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quiz 4 Micro
MICRO
| Question | Answer |
|---|---|
| what is the purpose of the phenylalanine test | to differentiate Proteus from other Enterobacter |
| phenylalanine slants contain | amino acid--phenylalanine is an amino acid |
| in the phenylalanine test what do microbes with enzyme do | phenylalanine deaminase can remove amino group--producing an acid |
| in the phenylalanine test what happens when an acids reacts with Ferric chloride | green color is produced |
| what were the outcomes of the three tests; the colors/bacteria | 1. Proteus vulgaris- phenylalanine deaminase (+), green 2. uninoculated-control, tan 3. E. coli- phenylalanine deaminase (-), tan |
| what is the purpose of the urea hydrolysis test | is used to differentiate organisms based on their ability to hydrolyze urea-Urinary tract pathogens from the genus Proteus can be distinguished from other enterics by their rapid urease activity |
| in the urea hydrolysis test most enterics do what | hydrolyze urea--just not rapidly |
| what does the enzyme urease do | breaks urea down into ammonia and C02--release of ammonia raises pH |
| urea broth contains | urea, pH indicator, bufferes |
| what is the pH indicator in the urea hydrolysis test | phenol red dye, Yellow-Red/Orange-Pink |
| what is the buffer in the urea hydrolysis test | inhibit alkalization of medium by all microbes except rapid urease (+) microbes like Proteus vulgaris |
| what were the test results of the urea hydrolysis test | 1. Proteus vulgaris-rapid urease (+), pink 2. unioculated- control, orange 2. E. coli-slow urease (+), orange |
| what is the purpose of the transformation test | process by which "competent" bacterial cells pick up DNA and use the genes to make a protein |
| what are the key components of transformation test | 1. CaCl2 2. E. coli 3. pGlo 4. LB broth |
| CaCl2 transformation | transformation soultion-causes cell wall of E.coli to be permeable to plasmid |
| E.coli transformation | cloning host-makes multiple copies of proteins |
| pGLO transformation | plasmid-vector-inserts genes into E/coli |
| LB broth transformation | nutrients for E.coli |
| what were the genes added to E.coli transformation | 1.green fluorescent protein-causes bioluminescence in jellyfish 2. resistance to antibotic- ampicillin |
| what else was added to the E. coli transformation | 1. replication orgin- allows copies of inserted genes to be replicated in every generation 2. arabinose promoter- attachment site for RNA polymerase |
| how does the transformation test work | 1. in presence of arabinose, the regulatory gene turns the operon on 2. instead of producing enzymes to break down arabinose, the operon codes for green fluorescent protein |
| what does the media contain in the transformation test | 1. amp=ampicillin-antibotic inhibits growth of E.coli 2. ara=arabinose-monosaccharide that turns on the operon |
| +DNA LB/amp-growth-glo in dark | Yes-E.coli received he genes to resist amplicillin. No-no arabinose(inducer) to turn on the operon |
| +DNA LB/amp/ara-growth-glo in dark | Yes-E.coli received he genes to resist amplicillin. Yes- media has arabinose-turns on the operon-E.coli starts making green fluorescent protein (GFP) |
| -DNA LB/amp-growth-glo in dark | No- media contains ampillican, which kills E.coli. No- E.coli did not get genes to glo in the dark |
| -DNA LB-growth-glo in dark | Yes- no amplcillin in the media. No- E.coli did not get genes to glo in the dark |
| what was the purpose of transferring the +DNA and -DNA from ice to hot water to ice | heat shock- rapid change in temp make E.coli cell wall permeable to plasmid DNA |
| why were the vials incubated for 10 mins in LB broth rather than transferring their contents directly to the plates | 10 mins in LB broth gives transferred cells time to grow- immediately begins producing ampicillin resistance gene--if placed directly on plates bacteria would be killed |
| what information is provided by the LB/-DNA plate | control-verfies that E.coli can grow on LB media without ampicillin |
| which plates exhibit transformation | +DNA LM/amp, +DNA LB/amp/ara |
| what information is provided by the LB/amp/-DNA plate | control-verifies that E.coli cannot grow on media containing ampicilliin |
| why does the LM/amp/ara +DNA plate fluoresce when the LB/amp/+ DNA plate does not | GFP-is only expressed in the presence of arabinose(inducer)-turns on operon |
Created by:
Khuller