I need help answering these 3 questions based on biology and plasmids.

1. Just mixing your insulin gene and plasmid DNA w/ sticky ends may temporarily result in a new plasmid, but it is being held together by weak hydrogen bonds. What enzyme do you need to add in order to make ur new plasmid a solid circle of DNA?

6. What is the purpose of using plasmids with antibiotic resistance in them?

7. Why were no restriction enzyme tested that would produce bunt ends instead of sticky ends?

Technorati Tags: antibiotic resistance, biology, bunt, hydrogen bonds, insulin gene, plasmid dna, restriction enzyme, sticky ends

i’ll give u the options

cellulose
heterocysts
hyphae
gills
spores

Cutting DNA with a particular restriction enzyme produces ______ that can be separated by gel electrophoresis.

plasmids
restriction fragments
recombinant DNA
enzymes
a genomic library

Technorati Tags: cellulose, dna enzymes, gel electrophoresis, genomic library, recombinant dna, restriction enzyme, restriction fragments, spores

I need help with a few questions:

1: Why did we make sure to include the start and stop DNA sequences for the Jellyfish Glo gene in our cut segment

2:What would have happened if we had cut both the Jellyfish Glo gene and puc18 plasmid
with the EcoR1 restriction enzyme? Be sure to look on the paper DNA sequences to find the
EcoR1 restriction enzyme cut sites.

3:If we want to now produce a lot of this Jellyfish Glo protein, what do we have to do after this
first successful cloning to reach our goal?

The Lab is at this link:

http://www.explorebiology.com/documents/21CloningSimulation.pdf

2:

Technorati Tags: cloning, dna sequences, jellyfish, plasmid, protein, restriction enzyme, segment

I’m trying to learn this stuff from a textbook (without a lab), so it’s all pretty abstract…

If I were to want to sequence an entire gene in an individual organism… could I just use PCR to amplify the gene and then sequence that with Sanger Sequencing? Presuming the gene is longer than than 800 nucleotides, is there a way to "primer walk" or "shotgun sequence" using solely the products of PCR?

Or, am I just supposed to clone fragments of the DNA using vectors, and then use those for the sequencing and primer walking?

If I am supposed to clone the fragments, then should I still use PCR to make the original fragment, and then chop THAT up with restriction enzymes to get the smaller fragments… or is PCR an unnecessary step in all this.

I’m kinda confused. Any help would be very much appreciated!

Thank you!!!
thank you all very much for your answers.

yutgoyun, if you’re still there, are you suggesting that I clone the entire gene in one plasmid? To do that, wouldn’t I still need to cut the gene with a restriction enzyme so that it could recombine with the plasmid?

please excuse my ignorance… this is all completely new to me.

thanks again.

Technorati Tags: dna, fragment, fragments, nucleotides, organism, plasmid, restriction enzyme, restriction enzymes, sanger sequencing, shotgun, textbook, unnecessary step, vectors

If you wanted to clone the soxA gene using restriction enzyme PvuI, would you be best to use to use pUC18 or pBR322 as a cloning vector?
would it be pBR322, since it has small size?? which can facilitates entry of the plasmid into cells?

Technorati Tags: cells, cloning vector, pbr322, plasmid, puc18, restriction enzyme

A scientist wants to insert a gene for vitamin A into a rice plant, the scientist would use ________.

a)a vector
b)a restriction enzyme
c)DNA ligase
d)All of the above

Technorati Tags: dna ligase, restriction enzyme, rice plant, scientist, vector

1. Which of the following proteins are not coded for by genes carried on plasmids?
a. enzymes necessary for conjugation
b. enzymes that catabolize hydrocarbons
c. bacteriocins
d. enzymes that inactivate antibiotics
e. none of the above
2. an enzyme produced in response to the presence of a substrate is called
a. an inducible enzyme
b. a repressible enzyme
c. a restriction enzyme
d. an operator
e. a promoter
3. conjugation differs from reproduction because conjugation
a. replicates DNA
b. transfers DNA vertically, to new cells
c transfers DNA horizontally, to cells in same generation
d transcribes DNA to RNA
e none of the above

Technorati Tags: antibiotics, bacteriocins, cells, conjugation, dna, enzymes, Genes, hydrocarbons, inducible enzyme, presence, promoter, proteins, replicates, reproduction, restriction enzyme, rna, substrate

A plasmid has two antibiotic resistance genes, one for ampicillin and one for tetracycline. It is treated with a restriction enzyme that cuts in the middle of the ampicillin gene. DNA fragments containing a human goblin gene were cut with the same enzyme. The plasmids and fragments are mixed, treated with ligase, and used to transform bacterial cells. Clones that have taken up the recombinant DNA are the ones that

A) are blue and can grow on plates with both antibiotics.

B) can grow on plates with ampicillin but not with tetracycline.

C) cannot grow with any antibiotics.

D) can grow on plates with tetracycline and are not blue.

Technorati Tags: ampicillin, antibiotic resistance genes, antibiotics, bacterial cells, clones, goblin, ligase, plasmid, Plasmids, recombinant dna, restriction enzyme, tetracycline

a plasmid has two antibiotic resistance genes, one for ampicillin and one for tetracycline. it is treated wit ha restriction enzyme that cuts in the middle of the ampicillin gene. DNA fragments containing a human globin gene were cut with the same enzyme. the plasmids and fragments are mixed, treated with ligase and used to transform bacterial cells. clones that have taken up the recombinant DNA are the ones that…
a. are blue and can grow on plates with both antibiotics
b. can grow on plates with ampicillin but not with tetracycline.
c. can grow on plates with tetracycline but not with ampicillin
d. cannot grow with any antibiotics
e. can grow on plates with tetracycline and are not blue

Technorati Tags: ampicillin, antibiotic resistance genes, antibiotics, bacterial cells, clones, globin gene, ligase, plasmid, Plasmids, recombinant dna, restriction enzyme, tetracycline, wit

a. the sequence of genes on a chromosome
b. the sequence regognized by a restriction enzyme
c. the sequence of bases in a DNA segment
d. the number of VNTR repeats in a chromosome section

2. To identify an individual by DNA analysis of their blood at crime scenes, investigators look for
a. primers
b. DNA profiles
c. probes
d. nucleosomes
e. transgenic fragments

3. What is the goal of PCR
a. intro. of DNA into reproductively cloned animals
b. amplification of small amounts of protein
c. seperation of DNA by size
d. making many copies of a transgenic animal
e. incrasing the amount of DNA when the starting smaple is small

4. which of the following is the most commoly used cloning vector in genetic engineering?
a. plasmids
b. restriction enzymes
c. bacteriophages
d. DNA
e. genomics

Technorati Tags: amplification, bacteriophages, chromosome, crime scenes, dna analysis, dna profiles, dna segment, Genes, genetic engineering, genomics, nucleosomes, Plasmids, primers, probes, protein c, restriction enzyme, restriction enzymes, seperation, transgenic animal, vntr