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Which enzyme would cut the human DNA shown in Part A on both sides of the vgp gene but not inside the gene?
1. Which enzyme(s) would cut the human DNA shown in Part A on both sides of the vgp gene, but not inside the gene? Answer: BamHI, HaeIII, and HindIII 2. Which enzymes(s) would cut the plasmid without disrupting the function of the amp^R gene? Answer: BamHI, EcoRI, and HaeIII 3. Which enzyme(s) would produce sticky ends when cutting both the human DNA and the plasmid? Answer: BamHI, EcoRI, and HindIII 4. Which one restriction enzyme satisfies all three of the requirements listed above? Answer: BamHI only
EcoRI and HindIII are What?
They are type II restriction enzymes with a ommon structural core comprising four β-sheets and a single α-helix. restriction endonucleases
WHAT IS pBR322?
pBR322 is one of the most used cloning vectors in molecular biology. Cloning vectors, best-known as plasmids, are autonomously replicating DNA units into which DNA fragments can be inserted for gene cloning. Genes taken up by these plasmids are multiplied (or cloned) as the vector replicates, to yields numbers suitable for molecular analysis. The most versatile and well-known plasmid is certainly pBR322 (in fact was one of the first ever used in gene cloning techniques) and has genetically tailored cutting sites into which DNA can be inserted without affecting plasmid self-replication. pBR322 general characteristics are: a) Size: 4.3 kb; b) Replicon: ColE1, relaxed; c) Selective markers (resistance): Amp and Tet; d) Single sites (enzymatic restriction single sites): Ava I, Pst I, BamHI, PvuII, ClaI, SalI, EcoRI, and HindIII.
What is a DNA restriction site?
The restriction site is a sequence of DNA that is recognized by an endonuclease, or a protein that cuts DNA, as a site at which the DNA is to be cut. This cutting happens when restriction enzyme cleaves nucleotides by hydrolyzing the phosphodiester bond between them.
Function of hindiii?
Haemophilus influenzae is a species of bacteria that is the source of the HindIII restriction enzyme that cleaves the palindromic DNA sequence 5'-AAGCTT-3' in the presence of the cofactor Mg2+ via hydrolysis. While restriction enzymes cleave at specific DNA sequences, they are first required to bind non-specifically with the DNA backbone before localizing to the restriction site. On average, the restriction enzyme will form 15-20 hydrogen bonds with the bases of the recognition sequence. With the aid of other Van der Waals interactions, this bonding facilitates a conformational change of the DNA-enzyme complex which leads to the activation of catalytic centers. Despite the lack of evidence suggesting an exact mechanism for the cleavage of DNA by HindIII, site-mutagenesis analysis coupled with more detailed studies of metal ion-mediated catalysis in EcoRV have led to the following proposed catalytic mechanism. It has been suggested that during the hydrolysis of DNA by EcoRV the catalytic residue Lys-92 stabilizes and orients the attacking water nucleophile, while the carboxylate of Asp-90 stabilizes the leaving hydroxide anion through to coordination of Mg2
Which enzyme would cut the human DNA shown in Part A on both sides of the vgp gene but not inside the gene?
1. Which enzyme(s) would cut the human DNA shown in Part A on both sides of the vgp gene, but not inside the gene? Answer: BamHI, HaeIII, and HindIII 2. Which enzymes(s) would cut the plasmid without disrupting the function of the amp^R gene? Answer: BamHI, EcoRI, and HaeIII 3. Which enzyme(s) would produce sticky ends when cutting both the human DNA and the plasmid? Answer: BamHI, EcoRI, and HindIII 4. Which one restriction enzyme satisfies all three of the requirements listed above? Answer: BamHI only
What is a degradative enzyme that recognizes specific nucleotide sequences and cuts up DNA?
Those are restriction enzymes, a kind of endonucleases. Some wellknown ones are EcoRI, HindIII or BamHI
How restriction enzyms are use in DNA recommination research?
they used to cut the DNA at the specific site. For example: BamHI is a restriction enzyme that cuts between the given recognition site:
How many lambda DNA fragments does BamHI make?
There are five BamHI cut sites in lambda DNA: Location 5505 22346 27972 34499 41732 So, BamHI will digest lambda DNA into six fragments.
Why use bam h1 ans sau3a1 in plasmid transformation?
Because they share compatible sites. Whatever you digest with Sau3A1, you can ligate it in a BamHI site.
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
EcoRI and HindIII are What?
They are type II restriction enzymes with a ommon structural core comprising four β-sheets and a single α-helix. restriction endonucleases
WHAT IS pBR322?
pBR322 is one of the most used cloning vectors in molecular biology. Cloning vectors, best-known as plasmids, are autonomously replicating DNA units into which DNA fragments can be inserted for gene cloning. Genes taken up by these plasmids are multiplied (or cloned) as the vector replicates, to yields numbers suitable for molecular analysis. The most versatile and well-known plasmid is certainly pBR322 (in fact was one of the first ever used in gene cloning techniques) and has genetically tailored cutting sites into which DNA can be inserted without affecting plasmid self-replication. pBR322 general characteristics are: a) Size: 4.3 kb; b) Replicon: ColE1, relaxed; c) Selective markers (resistance): Amp and Tet; d) Single sites (enzymatic restriction single sites): Ava I, Pst I, BamHI, PvuII, ClaI, SalI, EcoRI, and HindIII.
What site provides good deals on cutting discs?
One can find cutting discs in many homeware stores, such as B&Q. Alternatively, one may try other stores such as Wickes, or webpage stores such as Ebay for cutting discs.
What is a DNA restriction site?
The restriction site is a sequence of DNA that is recognized by an endonuclease, or a protein that cuts DNA, as a site at which the DNA is to be cut. This cutting happens when restriction enzyme cleaves nucleotides by hydrolyzing the phosphodiester bond between them.
How many cords of hard wood can a athletic male cut in one day?
About one cord is the answer if you are felling the trees, limbing them, cutting them into firewood size pieces, loading them at the cutting site and unloading them at the delivery site. This assumes you can get your truck and trailer very close to the trees so no log skidding is required.
Function of hindiii?
Haemophilus influenzae is a species of bacteria that is the source of the HindIII restriction enzyme that cleaves the palindromic DNA sequence 5'-AAGCTT-3' in the presence of the cofactor Mg2+ via hydrolysis. While restriction enzymes cleave at specific DNA sequences, they are first required to bind non-specifically with the DNA backbone before localizing to the restriction site. On average, the restriction enzyme will form 15-20 hydrogen bonds with the bases of the recognition sequence. With the aid of other Van der Waals interactions, this bonding facilitates a conformational change of the DNA-enzyme complex which leads to the activation of catalytic centers. Despite the lack of evidence suggesting an exact mechanism for the cleavage of DNA by HindIII, site-mutagenesis analysis coupled with more detailed studies of metal ion-mediated catalysis in EcoRV have led to the following proposed catalytic mechanism. It has been suggested that during the hydrolysis of DNA by EcoRV the catalytic residue Lys-92 stabilizes and orients the attacking water nucleophile, while the carboxylate of Asp-90 stabilizes the leaving hydroxide anion through to coordination of Mg2
