The document discusses yeast molecular biology and genetics. It provides information on commonly used yeast species Saccharomyces cerevisiae and Schizosaccharomyces pombe. It discusses yeast genome, life cycle, vectors, cloning, transformation methods, and applications including gene expression and making mutants.
pOnebyOne™ are efficient, accurate and flexible Bicistronic Mammalian Expression Kits that contains an Expression Cassette based in 2A sequence breakthrough technology.
Its novel (patent pending) technology allows simultaneous Expression of two Proteins from the same mRNA. Cells transfected with Bicistronic vectors ensure that if one of the Proteins is present, the other one is also present.
Bicistronic Expression vectors are supported on viral elements: the IRES or 2A sequence. IRES has been widely used. It is a relative short sequence, around 600-700 bp, although this length could be a disadvantage in viral vectors where packaging capacity is limited. IRES based Expression vectors are characterized by a non-stoichiometric production of both proteins; generally there is a lower expression of the downstream gene.
Many 2A sequences from several families of viruses have been described for producing multiple polypeptides. 2A mediated cleavage is a universal phenomenon in all eukaryotic cells. With just 20 bp in length, the 2A sequence has been used succesfully to generate multiple proteins in some biological models: plants, zebrafish, transgenic mice or eukaryotic cell lines. Vectors based on 2A produce stoichiometric proportion of both proteins.
Canvax™ offers a ready-to-clone solution of your Gene of Interest, obtained by PCR, onto a wide collection of Bicistronic vectors based on 2A sequence. You can choose among different Promoters, selection Antibiotics or Reporter Genes.
this is a presentation on gene expression vector that includes what is expression vector, how many types of expression vector and difference between cloning and expression vector
Canvax™ offers a wide range of high quality Human Recombinant Proteins for several research applications like ELISA, Western Blot, Antibody Production or Protein array.
This PPT has described how to produce soluble anf high amount of recombinant protein in E.coli host. This PPT has mentioned different expression vectors, different E.coli Expression host strain and other strategies for getting high expression of desired gene.
pOnebyOne™ are efficient, accurate and flexible Bicistronic Mammalian Expression Kits that contains an Expression Cassette based in 2A sequence breakthrough technology.
Its novel (patent pending) technology allows simultaneous Expression of two Proteins from the same mRNA. Cells transfected with Bicistronic vectors ensure that if one of the Proteins is present, the other one is also present.
Bicistronic Expression vectors are supported on viral elements: the IRES or 2A sequence. IRES has been widely used. It is a relative short sequence, around 600-700 bp, although this length could be a disadvantage in viral vectors where packaging capacity is limited. IRES based Expression vectors are characterized by a non-stoichiometric production of both proteins; generally there is a lower expression of the downstream gene.
Many 2A sequences from several families of viruses have been described for producing multiple polypeptides. 2A mediated cleavage is a universal phenomenon in all eukaryotic cells. With just 20 bp in length, the 2A sequence has been used succesfully to generate multiple proteins in some biological models: plants, zebrafish, transgenic mice or eukaryotic cell lines. Vectors based on 2A produce stoichiometric proportion of both proteins.
Canvax™ offers a ready-to-clone solution of your Gene of Interest, obtained by PCR, onto a wide collection of Bicistronic vectors based on 2A sequence. You can choose among different Promoters, selection Antibiotics or Reporter Genes.
this is a presentation on gene expression vector that includes what is expression vector, how many types of expression vector and difference between cloning and expression vector
Canvax™ offers a wide range of high quality Human Recombinant Proteins for several research applications like ELISA, Western Blot, Antibody Production or Protein array.
This PPT has described how to produce soluble anf high amount of recombinant protein in E.coli host. This PPT has mentioned different expression vectors, different E.coli Expression host strain and other strategies for getting high expression of desired gene.
Rice is the principal food crop for more than half of the
world's population. Rice, as a staple food, supports more
than three billion people and comprises 50%–80% of their
daily calorie intake [1]. Adverse environmental factors
such as excessive cold, heat, drought, and salinity stresses
result in a considerable yield loss of crop plants all over
the world. Plant adaptations to environmental stresses
depend on the activation of cascades of molecularnetworks involved in signal transduction, stress perception,
and expressions of stress‐related genes. These
abiotic stresses elicit complex cellular responses in the
plant system, resulting in the production of excessive
reactive oxygen species (ROS) such as hydrogen peroxide
(H2O2), hydroxyperoxyl (HO2·), superoxide (O2
−), and
singlet oxygen (1O2) radicals. To protect themselves from
adverse conditions, plants have evolved a number of
cellular defense mechanisms including antioxidants such
as ascorbate, glutathione, and tocopherols as well as
ROS‐detoxifying enzymes such as superoxide dismutases
(SODs), peroxidases, and catalases (CATs) [2,3].
The content is about the general description of genetic material and further two techniques of biotechnology. The content includes two topics.
Firstly with introduction to biotechnology it describe about DNA, recombinant DNA (rDNA) technology, history, goals, procedure of rDNA technology, tools, techniques, application, demerits and products of rDNA technology.
Second portion entitiled as Hybridoma technology. this includes the basic principle, production of monoclonal antibodies, merits demerits and drugs from monoclonal antibody.
New pharmaceuticals derived from biotechnology is covered in last. All the content is referred from books and internet sources.
Generation of MRP2 Efflux Transporter Knock-Out in HepaRG Cell Linemdmitc
MilliporeSigma's Jennifer Pratt recently presented a poster at the 2016 AAPS/ITC Transporter Workshop demonstrating the utility of HepaRG MRP2 Knockout cells for investigating drug-transporter interactions in the liver involving MRP2.
Transcription factors of the nuclear factor κ B family are the paradigm for signaling dependent nuclear translocation and are ideally suited to analysis through image-based chemical genetic screening. The authors describe combining high-content image analysis with a compound screen to identify compounds affecting either nuclear import or export. Validation in silico and in vitro determined an EC50 for the nuclear export blocker leptomycin B of 2.4 ng/mL (4.4 nM). The method demonstrated high selectivity (Z′ >0.95), speed, and robustness in a screen of a compound collection. It identified the IκB protein kinase inhibitor BAY 11 7082 as an import inhibitor, the p38 mitogen-activated protein (MAP) kinase inhibitor PD98509 as an import enhancer, and phorbol ester as an export inhibitor. The results establish a robust method for identifying compounds regulating nucleocy- toplasmic import or export and also implicate MAP kinases in nuclear import of nuclear factor κ B
Driving school,Driving school in mentone,Driving Instructor Melbourne,mentone driving school,Melbourne driving schools,Driving school in clayton,Driving school in Dandenong,Driving license,Driving school in glenhuntly
LAST Conference - The Mickey Mouse model of leadership for software delivery ...Nish Mahanty
Leading an agile team can be rewarding and also challenging. It is an opportunity to apply your leadership and vision, and to introduce those the ideas and behaviours that are important to you. One of the main benefits is the opportunity to grow and develop the careers of your teams, and to have an impact wider than your own individual technical skills.
It is also a challenge. Often the skills that got you the promotion, or new job, aren't the ones you need to be successful in the new role. If you are inheriting an existing team, they usually have work in-flight so it’s important to be up to speed with what the team is doing, and whether they are on track for meeting their (now your) objectives. Every team, company, and situation is different, with unique challenges so it is important that you quickly identify where to focus your energies.
I'll outline a framework (with themes and a checklist) for assessing the situation, and constructing a 30 day plan to set yourself, and the team, up for success:
Theme 1: Build the things right (The technical aspects of delivering quality solutions)
Theme 2. Build the right thing (validating the planned deliverables against the desired business outcomes)
Theme 3. Build the right Team (building a resilient, highly engaged, highly skilled team, who work well together and who can efficiently adjust to unforseen changes, whilst still delivering the outcomes)
I believe that a successful agile team achieves a conscious balance between these themes. If they aren't focussed on all three, then they are unlikely to be as successful as they could be.
Against these three themes I'll present and discuss a 6 point checklist that will help the new leader develop a 30 day plan:
1. Business objectives and environment – assess whether the team is doing productive work that aligns with the business needs.
2. Team – build a highly engaged, resilient team that understand their contribution to the larger business outcomes
3. Metrics –continually visualise progress against your goals
4. Stakeholders – build a strong relationship, and clear lines of communication
5. Continual improvement – no team should stand still and no team has reached perfection, so continuously analyse performance and focus on getting better.
6. Budget – understand the financial commitment to help plan activities and team dynamics
The aim of the talk is to be educational, offering up a set of ideas, supported with real-world examples, that the attendees can adopt in their own organisations, to help them and their teams become more successful.
Rice is the principal food crop for more than half of the
world's population. Rice, as a staple food, supports more
than three billion people and comprises 50%–80% of their
daily calorie intake [1]. Adverse environmental factors
such as excessive cold, heat, drought, and salinity stresses
result in a considerable yield loss of crop plants all over
the world. Plant adaptations to environmental stresses
depend on the activation of cascades of molecularnetworks involved in signal transduction, stress perception,
and expressions of stress‐related genes. These
abiotic stresses elicit complex cellular responses in the
plant system, resulting in the production of excessive
reactive oxygen species (ROS) such as hydrogen peroxide
(H2O2), hydroxyperoxyl (HO2·), superoxide (O2
−), and
singlet oxygen (1O2) radicals. To protect themselves from
adverse conditions, plants have evolved a number of
cellular defense mechanisms including antioxidants such
as ascorbate, glutathione, and tocopherols as well as
ROS‐detoxifying enzymes such as superoxide dismutases
(SODs), peroxidases, and catalases (CATs) [2,3].
The content is about the general description of genetic material and further two techniques of biotechnology. The content includes two topics.
Firstly with introduction to biotechnology it describe about DNA, recombinant DNA (rDNA) technology, history, goals, procedure of rDNA technology, tools, techniques, application, demerits and products of rDNA technology.
Second portion entitiled as Hybridoma technology. this includes the basic principle, production of monoclonal antibodies, merits demerits and drugs from monoclonal antibody.
New pharmaceuticals derived from biotechnology is covered in last. All the content is referred from books and internet sources.
Generation of MRP2 Efflux Transporter Knock-Out in HepaRG Cell Linemdmitc
MilliporeSigma's Jennifer Pratt recently presented a poster at the 2016 AAPS/ITC Transporter Workshop demonstrating the utility of HepaRG MRP2 Knockout cells for investigating drug-transporter interactions in the liver involving MRP2.
Transcription factors of the nuclear factor κ B family are the paradigm for signaling dependent nuclear translocation and are ideally suited to analysis through image-based chemical genetic screening. The authors describe combining high-content image analysis with a compound screen to identify compounds affecting either nuclear import or export. Validation in silico and in vitro determined an EC50 for the nuclear export blocker leptomycin B of 2.4 ng/mL (4.4 nM). The method demonstrated high selectivity (Z′ >0.95), speed, and robustness in a screen of a compound collection. It identified the IκB protein kinase inhibitor BAY 11 7082 as an import inhibitor, the p38 mitogen-activated protein (MAP) kinase inhibitor PD98509 as an import enhancer, and phorbol ester as an export inhibitor. The results establish a robust method for identifying compounds regulating nucleocy- toplasmic import or export and also implicate MAP kinases in nuclear import of nuclear factor κ B
Driving school,Driving school in mentone,Driving Instructor Melbourne,mentone driving school,Melbourne driving schools,Driving school in clayton,Driving school in Dandenong,Driving license,Driving school in glenhuntly
LAST Conference - The Mickey Mouse model of leadership for software delivery ...Nish Mahanty
Leading an agile team can be rewarding and also challenging. It is an opportunity to apply your leadership and vision, and to introduce those the ideas and behaviours that are important to you. One of the main benefits is the opportunity to grow and develop the careers of your teams, and to have an impact wider than your own individual technical skills.
It is also a challenge. Often the skills that got you the promotion, or new job, aren't the ones you need to be successful in the new role. If you are inheriting an existing team, they usually have work in-flight so it’s important to be up to speed with what the team is doing, and whether they are on track for meeting their (now your) objectives. Every team, company, and situation is different, with unique challenges so it is important that you quickly identify where to focus your energies.
I'll outline a framework (with themes and a checklist) for assessing the situation, and constructing a 30 day plan to set yourself, and the team, up for success:
Theme 1: Build the things right (The technical aspects of delivering quality solutions)
Theme 2. Build the right thing (validating the planned deliverables against the desired business outcomes)
Theme 3. Build the right Team (building a resilient, highly engaged, highly skilled team, who work well together and who can efficiently adjust to unforseen changes, whilst still delivering the outcomes)
I believe that a successful agile team achieves a conscious balance between these themes. If they aren't focussed on all three, then they are unlikely to be as successful as they could be.
Against these three themes I'll present and discuss a 6 point checklist that will help the new leader develop a 30 day plan:
1. Business objectives and environment – assess whether the team is doing productive work that aligns with the business needs.
2. Team – build a highly engaged, resilient team that understand their contribution to the larger business outcomes
3. Metrics –continually visualise progress against your goals
4. Stakeholders – build a strong relationship, and clear lines of communication
5. Continual improvement – no team should stand still and no team has reached perfection, so continuously analyse performance and focus on getting better.
6. Budget – understand the financial commitment to help plan activities and team dynamics
The aim of the talk is to be educational, offering up a set of ideas, supported with real-world examples, that the attendees can adopt in their own organisations, to help them and their teams become more successful.
How can we include all businesses in the public sector to become digitally involved? Learn the journey we anticipate for a digital government and the benefits this can have on interaction and civil communications.
Thought Leadership from Social Sector MastersSalesforce.org
Some of the nation’s leading CRM thought leaders joined together for a lively and entertaining discussion about top marketing and CRM trends and best practices. This presentation is geared for social sector practitioners looking to take top lessons from different industries and apply them to their organization or campus. Tap over 75 years of combined experience on strategies to make customer initiatives successful from these featured thought leaders.
Google's recent announcement that it will support the use of microformats in their search opens up new possibilities for librarians and library technologists to support the goals of the semantic web; namely to provide better access, reuse and recombinations of library resources and services on the open web. This lightning talk introduces the semantic web and semantic markup technologies.
Lo Mejor de Cibeles Madrid Fashion Week - Otoño/Invierno 2010 - 2011Compulsiva Accesorios
Lo mejor de los mejores diseñadores (bajo mi criterio) de la Pasarela Cibeles para este Otoño/Invierno 2010 - 2011.
Un resumen bastante completo de las nuevas tendencias e ideas para el próximo invierno, y para "oler" hacia donde va el mercado.
Miss Compulsiva
This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
Lab: Differential Expression Differential gene expression provides the ability for a cell or
organism to respond to a constantly changing external environment. The specific constellation of
proteins required for optimal function and growth varies with cellular age and environmental
context. Thus, protein production is carefully regulated by multiple mechanisms that modulate
both transcriptional and translational pathways. Control of transcription initiation by RNA
polymerase is a predominant mechanism for regulating expression of specific proteins,
presumably because it provides maximal conservation of energy for the cell. We can often
observe the consequence of differential transcription due to the presence or absence of particular
proteins or the growth in particular environments. Control can also occur at translation; the
mRNA is synthesized, but only in certain circumstances is it translated. Control can also occur at
the level of protein function; the protein is inactive, or activity is not observed due to the lack of
the substrate. In this lab we will observe differential expression of two different genes encoded
on plasmids. We will analyze transcriptional activity, translational activity, and protein function.
Plasmids are extra-chromosomal DNA. Bacteria often have plasmids and will replicate the
plasmid and pass it to daughter cells (vertical transmission) and to neighboring cells (horizontal).
Plasmids are a mechanism of gene diversity. In order to stably retain the plasmid, there needs to
be some type of metabolic reason for the bacteria to maintain the plasmid. In other words, the
plasmid confers an advantage. Plasmids contain: 1. Ori: the plasmid may present is low or high
copy number. 2. Lab generated plasmids typically also contain a selectable marker (antibiotic
resistance), 3. Additional gene for ease of visual screening 4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant plasmids. The transformed cells containing the plasmid with the gene of interest ca.
Lab: Differential Expression Differential gene expression provides the ability for a cell or
organism to respond to a constantly changing external environment. The specific constellation of
proteins required for optimal function and growth varies with cellular age and environmental
context. Thus, protein production is carefully regulated by multiple mechanisms that modulate
both transcriptional and translational pathways. Control of transcription initiation by RNA
polymerase is a predominant mechanism for regulating expression of specific proteins,
presumably because it provides maximal conservation of energy for the cell. We can often
observe the consequence of differential transcription due to the presence or absence of particular
proteins or the growth in particular environments. Control can also occur at translation; the
mRNA is synthesized, but only in certain circumstances is it translated. Control can also occur at
the level of protein function; the protein is inactive, or activity is not observed due to the lack of
the substrate. In this lab we will observe differential expression of two different genes encoded
on plasmids. We will analyze transcriptional activity, translational activity, and protein function.
Plasmids are extra-chromosomal DNA. Bacteria often have plasmids and will replicate the
plasmid and pass it to daughter cells (vertical transmission) and to neighboring cells (horizontal).
Plasmids are a mechanism of gene diversity. In order to stably retain the plasmid, there needs to
be some type of metabolic reason for the bacteria to maintain the plasmid. In other words, the
plasmid confers an advantage. Plasmids contain: 1. Ori: the plasmid may present is low or high
copy number. 2. Lab generated plasmids typically also contain a selectable marker (antibiotic
resistance), 3. Additional gene for ease of visual screening 4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant plasmids. The transformed cells containing the plasmid with the gene of interest ca.
DNA damage repair Neil3 gene Knockout in MOLT-4iosrjce
RNAi is superannuated cellular mechanism that protect organism against viruses that replicate
through double- stranded RNA. RNAi can be used to diminish gene expression from plasmid expressing and
inserted sequence repeat. A stable harpin would be expressed after the vector was integrated into the genome.
In this paper a shiRNA expressing vector for Neil3 was designed and developed which is capable of replication
in MOLT-4. This shiRNA vector had the ability to arose the RNAi pathway, and reduce the gene expression of
Neil3. This was assessed by using pSilence 4.1CMV as a vector, and Gapdh as positive control.
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE B.pdfamzonknr
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE
BACKGROUND CONTEXT Lab: Differential Expression Differential gene expression provides
the ability for a cell or organism to respond to a constantly changing external environment. The
specific constellation of proteins required for optimal function and growth varies with cellular
age and environmental context. Thus, protein production is carefully regulated by multiple
mechanisms that modulate both transcriptional and translational pathways. Control of
transcription initiation by RNA polymerase is a predominant mechanism for regulating
expression of specific proteins, presumably because it provides maximal conservation of energy
for the cell. We can often observe the consequence of differential transcription due to the
presence or absence of particular proteins or the growth in particular environments. Control can
also occur at translation; the mRNA is synthesized, but only in certain circumstances is it
translated. Control can also occur at the level of protein function; the protein is inactive, or
activity is not observed due to the lack of the substrate. In this lab we will observe differential
expression of two different genes encoded on plasmids. We will analyze transcriptional activity,
translational activity, and protein function. Plasmids are extra-chromosomal DNA. Bacteria often
have plasmids and will replicate the plasmid and pass it to daughter cells (vertical transmission)
and to neighboring cells (horizontal). Plasmids are a mechanism of gene diversity. In order to
stably retain the plasmid, there needs to be some type of metabolic reason for the bacteria to
maintain the plasmid. In other words, the plasmid confers an advantage. Plasmids contain: 1. Ori:
the plasmid may present is low or high copy number. 2. Lab generated plasmids typically also
contain a selectable marker (antibiotic resistance), 3. Additional gene for ease of visual screening
4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant.
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE BAC.pdfamzonknr
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE
BACKGROUND CONTEXT Lab: Differential Expression Differential gene expression provides
the ability for a cell or organism to respond to a constantly changing external environment. The
specific constellation of proteins required for optimal function and growth varies with cellular
age and environmental context. Thus, protein production is carefully regulated by multiple
mechanisms that modulate both transcriptional and translational pathways. Control of
transcription initiation by RNA polymerase is a predominant mechanism for regulating
expression of specific proteins, presumably because it provides maximal conservation of energy
for the cell. We can often observe the consequence of differential transcription due to the
presence or absence of particular proteins or the growth in particular environments. Control can
also occur at translation; the mRNA is synthesized, but only in certain circumstances is it
translated. Control can also occur at the level of protein function; the protein is inactive, or
activity is not observed due to the lack of the substrate. In this lab we will observe differential
expression of two different genes encoded on plasmids. We will analyze transcriptional activity,
translational activity, and protein function. Plasmids are extra-chromosomal DNA. Bacteria often
have plasmids and will replicate the plasmid and pass it to daughter cells (vertical transmission)
and to neighboring cells (horizontal). Plasmids are a mechanism of gene diversity. In order to
stably retain the plasmid, there needs to be some type of metabolic reason for the bacteria to
maintain the plasmid. In other words, the plasmid confers an advantage. Plasmids contain: 1. Ori:
the plasmid may present is low or high copy number. 2. Lab generated plasmids typically also
contain a selectable marker (antibiotic resistance), 3. Additional gene for ease of visual screening
4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant.
A transplastomic plant is a genetically modified plant in which the new genes have not been inserted in the nuclear DNA but in the DNA of the chloroplasts.
Plasmids are small, circular pieces of DNA that can be found in prokaryotic cells. They can be naturally occurring or artificially created, and they can carry genes that are not found in the host cell's genome. Plasmids can be used to express genes in prokaryotes in a number of ways.
One way to express a plasmid-borne gene is to use a promoter. A promoter is a sequence of DNA that binds to RNA polymerase, the enzyme that initiates transcription. When RNA polymerase binds to a promoter, it begins to transcribe the gene, which produces a messenger RNA (mRNA) molecule. The mRNA is then translated into a protein by ribosomes.
Another way to express a plasmid-borne gene is to use a transcription factor. A transcription factor is a protein that binds to a specific sequence of DNA and regulates the transcription of genes. When a transcription factor binds to a DNA sequence, it can either increase or decrease the rate of transcription of the gene.
Plasmid-borne gene expression can be used for a variety of purposes, including:
Producing proteins for research or industrial purposes
Engineering bacteria to produce new products or to perform new functions
Developing vaccines or other medical treatments
Plasmid-borne gene expression is a powerful tool that can be used to manipulate the genetic makeup of prokaryotic cells. This technology has the potential to revolutionize the way we produce food, develop new medicines, and protect the environment.
Here are some additional points about plasmid borne gene expression in prokaryotes:
The copy number of a plasmid can affect the level of gene expression.
The host cell's environment can also affect the level of gene expression.
Plasmids can be used to transfer genes between different prokaryotic cells.
Plasmids can be used to create new strains of bacteria with desired properties.
gene cloning in eukaryotes (gene transfer).pdfNetHelix
Gene cloning recently faced difficulties associated with
bacteria, especially when dealing with
genes from eukaryotic organisms so we should to employ the eukaryotic expression in this PDF we will learn about gene cloning in eukaryotes, types of yeast plasmids and the importance of each one
systems
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
19. URA3 : The gene product of URA3 (orotidine-5’-phosphate decarboxylase) converts 5-FOA (5-fluoroorotic acid) to a toxic product that kills the URA3 cells. LYS2 : The LYS2 gene encodes a-aminoadipate reductase, an enzyme required for lysine biosynthesis. Yeast cells with wild-type LYS2 activity will not grow on media containing - aminoadipate ( -AA) as a primary nitrogen source. CAN1 : The CAN1 gene encodes an arginine permease. In the absence of arginine, canavanine (arginine analog) is readily incorporated into proteins with lethal consequences; therefore, CAN1 cells are sensitive to canavanine. CYH2 : The CYH2 gene encodes the L29 protein of the yeast ribosome. Cycloheximide blocks translation elongation by interacting with L29. Yeast negative selection systems: