Biology 1202B Lecture Notes - Dominance (Genetics), Galactose, Mutagen

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ochrebee715

30 Apr 2012

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Biotechnology - technology applied to biological systems or living organisms to make or modify products or processes for a specific purpose: genetic engineering manipulation of genetic material. Bacterial restriction enzymes or restriction endonucleases (re: re recognize short amount of dna sequence (4-8nt long) called restriction sites, restriction sites are often palindromes - same sequence 5" to 3" on both strands. Incomplete digestion - conditions are such that if a restriction enzyme site is present in the dsdna, the enzyme may cut at all the dsdna sites. Isolate gdna and re cuts it into fragments. The same re cuts a bacterial plasmid. Mix the cut gdna with the cut plasmid (they have the same sticky ends) This creates recombinant dna, which is inserted into a bacterial cell. The bacteria grow and divide (replicates and amplifies the gdna) Selection - insertion of genes to identify which bacteria have a modified plasmid. Give plasmids ampicillin resistance gene, so it can live in ampicillin.

Related Questions

We have four DNA sequences that we need to digest (cut up), and have 6 restriction enzymes in stock in our lab. We want to find out which restriction enzyme we can use on each sample.

For each sequence, please indicate which restriction enzyme(s) would cut within each sequence. Each sequence may be cut by one, more than one, or no restriction enzymes.

Following the sequences are two questions. Please choose from the same list of enzymes to indicate the correct answer to these questions.

Restriction enzyme	Digestion pattern
EcoRI	5' G\|AATTC 3' 3' CTTAA\|G 5'
HindIII	5' A\|AGCTT 3' 3' TTCGA\|A 5'
BamHI	5' G\|GATCC 3' 3' CCTAG\|G 5
AluI	5' AG\|CT 3' 3' TC\|GA 5'
SmaI	5' CCC\|GGG 3' 3' GGG\|CCC 5'
HbaI	5' GCG\|C 3' 3' C\|GCG 5'

(The | symbol indicates the specific cut sites.)

Each answer may be used once, more than once, or not at all.

1.	5' -TAGACTGAATTCAAGTCA- 3' 3' -ATCTGACTTAAGTTCAGT- 5'
2.	5' -ATACGCCCGGGTTCTAAA- 3' 3' -TATGCGGGCCCAAGATTT- 5'
3.	5' -CAGGATCGAAGCTTATGC- 3' 3' -GTCCTAGCTTCGAATACG- 5'
4.	5' -AATAGAATTCCGATCCGA- 3' 3' -TTATCTTAAGGCTAGGCT- 5'
5.	Which restriction enzyme(s) is(are) 4-cutters (i.e. they recognize and cut at 4-bp sequences, instead of 6-bp sequences)?
6.	Which restriction enzyme(s) do not leave a "sticky end" after cutting the DNA?

A.	Alu
B.	AluI, SmaI
C.	All 6 of these restriction enzymes.
D.	None of the 6 restriction enzymes listed.
E.	BamH1
F.	HindIII, AluI
G.	EcoR1
H.	SmaI
I.	SmaI and HbaI
J.	HbaI
K.	HindIII
L.	AluI, HbaI
M.	EcoRI, AluI
N.	AluI
O.	BamHI, AluI

sangriapug213

DNA is isolated from person X and is seperately digested with restriction enzymes EcoRI and Hhal. What is the expected occurrence of restriction sites for Hhal? Which enzyme is expected to produce the lower % of smaller restriction fragments? Which enzyme is expected to have restriction sites at the lower frequency in the human genome?

taupewater-buffalo614

1. A restriction enzyme has a 6 base recognition sequence. How often would it cut DNA in a typical genome?

2. A linear DNA fragment is cut with EcoRI and HindIII:

EcoRI: 1,5,6 kb fragments

BamHI: 3,9 kb fragments

EcoRI and BamHI together: 1,2,4,5 kb fragments

What is the arrangement of the restriction enzyme sites (E=EcoRI, B=BamHI, numbers indicate distance in kb between restriction enzyme sites)?

Biology 1202B Lecture Notes - Dominance (Genetics), Galactose, Mutagen

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