Answer: The given experiment is related to the bacterial transformation. Bacterial transformation is the transfer of foreign genetic material (either DNA or RNA) into the recipient cell through direct uptake from the environment. It is a process involves exchange of genetic material between of two organisms through non-reproductive mode in contrast to the traditional reproduction (vertical transfer through sexual or asexual reproduction). It results to the change in the genotype of the recipient cell due to incorporation of newly acquired DNA into the recipient genome through either recombination or insertion. The transformants can be identified using a selective medium. The identification of the transformants (containing the vector/foreign molecule) require the presence of a suitable marker genes on the vector molecule, whose expression provides a means of identifying cells containing it (transformants). The most popular marker used for it are: a) Antibiotic resistant gene, b) colour substance developing gene, c) galactosidase gene complementation. In the given experiment 3 markers were used: a) The vector (pGLO plasmid) contain the gene for Green Fluorescent Protein (GFP) and the ampicillin resistance gene (which codes for beta-lactamase). Because it shares a bidirectional promoter with a gene for metabolizing arabinose, the GFP gene is expressed in the presence of arabinose, which makes the transgenic organism express its fluorescence under UV light. GFP can be induced in bacteria containing the pGLO plasmid by growing them on +arabinose plates. b) Ampicillin antibiotic, inhibits the cell wall synthesis. It acts as a competitive inhibitor of the enzyme transpeptidase. c) Arabinose (as a carbon source for bacteria). The genetic transformation resulting to the change the phenotype of the organism (E. coli in case of the given experiment). The results found after the transformation experiment would be in observed in 4 type of plates. 1) Control plate (Growth medium only) 2) Control plate (Growth medium+ampicillin) 3) Transformant plate (Growth medium+ampicillin+pGLO) 4) Transformant plate (Growth medium+ampicillin+pGLO+arabinose) The observations of control plate (E. coli cells before transformantion experiment) might provide reference data to determine if any genetic transformation has occurred. Thus, we have to record the a) Number of colonies, b) Size of the colony, c) Color of the colonies, d) Distribution of the colonies on the plate, e) Visible appearance when viewed with ultraviolet (UV) light), f) The ability of the cells to survive and multiply in the presence of an antibiotic ampicillin. Expected Results: *Plate 1: The results would obtained that countable (between 30-300), small, circular, shiny, colourless colonies of E. coli were distributed throught out the plate. They would be non- fluorescent when obsered under UV-light. *Plate 2: No colonies will be developed in the presence of ampicillin because the E. coli is non resistant .

1. Answer:
The given experiment is related to the bacterial transformation. Bacterial transformation is the
transfer of foreign genetic material (either DNA or RNA) into the recipient cell through direct
uptake from the environment. It is a process involves exchange of genetic material between of
two organisms through non-reproductive mode in contrast to the traditional reproduction
(vertical transfer through sexual or asexual reproduction). It results to the change in the genotype
of the recipient cell due to incorporation of newly acquired DNA into the recipient genome
through either recombination or insertion.
The transformants can be identified using a selective medium. The identification of the
transformants (containing the vector/foreign molecule) require the presence of a suitable marker
genes on the vector molecule, whose expression provides a means of identifying cells containing
it (transformants). The most popular marker used for it are: a) Antibiotic resistant gene, b) colour
substance developing gene, c) galactosidase gene complementation.
In the given experiment 3 markers were used:
a) The vector (pGLO plasmid) contain the gene for Green Fluorescent Protein (GFP) and the
ampicillin resistance gene (which codes for beta-lactamase). Because it shares a bidirectional
promoter with a gene for metabolizing arabinose, the GFP gene is expressed in the presence of
arabinose, which makes the transgenic organism express its fluorescence under UV light. GFP
can be induced in bacteria containing the pGLO plasmid by growing them on +arabinose plates.
b) Ampicillin antibiotic, inhibits the cell wall synthesis. It acts as a competitive inhibitor of the
enzyme transpeptidase.
c) Arabinose (as a carbon source for bacteria).
The genetic transformation resulting to the change the phenotype of the organism (E. coli in case
of the given experiment). The results found after the transformation experiment would be in
observed in 4 type of plates.
1) Control plate (Growth medium only)
2) Control plate (Growth medium+ampicillin)
3) Transformant plate (Growth medium+ampicillin+pGLO)
4) Transformant plate (Growth medium+ampicillin+pGLO+arabinose)
The observations of control plate (E. coli cells before transformantion experiment) might provide
reference data to determine if any genetic transformation has occurred. Thus, we have to record
the a) Number of colonies, b) Size of the colony, c) Color of the colonies, d) Distribution of the
colonies on the plate, e) Visible appearance when viewed with ultraviolet (UV) light), f) The
ability of the cells to survive and multiply in the presence of an antibiotic ampicillin.
Expected Results:

2. *Plate 1: The results would obtained that countable (between 30-300), small, circular, shiny,
colourless colonies of E. coli were distributed throught out the plate. They would be non-
fluorescent when obsered under UV-light.
*Plate 2: No colonies will be developed in the presence of ampicillin because the E. coli is non
resistant to ampicillin antibiotic.
*Plate 3: The results would obtained that some relatively smaller, and some larger circular,
shiny, colourless colonies of E. coli were found in the plate. These are ampicillin resistant
transformants of E. coli. They would be non-fluorescent when obsered under UV-light because
the GFP gene will not expressed in the absence of arabinose.
*Plate 4: The results would obtained that two types of colonies of E. coli were found in the plate,
some are green coloured transformants colonies and some are non-transformant (non-
fluorescent) but ampicillin resistant colonies. The fluorescent colonies (when obsered under UV-
light) are transformant colonies of E. coli, because the GFP gene is expressed in the presence of
arabinose. Thus, the genetic transformation causing the alteration in the gene of the E. coli
(ampicillin sensitive to ampicillin resistant) and change in the phenotype of the cells (colourless
to green colour colonies).
Solution
Answer:
The given experiment is related to the bacterial transformation. Bacterial transformation is the
transfer of foreign genetic material (either DNA or RNA) into the recipient cell through direct
uptake from the environment. It is a process involves exchange of genetic material between of
two organisms through non-reproductive mode in contrast to the traditional reproduction
(vertical transfer through sexual or asexual reproduction). It results to the change in the genotype
of the recipient cell due to incorporation of newly acquired DNA into the recipient genome
through either recombination or insertion.
The transformants can be identified using a selective medium. The identification of the
transformants (containing the vector/foreign molecule) require the presence of a suitable marker
genes on the vector molecule, whose expression provides a means of identifying cells containing
it (transformants). The most popular marker used for it are: a) Antibiotic resistant gene, b) colour
substance developing gene, c) galactosidase gene complementation.
In the given experiment 3 markers were used:
a) The vector (pGLO plasmid) contain the gene for Green Fluorescent Protein (GFP) and the
ampicillin resistance gene (which codes for beta-lactamase). Because it shares a bidirectional
promoter with a gene for metabolizing arabinose, the GFP gene is expressed in the presence of

3. arabinose, which makes the transgenic organism express its fluorescence under UV light. GFP
can be induced in bacteria containing the pGLO plasmid by growing them on +arabinose plates.
b) Ampicillin antibiotic, inhibits the cell wall synthesis. It acts as a competitive inhibitor of the
enzyme transpeptidase.
c) Arabinose (as a carbon source for bacteria).
The genetic transformation resulting to the change the phenotype of the organism (E. coli in case
of the given experiment). The results found after the transformation experiment would be in
observed in 4 type of plates.
1) Control plate (Growth medium only)
2) Control plate (Growth medium+ampicillin)
3) Transformant plate (Growth medium+ampicillin+pGLO)
4) Transformant plate (Growth medium+ampicillin+pGLO+arabinose)
The observations of control plate (E. coli cells before transformantion experiment) might provide
reference data to determine if any genetic transformation has occurred. Thus, we have to record
the a) Number of colonies, b) Size of the colony, c) Color of the colonies, d) Distribution of the
colonies on the plate, e) Visible appearance when viewed with ultraviolet (UV) light), f) The
ability of the cells to survive and multiply in the presence of an antibiotic ampicillin.
Expected Results:
*Plate 1: The results would obtained that countable (between 30-300), small, circular, shiny,
colourless colonies of E. coli were distributed throught out the plate. They would be non-
fluorescent when obsered under UV-light.
*Plate 2: No colonies will be developed in the presence of ampicillin because the E. coli is non
resistant to ampicillin antibiotic.
*Plate 3: The results would obtained that some relatively smaller, and some larger circular,
shiny, colourless colonies of E. coli were found in the plate. These are ampicillin resistant
transformants of E. coli. They would be non-fluorescent when obsered under UV-light because
the GFP gene will not expressed in the absence of arabinose.
*Plate 4: The results would obtained that two types of colonies of E. coli were found in the plate,
some are green coloured transformants colonies and some are non-transformant (non-
fluorescent) but ampicillin resistant colonies. The fluorescent colonies (when obsered under UV-
light) are transformant colonies of E. coli, because the GFP gene is expressed in the presence of
arabinose. Thus, the genetic transformation causing the alteration in the gene of the E. coli
(ampicillin sensitive to ampicillin resistant) and change in the phenotype of the cells (colourless
to green colour colonies).