Why do we use Ampicillin only for transformation of BL21(DE3) cells and Amplicillin plus chloramphenicol for...
Why do we use Ampicillin only for transformation of BL21(DE3) cells and Amplicillin plus chloramphenicol for BL21(DE3)pLysS cells?
Homework Answers
Answer to why do we use Ampicillin
In transformation process we use antibiotic (here it is Ampicillin) to select or grow only transformed cells. In a similar fashion here Ampicillin being used in case of BL21(DE3) cells and BL21(DE3)pLysS cells.
Now,
Answer to Why do we use only Ampicillin in BL21(DE3) cells but both ampicillin and chloramphenicol in BL21(DE3)pLysS cells
The difference between these two type of cells is BL21(DE3)pLysS contains one extra plasmid known as pLysS plasmid.
Chloramphenicol is added only to maintain this plasmid. This plasmid is very helpful in tighter control of protein expression as well as in toxic recombinant protein expression and also it reduces basal protein expression. And if chloramphenicol is not added during transformation process then pLysS plasmid will be lost and the cells will be same as BL21(DE3) cells.
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Why do we use Ampicillin only for transformation of BL21(DE3)
cells and Amplicillin plus chloramphenicol for...
The BL21(DE3) expression cells carry a λDE3 integration containing the T7-RNA polymerase gene. A) Why is...
The BL21(DE3) expression cells carry a λDE3 integration containing the T7-RNA polymerase gene. A) Why is the T7-RNA polymerase protein so specific with regard to the target genes it transcribes? B) Is this enzyme predicted to transcribe any E. coli genes? Why or why not?
Answer the following questions: a) What is special about the E coli BL21(DE3)pLysS cells used for...
Answer the following questions: a) What is special about the E coli BL21(DE3)pLysS cells used for gene expression? b) In what structural form is the expressed gdhA gene synthesized in the E coli host cells? c) What physical attributes of SDS-PAGE gels allow the separation and molecular weight determination of polypeptides? d) Given the size of the gdhA gene of ~1362 bp, what is the theoretical size of the expressed protein? e) Does your answer from (d) agree with the...
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where is the bacteria usually found? Why do we use it in transformation experiments?
where is the bacteria usually found? Why do we use it in transformation experiments?
3. A) Why is it necessary to include ampicillin in the medium with the (-) plasmid...
3. A) Why is it necessary to include ampicillin in the medium with the (-) plasmid treatment? B) of the (+) plasmid treatment? the two genes of interest on the plasmid were only expressed on the plate with ampicillin. Why? 15. What causes the "halo effect", the white satellite colonies around the green fluorescent colonies, in the (+) plasmid TSA + ampicillin plate? (6. Other than transformation, what are two other mechanisms that bacteria use to acquire new genes?
Question 1 Not yet answered Marked out of 1.00 Flag question If we do a "negative...
Question 1 Not yet answered Marked out of 1.00 Flag question If we do a "negative control transformation experiment with no plasmid DNA added, we would expect to see Select one: a. no colonies on our transformation plates b. the same number of colonies as we see with the plasmid DNA added c. more colonies compared to when we add the plasmid DNA QuJLIUI Answer saved Marked out of 1.00 Flag question If we used the wrong antibiotic (kanamycin instead...
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2. Why do broad-spectrum antibiotics such as streptomycin, chloramphenicol, and tetracycline inhibit a wide variety of prokaryotes, but do not harm human cells? a. 3. If an infectious agent is sensitive to several antimicrobial drugs, what other considerations might be taken into consideration to determine which drug should be prescribed to the patient? a 4. Look at a household cleanser or antiseptic that you use in your own home. What claims are made about its effectiveness against microbes (read the...
CRITICAL THINKING QUESTION: Sy22000 nos quand i ydd Why do we use the embryonic cells (such...
CRITICAL THINKING QUESTION: Sy22000 nos quand i ydd Why do we use the embryonic cells (such as blastula cells) of animals to study mitosis? What cells would you use if you were studying mitosis in plants? Why? bonim. Do dalla di ni oldizi son 2 u n i ni oldiniz. . . Szargon ni
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Why do we want to avoid mixing the cells with water and heat fixing during the...
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3. A) Why is it necessary to include ampicillin in the medium with the (-) plasmid treatment? B) of the (+) plasmid treatment? the two genes of interest on the plasmid were only expressed on the plate with ampicillin. Why? 15. What causes the "halo effect", the white satellite colonies around the green fluorescent colonies, in the (+) plasmid TSA + ampicillin plate? (6. Other than transformation, what are two other mechanisms that bacteria use to acquire new genes?
Question 1 Not yet answered Marked out of 1.00 Flag question If we do a "negative control transformation experiment with no plasmid DNA added, we would expect to see Select one: a. no colonies on our transformation plates b. the same number of colonies as we see with the plasmid DNA added c. more colonies compared to when we add the plasmid DNA QuJLIUI Answer saved Marked out of 1.00 Flag question If we used the wrong antibiotic (kanamycin instead...
2. Why do broad-spectrum antibiotics such as streptomycin, chloramphenicol, and tetracycline inhibit a wide variety of prokaryotes, but do not harm human cells? a. 3. If an infectious agent is sensitive to several antimicrobial drugs, what other considerations might be taken into consideration to determine which drug should be prescribed to the patient? a 4. Look at a household cleanser or antiseptic that you use in your own home. What claims are made about its effectiveness against microbes (read the...
CRITICAL THINKING QUESTION: Sy22000 nos quand i ydd Why do we use the embryonic cells (such as blastula cells) of animals to study mitosis? What cells would you use if you were studying mitosis in plants? Why? bonim. Do dalla di ni oldizi son 2 u n i ni oldiniz. . . Szargon ni
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