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What enzyme is produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18?
The enzyme produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18 is luciferase. This enzyme is responsible for the bioluminescent properties of animals like fireflies and glowworms. Cells transformed with plasmid lux will emit light in the presence of the substrate luciferin, whereas cells transformed with pUC18 will not.
Why do the cells transformed with pUC18 and plasmid lux grow in the presence of ampicillin?
The pUC18 plasmid contains the ampicillin resistance gene (ampR) which confers resistance to ampicillin. The Lux operon on the plasmid allows for bioluminescence production and acts as a reporter gene. Therefore, transformed cells that harbor both plasmids can grow in the presence of ampicillin due to pUC18 and express bioluminescence due to the Lux operon.
What is pSC in vector pSC101?
Stanley Cohen, who constructed the plasmid
Is a cloning vector a plasmid?
plasmid is the type of the cloning vector. other cloning vectors includes cosmids, bacteriophage, phagemids, artifiical chromosomes. clonong vectors are the carriers of certain traits to be inserted in non coding regions of the DNA.
How can foreign pieces of DNA be transfered into organisms?
By a vector such as a plasmid or by a BAC or YAC.
What is the abbreviation of puc18?
puc18- plasmid of university of california 18
What enzyme is produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18?
The enzyme produced by cells transformed with plasmid lux that is not produced by cells transformed with pUC18 is luciferase. This enzyme is responsible for the bioluminescent properties of animals like fireflies and glowworms. Cells transformed with plasmid lux will emit light in the presence of the substrate luciferin, whereas cells transformed with pUC18 will not.
Why do the cells transformed with pUC18 and plasmid lux grow in the presence of ampicillin?
The pUC18 plasmid contains the ampicillin resistance gene (ampR) which confers resistance to ampicillin. The Lux operon on the plasmid allows for bioluminescence production and acts as a reporter gene. Therefore, transformed cells that harbor both plasmids can grow in the presence of ampicillin due to pUC18 and express bioluminescence due to the Lux operon.
What is pSC in vector pSC101?
Stanley Cohen, who constructed the plasmid
What would happen if you cut both the jellyfish glo gene and puc18 plasmid with the ecor1 restriction enzyme?
If there is a EcoR1 site in either the middle of the Glo gene, or in the middle of the selectable marker site in the plasmid, it would likely disable either Glo, or the plasmid.
Is a cloning vector a plasmid?
plasmid is the type of the cloning vector. other cloning vectors includes cosmids, bacteriophage, phagemids, artifiical chromosomes. clonong vectors are the carriers of certain traits to be inserted in non coding regions of the DNA.
Is a plasmid a cloning vector?
Yes, a plasmid can be used as a cloning vector. Plasmids are small, circular DNA molecules that can replicate independently in a host cell. They can carry foreign DNA fragments and be used to introduce these fragments into host cells for gene cloning and expression.
You have propagated a vector in a non expression host and isolated the plasmid How can you confirm whether the plasmid that you got is exactly the vector which you propagated?
Check the size in agarose gel, extract it from the gel then purify it and grow it on selective plate.
Significance of vector in DNA recombinant technology?
Vector are plasmid DNA, act as a molecular vehicles to carry genes or DNA of interest. In rDNA technology vectors used to clone the gene by ligation. This chimeric DNA or plasmid can be propagated in E.coli as the vector carries its own origin of replication. Expression plasmid vectors can be used to produce proteins from the gene of interest.
How can foreign pieces of DNA be transfered into organisms?
By a vector such as a plasmid or by a BAC or YAC.
How do you express a human gene in E. coli?
E.coli is used to express human genes because it can be easily grown in the lab. The gene is extracted from the DNA (by doing a partial digest with a restriction enzyme), and given a cohesive sticky end with a linker or adapter. It is then ligated to a plasmid vector, which had a restriction site compatible with the ends on the gene, eg if the plasmid contains a BamH1 site then you would add a linker or adapter which is compatible with the 5'GGATCC3' BamH1 recognition sequence. The cells are transformed (made to take up the plasmid vector) by chemical treatment; they are mixed with the plasmid, then a strong concentration of calcium (Ca2+) ions is added to the mixture to make the E.coli's membranes porous. The mixture is then heated to heat-shock the cells, to approx 50 degrees C for one minute. They are then cooled and allowed to recover in a nutrient rich broth at optimum temperature. This is a very inefficient process - only about 1 cell in every million is transformed. The pUC18 plasmid vector is useful because it contains the gene for ampicillin resistance. Any cells which subsequently grow on a medium containing ampicillin, therefore, have been transformed with the plasmid vector. It is also useful because it contains a beta-galactosidase gene, which itself contains the recognition site for a number of restriction enzymes, including BamH1. This is good because you can tell if the vector has taken up the gene you are trying to express when the vector no longer codes for the beta-galactosidase protein product. If the vector has been ligated with the gene, the gene will have disrupted the beta-galactosidase gene. This can be tested with IPTG (an auto-inducer) and X-gal, which will turn colonies of E.coli with the beta-galactosidase gene intact blue (ie, those without the gene of interest). Colonies which have had their beta-galactosidase gene destroyed by the ligation of the gene of interest will be colourless in the presence of X-gal and IPTG. These colonies are all clonal, so all cells in colourless colonies contain copies of the pUC18 plasmid vector which has been ligated with the human gene.
What is the difference between Puc18 and Puc19?
The multiple cloning sites (MCS) in pUC18 and pUC19 is the difference - the MCS in pUC19 is reverse orientated to those of the pUC18.pUC18: LacZ HindIII PaeI ..... SacI EcoRIpUC19: Lacz EcoRI SacI ..... PaeI HindIII
