into plants. - contains the T1 plasmids, which is responsible for virulence - T1 plasmid contains genes that mobilize DNA for transfer to the plant - Segment of the T1 plasmid that is transferred to the plant is T- DNA Recombinant vaccines - vector vaccine: vaccines made by inserting genes from a pathogenic virus into a relatively harmless carrier virus (ex: vaccinia virus) - subunit vaccine: contain only a specific protein or proteins from a pathogenic organism (ex: coat protein of a virus) Polyvalent vaccines: a single vaccine that immunizes against 2 different diseases Differential media incorporate biochemical tests to measure the presence or absence of enzymes involved in catabolism of specific substrates. Two common fluorescent dyes are rhodamine B (red) & fluorescent isothiocyanate (yellow-green). 4 types of EIA methodologies 1. direct EIA: detection of antigen 2. indirect EIA: detection of antibody 3. sandwich EIA: detection of antibody 4. combination EIA: combination of direct EIA & sandwich EIA
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