A.

The researchers extracted the vector in piggyback transposons, and the genes of
interest are GFP from the jellyfish Aequorea victoria and human insulin.

B. The restriction enzyme used to cut is the EcoRI.

C. Polymerase Chain Reaction They applied the laboratory technique known as

"pcr," which makes millions to billions of copies of a certain DNA region very
quickly. Additionally, they chose the genome to be amplified using DNA
fragments known as primers before doing numerous rounds of DNA synthesis.
Polymerase chain reaction has a big work in doing the gmo, which is applied a lot
of number of DNA molecules.

D. To find the GFP transgene in mouse DNA, the PCR (polymerase chain reaction)
method is frequently utilized. A specific DNA sequence can be amplified using
PCR, which can produce millions of copies of the target DNA from a little quantity
of starting material. The GFP transgene in this instance is the target sequence.
After that, gel electrophoresis can be used to see the PCR products and compare
them to a positive control (i.e., a plasmid containing the GFP gene). Fluorescence
microscopy is another method used to identify transgenic mice expressing the GFP
transgene. GFP emits a green fluorescent light when exposed to certain
wavelengths of light. Therefore, tissues or cells from the transgenic mice can be
examined under a fluorescence microscope to detect the green fluorescence
signal. This method allows researchers to visualize the expression pattern of the
GFP transgene in living cells or tissues.

E. It plays an important role in preserving genomic integrity by joining breaks in the

phosphodiester backbone of DNA that happen during replication and
recombination.

F. Microinjection is generally used to introduce the GFP gene-containing vector into

mouse embryos. Using this method, the vector is directly injected into the
pronucleus of a freshly fertilized mouse embryo, which houses the haploid genetic
material from both the egg and the sperm. The pseudopregnant female mice are
used as surrogate mothers for the gestation of the microinjected embryos after they
are implanted into their oviducts. The sterile men are mated with females to
produce pseudopregnant females, which results in a hormonal condition that
resembles pregnancy. After being surgically implanted into the oviducts of the
fictitious pregnant females, the microinjected embryos can grow into fully
developed mice.
 G. In general, hundreds of mouse stocks with mutations that have been long utilized
as models of human disease and the investigation of metabolic systems would
determine when GMO glow-in-the-dark mice would begin to proliferate. By
inserting foreign DNA into the mouse cell genome with increased specificity,
scientists have improved their ability to produce mutant strains. These transgenes
may be randomly inserted into the genome. The resultant mouse exhibits the trait
that the foreign DNA codes for. Alternately, mutations can be directed at certain
loci to create "knockout mice" that do not express the gene or to change the way
that gene expression is expressed.

REFERENCES:
 Ivics, Z., & Izsvák, Z. (2004). The expanding universe of transposon technologies for
gene and cell engineering, Mobile DNA, 1(1), 25 Grabundzija, I., Irgang, M., Mátés,
L., Belay, E., Matrai, J., Gogol-Döring, A.,... & Izsvák, Z. (2010). Comparative analysis
of transposable element vector systems in human cells, Molecular Therapy, 18(6),
1200–1209.
 youtube.com/watch?v=i4Ivu62t0t8
 https://www.genome.gov/genetics-glossary/Polymerase-Chain-
Reaction#:~:text=Polymerase%20chain%20reaction%20(abbreviated%20PCR,be%
20studied%20in%20greater%20detail.
 Zimmer, M. (2002) Green fluorescent protein (GFP): applications, structure, and
related photophysical behavior Chemical Reviews, 102 (3), 759–781. Baker, M.
(2012) Transgenic mice: The history, current state of the art, and future directions
Methods in Molecular Biology, 859, 1–16 Chalfie, M., & Kain, S. (2005). Green
fluorescent protein: properties, applications, and protocols John Wiley & Sons
 https://tinyurl.com/bdza848n
 Hogan, B., Beddington, R., Costantini, F., & Lacy, E. (1994). Manipulating the Mouse
Embryo: A Laboratory Manual (2nd ed.) Cold Spring Harbor Laboratory Press
Capecchi, M. R. (1980). High-efficiency transformation by direct microinjection of DNA
into cultured mammalian cells. Cell 22(2), 479–488 Nagy, A., Gertsenstein, M.,
Vintersten, K., & Behringer, R. (2003). Manipulating the Mouse Embryo: A Laboratory
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 https://www.ncbi.nlm.nih.gov/books/NBK231336/