What is the
function of each gene seen in this plasmid?Homework Answers
Plasmid: Plasmid is a circular, double-stranded genetic material,that is distinct from a cell's chromosomal DNA. Plasmids naturally exist in bacterial cells, and they also occur in some eukaryotes.
Genes on Plasmid: The genes carried in plasmids provide bacteria with genetic advantages, such as antibiotic resistance
Function of each gene:
AmpR gene:The AmpR gene, which confers resistance to the antibiotic named ampicillin.
Example:E. coli cells containing this plasmid, termed "+ampR" cells, can survive and formcolonies on LB agar that has been supplemented with ampicillin.
PDC1:This gene is responsible in the production of Major of three pyruvate decarboxylase isozymes, key enzyme in alcoholic fermentation, decarboxylates pyruvate to acetaldehyde; subject to glucose-, ethanol-, and autoregulation; involved in amino acid catabolism.
Ori : This is the DNA sequence that signals for the origin of replication,process. some times this is referred to simply as origin. In E. coli, ori is some 250 nucleotides in length for the chromosomal origin (oriC). The plasmid orisequences are similar to oriC, and are called oriV (origin of vegetative replication).
ARS1: This contains the origin of replication in plasmids. It contains four regions (A, B1, B2, and B3), named in order of their effect on plasmid stability. The A-Domain is highly conserved, any mutation abolishes origin function. Mutations on B1, B2, and B3 will diminish, but not prevent functioning of the origin.
CEN4:CEN4 is the centromere which is useful for the segregation during the process of cerll division.
URA3: URA3 is often used in yeast research as a "marker gene", that is, a gene to label chromosomes or plasmids.
lacZ:A multiple cloning site (MCS) is present within the lacZsequence in the plasmid. This sequence can be nicked by restriction enzymes to insert the foreign DNA.
HindIII and BamHI are the Restriction enzyme present on the plasmid.
Add Answer to:
What is the
function of each gene seen in this plasmid?
Hindlll -0.03 BamHI-1.5 YOpDCI 12.8...
You have accidentally torn the labels off two tubes, each containing a different plasmid, and now...
You have accidentally torn the labels off two tubes, each
containing a different plasmid, and now do not know which plasmid
is in which tube. Fortunately, you have restriction maps for both
plasmids, shown in the figure below. You have the opportunity to
test just one sample from one of your tubes. You have equipment for
agarose-gel electrophoresis, a standard set of DNA size markers,
and the necessary restriction enzymes.
A)Why won’t making a single cut (e.g. with HindIII) help...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
2. (Multiple answers) A student is doing an experiment to insert the plasmid seen below into...
2. (Multiple answers) A student is doing an experiment to insert the plasmid seen below into an E. coli bacteria. They cut their plasmid and gene of interest with the HIND III restriction enzyme to insert their gene of interest. lacz HindIII BamHI -EcoRI amp PUC19 After the experiment is complete they observe many blue colonies on their plate media that contained nutrients, X-gal and ampicillin. Please choose all that are TRUE about this experiment. ampicillin is the selective ingredient...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb)....
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
You have cloned the Pepsi Gene into the vector pKEN. You used two restriction enzymes, BamHI...
You have cloned the Pepsi Gene into the vector pKEN. You used two restriction enzymes, BamHI and EcoRI to force the cloning in a directional way. The plasmid (pKEN) is ~5 kb long and the insert Pepsi 800bp long. After transformation into E. coli you grow up several colonies and isolate the vector. After digestion of the vector it looks like you have three potential clones. You need to sequence the gene to make sure you have the correct clone....
Question 2,3 and 4 aBbCcDdE AaBbCoDdE Normal No Spacing BamHI 300 EcoRI 3400 1000 2000 5 kb EcoRU Apal List three components a cloning vector must contain A cloning site A replication origin A sel...
Question 2,3 and 4
aBbCcDdE AaBbCoDdE Normal No Spacing BamHI 300 EcoRI 3400 1000 2000 5 kb EcoRU Apal List three components a cloning vector must contain A cloning site A replication origin A selectable marker -Drug resistant gene 1. 2. Draw the end of the fragment that will be generated after digestion with the Apal enzyme. Indicate the type of ends generated by this enzyme (Blunt/Sticky, 5' or 3' overhang) (3) 3. What is the size of the plasmid...
He full-length SNFI gene was inserted into pLexA at the BamHl restriction site to create pl.ex-Sn...
he full-length SNFI gene was inserted into pLexA at the BamHl restriction site to create pl.ex-Snf. To do this, the BamHI recognition sequence was introduced onto both the 5 and 3 ends of the SNFI DNA. Add the necessary nucleotides onto both strands of the DNA represented below: 5' 3 SNF1 sense strand SNF1 antisense strand 3 5 Show the sequence that will result from BamHl digestion: 5" 3 SNF1 sense strand SNF1 antisense strand 3' 5 2. What are...
4. During cloning we insert an ampicillin resistance gene a plasmid, (a) Explain its function in...
4. During cloning we insert an ampicillin resistance gene a plasmid, (a) Explain its function in the cloning experiment. (b) Sketch and explain what would occur if you plate your post-cloning specimens on (1) a plate with methicillin and (ii) ampicillin? Explain the results in your sketch.
Question 1 - 12 What does the AMPr gene in the PGEM-T plasmid produce and what...
Question 1 - 12 What does the AMPr gene in the PGEM-T plasmid produce and what is the role of this product in the protocol itself? Type your answer Question 2 --12 What does the lacZ gene in the PGEM- T plasmid produce and what is the role of this product in the protocol itself? Type your answer Question 3 (413) What occurs during the ligation step of Protocol 11? Type your answer Question 4 ( -- /2 What is...
Assume there is a plasmid, pAGG1 that carries two genes, geneA encoding the antibiotic resistance gene...
Assume there is a plasmid, pAGG1 that carries two genes, geneA encoding the antibiotic resistance gene to select the plasmid and geneX which is your gene of interest. Each of the two genes is about 0.6kb encoding proteins of about 200 amino acids. Question: Discuss what limitations there are in mutagenizing the entire plasmid to saturate geneX, when would you select a whole gene method to mutagenize geneX versus a site directed method?
You have accidentally torn the labels off two tubes, each
containing a different plasmid, and now do not know which plasmid
is in which tube. Fortunately, you have restriction maps for both
plasmids, shown in the figure below. You have the opportunity to
test just one sample from one of your tubes. You have equipment for
agarose-gel electrophoresis, a standard set of DNA size markers,
and the necessary restriction enzymes.
A)Why won’t making a single cut (e.g. with HindIII) help...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
2. (Multiple answers) A student is doing an experiment to insert the plasmid seen below into an E. coli bacteria. They cut their plasmid and gene of interest with the HIND III restriction enzyme to insert their gene of interest. lacz HindIII BamHI -EcoRI amp PUC19 After the experiment is complete they observe many blue colonies on their plate media that contained nutrients, X-gal and ampicillin. Please choose all that are TRUE about this experiment. ampicillin is the selective ingredient...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
Question 2,3 and 4
aBbCcDdE AaBbCoDdE Normal No Spacing BamHI 300 EcoRI 3400 1000 2000 5 kb EcoRU Apal List three components a cloning vector must contain A cloning site A replication origin A selectable marker -Drug resistant gene 1. 2. Draw the end of the fragment that will be generated after digestion with the Apal enzyme. Indicate the type of ends generated by this enzyme (Blunt/Sticky, 5' or 3' overhang) (3) 3. What is the size of the plasmid...
he full-length SNFI gene was inserted into pLexA at the BamHl restriction site to create pl.ex-Snf. To do this, the BamHI recognition sequence was introduced onto both the 5 and 3 ends of the SNFI DNA. Add the necessary nucleotides onto both strands of the DNA represented below: 5' 3 SNF1 sense strand SNF1 antisense strand 3 5 Show the sequence that will result from BamHl digestion: 5" 3 SNF1 sense strand SNF1 antisense strand 3' 5 2. What are...
4. During cloning we insert an ampicillin resistance gene a plasmid, (a) Explain its function in the cloning experiment. (b) Sketch and explain what would occur if you plate your post-cloning specimens on (1) a plate with methicillin and (ii) ampicillin? Explain the results in your sketch.
Question 1 - 12 What does the AMPr gene in the PGEM-T plasmid produce and what is the role of this product in the protocol itself? Type your answer Question 2 --12 What does the lacZ gene in the PGEM- T plasmid produce and what is the role of this product in the protocol itself? Type your answer Question 3 (413) What occurs during the ligation step of Protocol 11? Type your answer Question 4 ( -- /2 What is...
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