The document describes methods for producing recombinant proteins and transgenic organisms. It discusses using genetic engineering techniques like inserting exogenous DNA into organisms like plants, animals and microbes. The DNA is then expressed to produce therapeutic proteins like insulin, growth hormone, erythropoietin or modify traits in crops and livestock. The methods include identifying and isolating genes, inserting them into vectors, transforming cells and regenerating organisms or culturing microbes to produce the protein of interest.
2. Golden rice
The Golden Rice was developed and patented in 2000 by
the public scientists Ingo Potrykus and Peter Beyer.
Golden rice is a variety of rice produced through genetic
engineering to biosynthesize beta-carotene, a precursor of
vitamin A, in the edible parts of rice. It is intended to
produce a fortified food to be grown and consumed in
areas with a shortage of vitamin A.
3. Method Of Preparation Of Golden Rice
The psy and crtI genes were transferred into the rice nucleus and
placed under the control of an endosperm-specific promoter.
The exogenous lcy gene has a transit peptide sequence attached, so it
is targeted to the plastid, where geranylgeranyl diphosphate is
formed.
The bacterial crtI gene was an important inclusion to complete the
pathway, since it can catalyze multiple steps in the synthesis of
carotenoids up to lycopene, while these steps require more than one
enzyme in plants.
The end product of the engineered pathway is lycopene, but if the
plant accumulated lycopene, the rice would be red. Recent analysis
has shown the plant's endogenous enzymes process the lycopene to
beta-carotene in the endosperm, giving the rice the distinctive yellow
colour for which it is named
4.
5. Production Of Recombinant FACTOR VIII
This application is a continuation of U.S. application Ser.
No. 13/000,938, filed on Dec. 22, 2010
Factor VIII is an essential blood-clotting protein, also
known as anti-hemophilic factor (AHF). In humans, factor
VIII is encoded by the F8 gene. Defects in this gene result
in hemophilia A, a recessive X-linked coagulation disorder.
Factor VIII is produced in liver sinusoidal cells
6. METHOD OF PREPERATION OF recombinant
FACTOR VIII
Isolate mature mRNA (containing only exon and no
introns) that is responsible for synthesis for factor viii.
This mRNA contains 9000 bases synthesis the protein,
factor viii.
Now synthesis the complementary DNA (cDNA) for mature
mRNA of factor viii.
This cDNA can be inserted into mammalian cells or
hamster kidney cells for production of Recombinant
FACTOR VIII.
7.
8. Production of recombinant erythropoietin
This is U.S. patent application Ser. No. 675,298, filed on
Nov. 30, 1984 and issued as U.S. Pat. No. 4,703,008 on Oct.
27, 1987
The introduction of recombinant human erythropoietin has
revolutionized the treatment of patients with anemia of
chronic renal disease. Clinical studies have explained that
recombinant human erythropoietin is also useful in various
non-uremic conditions including hematological and
oncological disorders, prematurity, HIV infection, and
perioperative therapies.
9. Method of preparation of recombinant
erythropoietin
Isolate and Construct human Epos cDNA.
Subjecting the cDNA to PCR using primer based on published
sequence.
The PCR Product will be cloned into vector for purpose of
propagation and subsequently engineered into appropriate expression
vector.
Culture -production purification.
10.
11. Process for the introduction of exogenous
DNA in somatic and germ animal cells
This application is European patent of international
application Ser. No. WO 1990008192A filed on Jan. 26,
1990 under Patent Cooperation Treaty.
A process is described for the introduction of exogenous
DNA into somatic and germ animal cells: the DNA, by
process of recombinant DNA it is introduced into the
animal spermatozoa which are to be modified and said
spermatozoa are employed for egg fertilization according
to usual artificial fertilization techniques.
12. Method of introduction of exogenous
DNA in somatic and germ animal cells
a) preparation of an aqueous spermatozoa suspension
b) transformation of the spermatozoa with cloned DNA
c) "in vitro" fertilization of the oocytes by means of the modified
spermatozoa
d) implantation of the fertilized oocytes into pseudo gravid females of the
selected species.
According to another possible way of performing the present invention, the
modified spermatozoa may be employed directly for the animal fertilization
without going through the "in vitro" oocyte fertilization and successive
implantation of the same into pseudo gravid females.
13.
14. Method for preparing transgenic animal
This application is European patent of international
application Serial No. WO97/11597 filed on 30.09.1998.
A method for preparing a sperm containing an exogenous
DNA, wherein the exogenous DNA together with liposome
for the purpose of transduction of DNA into mammalian
cells is injected repeating three times or more into a
testis of a mature non-human vertebrate male and the
sperm containing the exogenous DNA is produced in the
testis of said male.
15. Method of preparing transgenic animal
an exogenous DNA is added to a solution containing liposome, mixed and
incubated.
A liposome-exogenous DNA complex is directly injected into a testis of
mature non-human vertebrate male.
The number of injections is carried out preferably three times or more. The
mature males are anesthetized and the solution containing liposome
described above is injected directly from the scrotum into both testes.
The injection procedure is repeated at four-day intervals for at least three
times. On day 2 from the final injection of the liposome-exogenous DNA
complex, a sperm containing the exogenous DNA is produced in the testis of
said male.
16. Forming transgenic animal
A transgenic animal can be prepared by mating said males
with females of whom estruses has been induced by
gonadotropic hormone, making the females pregnant and
delivered.
A transgenic animal can be prepared by artificial
insemination
In-vitro fertilization using a sperm produced by the test is
of a mature non-human vertebrate male that has been
subjected to intratesticular injection of the exogenous
DNA and by making the females pregnant and delivered
17.
18. Process of producing oil from algae using
biological rupturing
This is U.S. patent application Ser. No. 60/877,786, filed
on Dec. 29, 2006 which is incorporated by reference
herein in its entirety.
The present invention relates generally to conversion of
algae and other biomass to biofuels such as biodiesel or
bioethanol.
19. Method of preparing oil from algae using
biological rupturing
In this invention by using algae to produce lipids (oil) which can be readily
converted into biodiesel through rupturing the cell wall and oil vesicles of the
algae.
In accordance with one aspect, a process for production of biofuels from
algae can include cultivating an oil-producing alga, extracting the algal oil,
and converting the algal oil to form biodiesel.
Extracting the algal oil from the oil-producing algae can include biologically
rupturing cell wall and oil vesicles of the oil-producing algae using at least
one cellulase, glycoproteins, cellulose, virus, or combination thereof.
Additionally, a system for production of biodiesel from algae can include algae
growth reservoirs, an oil extraction bioreactor connected to the growth
reservoir, a biological agent source connected to the oil extraction bioreactor,
and a conversion reactor operatively connected to the oil extraction
bioreactor.
20.
21. Method for development of transgenic
goats
This application is European patent of international
application Ser. No. WO 97/19589 filed on June.5,1997 by
NEXIA Biotechnologies.
The method of the invention preferably entails expansion
and propagation of dwarf goat progeny.
22. Method of preparing transgenic goats
introducing a transgene into a zygote of a dwarf goat,
transplanting the zygote into a pseudo pregnant non-dwarf goat,
allowing the zygote to develop to term. In another aspect the
invention features a method which includes the following steps: -
introducing a transgene into an embryo of a dwarf goat,
transplanting the embryo into a pseudo pregnant non-dwarf goat,
and allowing the embryo to develop to term.
23.
24. Gene therapy
This application is European patent of international
application Serial No. 220,175 filed on March. 30,1994
This invention relates to the use of primary human cells as
vehicles for human gene transfer. More particularly, this
invention relates to the use of human cells (such as, for
example, but not limited to, human blood cells) as
vehicles for the transfer of human genes encoding
therapeutic agents and/or genes encoding detectable
markers.
25. METHOD
Gene therapy is a technique that modifies a person’s genes to treat or
cure disease. Gene therapies can work by several mechanisms:
Replacing a disease-causing gene with a healthy copy of the gene
Inactivating a disease-causing gene that is not functioning properly
Introducing a new or modified gene into the body to help treat a
disease
Gene therapy products are being studied to treat diseases including
cancer, genetic diseases, and infectious diseases.
26.
27. Preparation method of transgenic plants
This application is European patent of international
application Serial No. WO 00/15813 filed on March. 23,2000
GM is a technology that involves inserting DNA into the
genome of an organism. To produce a GM plant, new DNA is
transferred into plant cells. Usually, the cells are then
grown in tissue culture where they develop into plants. The
seeds produced by these plants will inherit the new DNA.
28. METHOD
Genetic engineering is the modification of an organism's phenotype by
manipulating its genetic material. Some genetic engineering uses the
principle of recombination.
Recombination is the process through which a new gene is inserted into a
bacterial DNA "The plasmid".
The DNA needs to be cut with an enzyme called a restriction enzyme. The
restriction enzyme used must have a specific shape that allows it to move
along the DNA that is to be cut. The restriction enzyme looks for a
specific point in the DNA sequence at which to cut the DNA.
When the restriction enzyme cuts, it leaves a "Sticky end" which helps a
new gene to attach at that point. Another enzyme is used to attach the
new DNA segment; this is called "DNA ligase“
. Genetically engineered bacterium is cultured and many new copies of
the bacteria with the new gene are grown.
Genetic modifications can be made to both plants and animals.
29.
30. Process for production of recombinant
human growth hormone from E. coli cells
This is a U.S. National Phase application under 35 U.S.C. of
International Patent Application No. PCT/IB2011/002348, filed on Sep.
20, 2011, and claims the priority of European Patent Application No.
10177997.3, filed Sep. 21, 2010 both of which are incorporated by
reference herein in their entirety. The International Application
published in English on Mar. 29, 2012 as WO 2012/038822 A1 under
PCT Article 21
Human growth hormone (h GH), also known as somatropin (INN) or
somatotropin, is a protein hormone produced and secreted by the
somatotropic cells of the anterior pituitary. Human growth hormone
plays a key role in somatic growth in childhood and in metabolism in
adulthood through its effects on the metabolism of proteins,
carbohydrates and lipids.
31. Method
The gene for human growth hormone (h GH) is isolated
from human pituitary gland.
Insertion of whole h GH gene into plasmid vector and
cloning into E. coli results into production of biologically
inactive hormone because bacteria can translate the
region of gene that are not translated in human thereby
producing a pre hormone containing an extra 26 amino
acids which might be difficult to remove.
Hence the segment of gene that codes for the first 24
amino acids of hormone is constructed chemically from
blocks of nucleotide.
32.
33. Method for producing human recombinant insulin
This application is European patent of international application Serial
No. PCT/UA2009/000025 filed on 16.06.2009
Insulin has been used for many years to treat diabetes. Diabetes is
well managed by taking insulin. Earlier insulin was extracted from the
pancreas of killed cattle and pigs. It had shortcomings. It used to
stimulate allergic reactions and other immune responses due to its
foreign origin in some people. Another challenge was to cater to the
ever-increasing demand and large-scale production.
To overcome this, the production of insulin by recombinant DNA
technology was done and it has proved to be very beneficial. In fact,
it was the first recombinant medicine to be used in the USA.
Biosynthetic insulin produced by rDNA technology is purer than animal
insulin. It reduces the formation of antibodies against it.
34. Method for producing human recombinant insulin
Insulin is a protein hormone synthesized by the 𝛃 cells of the pancreas.
It is produced as a prohormone, i.e., PR proinsulin. The signal peptide cleaves
to give proinsulin. Proinsulin has to be further processed to become functional.
Proinsulin contains another peptide chain known as ‘C’ peptide in addition to ‘A’
and ‘B’ peptides, which are required for its functionality. This C peptide is not
present in mature insulin. The main challenge for production of insulin using r
DNA technique was getting insulin assembled into mature form. This two DNA
sequence corresponding A and B, chains of human insulin and introduced them in
plasmid of E. coli to produce insulin chains. Chain A and chain B were produced
separately. Chain A and Chain B were produced separately extracted and
combined by creatin disulfide bond to form human insulin.
From proinsulin, the C peptide is removed at the time of maturation.
The genetically engineered insulin does not contain the C peptide
35.
36. NEW HAIR BOTOX MATERIAL
This application is United States Patent of application
Serial No. US8,795,643B1 filed on Aug. 5, 2014
This invention includes an improvement on hair Botox.
The invention is concerned with the development of a
new hair care and repair material particularly with hair
Botox and a new method of application of this material to
hair. A much longer lasting and more effective hair care
and repair operation can be realized through the
invention in question.
37. METHOD
In the method in which the keratin is used in liquid form, when keratin
protein (the one used in the invention is hydrolyzed keratin) is applied
directly to the damaged parts of the hair,
the keratin proteins are enabled to be dry after the evaporation of the water
that provides the solution, and they unite with the hair like bone, repair and
fill the hair surface, making the hair smooth and shiny.
In addition, these benefits, which it brings for the hair, will never lose later
unlike as in synthetic and chemical materials or other methods.
The basic feature of the subject of the invention, the hair Botox, is to mix
water with keratin in certain rates and to apply it directly to the hair.
