LABORATORY 5A TRANSFORMING BACTERIA WITH THE pARA-R PLASMID So far in your quest
ID: 189236 • Letter: L
Question
LABORATORY 5A TRANSFORMING BACTERIA WITH THE pARA-R PLASMID So far in your quest to clone a gene you have verified that you have the pARA-R id containing the rfp gene that can make the red fluorescent protein. In this laboratory you will carry out another step of the gene cloning process, which is to transform E. coli bacteria with this plasmid. Using E. coli bacteria that have been pretreated with calcium chloride, you will divide the bacteria into two groups: a control group to which no plasmid is added, and a treatment group to which you add the pARA-R plasmid. After heat-shocking both groups of cells, you will grow them under several different conditions: The control group is grown in the presence of Luria Broth (a medium that . supports bacterial growth). The control group is grown in the presence of Luria Broth and the antibiotic ampicillin The treatment group is grown in the presence of Luria Broth. The treatment group is grown in the presence of Luria Broth and the ( antibiotic ampicillin. . The treatment group is grown in the presence of Luria Broth, ampicillin, and the sugar arabinose. By examining the growth of bacteria under these conditions, you can verify that your procedure worked, and you can identify the bacteria transformed with the pARA-R plasmid. How will you know if you are successful? The bacteria will have a new and highly visible trait: It will now produce red fluorescent protein, which makes the cells red or bright pink! The pARA-R plasmid, which you reviewed in Chapter 2A, is shown again in mseFigure 5A.3Explanation / Answer
Stop and think answers
Ans-3(1) the p+ bacteria are grown in the media having ampicillin and arabinose sugar. Sugar keep them alive working as carbon source. P- bacteria acts as a control group which can be used as comparison with treatment species.
Ans-3(2) heat shock provide unfavorable condition for growth to retain again in normal conditions one might need some time to overcome the unfavorable treatment.
Ans-3(3) 37° is known as room temperature this is the best temperature at which virus and bacteria can grow rapidly so to get a rapid growth we incubate them at 37° Temperature.
Ans-3(4) aseptic conditions are required to avoid any kind of contamination which may affect the results of our test.
Page 4 answers
Ans-2 the red colonies appear only on some plates because only those plate have favorable conditions for the growth of test bacteria. Other plates don't have favorable conditions.
Ans-3 recombinant plasmid have capacity to replicate independent of cell. Multiple plasmid will reduce the risk of no growth. Sometimes a single plasmid is not able to survive and get replicated so multiple plasmid may work as complementary for each other and will favour growth.
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.