Download Chapter IV – Microbial Cell Culture and its Applications.

Winter break holiday home work
BIOTECHNOLOGY HOTS QUESTIONS.
Chapter IV – Microbial Cell Culture and its Applications.
1. A student tried to isolate insulin from recombinant E. coli cells. He collected the
filtrate and performed chromatography. However protein was not obtained. What
may be the possible reason?
2. A researcher isolated a fungus which can produce a secondary metabolic compound.
Suggest any method by which he can increase the yield of the product?
3. At NCBS Bangalore, the researchers isolated a new strain of bacteria, for further
studies they preserved it in mineral oil. What is the advantage of storing the slant in
mineral oil?
4. For performing a microbiology experiment Arjun sterilized the medium in Hot air
oven, but he couldn’t get the properly sterilized media. Why?
5. To avoid the growth of unnecessary microorganisms, while doing the experiment
Santhi added antibiotics to the medium. Suggest by which method can she sterilize
the antibiotics?
6.
If one wishes to obtain intracellular metabolites. What type of microbial culture
technique should be used?
7. Name the type of bacteria that does not use biotin on minimal media.
8. Name the first commercial genetically engineered protein?
9. In the Pathology laboratory of NCCS, Pune, the researchers isolated a group of
microorganisms from soil in a Petri plate. Name the culture technique that can be
used for the isolation of specific microorganism using a specially designed medium?
10. Name the most common type of continuous culture device that can control both
population density as well as the growth rate of the culture?
11. A student performing an investigatory project wanted to produce Penicillin. He
selected E.coli cells for this. However he dint get the expected result. What can be
the possible reason for this?
12. You are asked to design a bioreactor. What are the main features that you have to
keep in mind while designing it?
13. In batch culture the growth of the bacteria is inhibited due to high concentration of
the substrate. How can you avoid this?
14. The specific growth rate of a few bacteria is given below. From the data given find
out which microorganism has the least doubling time?
Microorganisms
µ max (h -1)
Benechea natriegens
Methylomonas methynolytica
Aspergillus nidulans
Penicillium chrysogenum
Chapter2 Recombinant DNA Technology
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1. If you wanted to express a eukaryotic protein in bacterial cells would you
clone genomic DNA or c-DNA in to the expression vector? Justify your
choice. (3 marks)
2. If a genetic engineer has to choose a vector in order to clone single stranded
desired DNA than which vector would you suggest.(2 marks)
3. (a)What technique is depicted in the below diagram?( 1+2+2 =5marks)
(b)Based on the information available from the autoradiograph write the
sequence of sample DNA.
(c)What is the role of ddNTPs in this technique?
4. The given DNA fragment is cleaved by EcoRI then it was separated by gel
electrophoresis. Represent the fragments we will get after electrophoresis. (3
marks)
EcoRI
EcoRI
1kb
4kb
3.5kb
5. The given sequence is:
a) 5’ GAATTCGATACATTAA 3’
b) 5’ TTACCTATGACTAGAA 3’
c) 5’ AATACTAGGATCCATA 3’
d) 5’ CATGACTAGCATAAT 3’
In which sequence BamHI restriction site is present.( 3marks)
6. Why there need to convert mRNA into cDNA to form a library instead of the
genomic DNA. (2 marks)
7. If you want to check the production of recombinant proteins which vector is useful
for this purpose and why? (3 marks)
8. If you have a very small quantity of DNA sample then which techniques can be
used for amplifies it? Also write the use of Mg2+ in this technique?(3 marks)
9. Which technique you can use to produce the desired mutation in the gene. Also
give the application of this technique. (3marks)
10. What is the use of Probe in southern hybridization? Is it necessary to make
the probe radio labeled or enzyme labelled, if yes then why? (3 marks)
11. If you have done the gene cloning by using vector PBR322 then which method would
be useful to identify the recombinants? (2marks)
12. If the sequence read from autoradiogram is 5’ GAATTCATACGTCACTG 3’ it will
appear on the gel.(3marks)
13. In Human Genome project why BAC vector is used instead of YAC.Why? (2marks)
14. If there are four strands of DNA sample, after 25 cycles of PCR, How
many copies of DNA will be produced? (3 marks)
15. Imagine yourself as a biotechnologist and you have to propogate a gene of your
intrest into bacteria and have to express it into Yeast which particular vector will
you use and why? (3 marks)
HOTS
CLASS XII
PLANT CELL CULTURE AND APPLICATION
16. Q1. Why the maintenance in aseptic environment is necessary for the growth of
2 cultured tissues?
17.
18. Q2. How will you sterilize the growth regulators and why?
2
19.
20. Q3. Why does the public view GM food with concerns? Indicate three possible
reasons.
2
21.
22. Q4. During the preparation of virus free plants from a virus infected cell, a plant
botanist used a differentiated plant cell but the experiment failed. Which cell is
to be used for a successful experiment and why?
3
23.
24. Q5. You have been asked by a avocado grower to find a way to genetically
engineer his crop to prevent it from ripening before shipping. What
experimental approaches you suggest.
3
25. Q6 When substance X is added to a plant tissue culture medium, it promoted
rooting whereas when substance Y is added,it promoted shooting. Identify
substances X and Y.
2
26.
27. Q7.A plant biotechnologist used Coptis. japonica for commercial production of
barberine, but he got very less amount of barberine and there was a heavy
money loss. Next time he analysed some limiting factors for failure for his
experiment and he got better results. Suggest those factors by justifying your
answer.
3
28.
29. Q.8 What do you mean by the opposite acting activity of Barstar and Barnase.
3
30.
31.
Q.9What do you mean by Shikonin, and what is its function.
2
32.
33.
Q.10What is the actual name of the macerozyme and what is its function .
34.
2
35.
Q.11 What do you mean by the co-cultivation of the protoplasts.
36.
3
37.
Q.12 Define the over all procedure of plant tissue culture,as shown in the following
picture. 3
38.
Protein structure and engineering
1- Which of the following amino acid substitution will be least harmful in primary structure
of a protein?
a- Arginine is replaced by lysine
b- Tryptophan is replaced by glutamate
c- Valine is replaced by aspartic acid
d- Glutamate is replaced by valine.
(1)
2- A protein is analyzed by mass spectrometry, which of the following amino acid
substitution in this protein is not going to affect the result.
A-Glycine is substituted by glutamine
B-Phenylalanine is substituted by methionine
C- Leucine by isoleucine
D-Aspartic acid by glutamic acid
(1)
3-Can naked protein causes disease? Give your critical comments.
(2)
4-Why large amount of protein is needed by Sanger for protein sequencing. (3)
5-Disulfide bond in a protein always represent primary structure of protein .Give the critical
comment.
(3)
6- Why the deficiency of ADA causes SCID?
(3)
7-What type of secondary structure is present in collagen?
(1)
8-Why the absorbance of the proteins is always measured at 280 nm?
(2)
9- How the serine residue in some enzyme can became acidic and also suggest that how
can you confirm that a serine residue is involved in catalysis.
(3)
10- If a protein is incorrectly folded than what is the consequence? Give an example to
illustrate your answer.
(2)
11-Why Branched chain amino acids are important for the muscle growth and activity?
(2)
12-Genes code for proteins but how only 30000 genes of the human beings are able to form
90000 of proteins present in our body. Explain?
(3)
13- As we know AUG is start codon and it codes for the methionine. So the first amino acid
of every protein is always methionine. Give your critical comments.
14- A mixture of five proteins A,B,C,D,E with molecular mass of 20,50,43,55 and 3
kilodalton respectively is subjected to gel permeation chromatography.
(3)
(a) Which protein will elute first from the column.
(b) Which protein will elute last from the column.
(c) What is another name of gel permeation chromatography?
15-(a) Why humulin is preferred over the insulin obtained from the pancreas of cow and
pig.
(2+3)
(b) Study the following enzyme purification table and answer the question that follows
Step
Procedure
1
2
3
Total protein
(mg)
1200
300
150
Crude extract
Precipitation(salt)
Ion exchange
chromatography
4
Gel chromatography
120
5
Affinity chromatography
3
a) Which step in the purification is least effective? Why?
b) Which step in the purification is most effective? Why?
c) Which is the yield of active protein from crude extract?
Activity
(units)
2400
2100
1800
1600
1200
GENOMICS AND BIOINFORMATICS
1. Name the search engine used to connect these given databases. 1
2. Find out the no. of amino acids possible from the given m-RNA sequence sequences.
AUGAGUAGCAACACCUAA.
1
3. i) What is the difference between these two sequences given below?
ii) Name the variation.
……GGTAACTG……
2
……GGCAACTG……
4. Identify the regions of a gene in the given picture.
2
5. Write two differences between EST and C-DNA
6. Name two uses of the given database.
2
2
7.
Databases
……?………
PDB
PALI
………?………
8.
9.
information available
Nucleotide sequence
……………?………….
…………?……………..
r-RNA subunit sequence
2
i)Identify the disease given in the picture
ii) Mention the chromosomal phenomenon happening here.
iii) Name the technique by which this chromosomal phenomenon can be identify.
3
a UACGACUUU b
I I I I I I I I I III I I I
Study the RNA segment given above which is completely translated into
A polypeptide chain.
3
(i) Write codon a and b.
(ii) What do they code for?
(iii) How is peptide bond formed between two amino acids in the
Ribosome?
10. Name the major steps involved in a comparative micro array experiment.
5
11. In-silico gene prediction is not very accurate give at least three reasons supporting the
statement.
3
12. If you are given a sequence without any label, How will you find out whether it is a DNA
sequence or a RNA sequence or a Protein sequence.
2
13. If a Scientist isolate an unknown gene he wants to go for homology search, What steps
he should follow?
5
HOTS
PLANT CELL CULTURE AND APPLICATIONS
Q1. Why the maintenance in aseptic environment is necessary for the growth of cultured
tissues?
Q2. How will you sterilize the growth regulators and why?
Q3. Why does the public view GM food with concerns?Indicate three possible reasons.
Q4. During the preparation of virus free plants from a virus infected cell, a plant botanist
used a differentiated plant cell but the experiment faile.Which cell is to be used for a
successful experiment and why?
Q5. You have been asked by a avocado grower to find a way to genetically engineer his
crop to prevent it from ripening before shipping. What experimental approaches you
suggest.
Q6 When substance X is added to a plant tissue culture medium, it promoted rooting
whereas when substance Y is added,it promoted shooting. Identify substances X and Y.
Q7.A plant biotechnologist used Coptis. japonica for commercial production of barberine,
but he got very less amount of barberine and there was a heavy money loss. Next time he
analysed some limiting factors for failure for his experiment and he got better
results.Suggest those factors by justifying your answer.