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BIOL 360 Sapling HW 9
Terms in this set (6)
Suppose you have a DNADNA fragment you would like to insert into the pSAPpSAP plasmid. The fragment has PstI and EcoRI restriction endonuclease sites near the 5' end and HindIII and Smal restriction endonuclease sites near the 3' end.
Choose the one or two best restriction endonucleases to digest both the DNADNA fragment and pSAPpSAP
EcoRI & HindIII
Order the steps to generate a transgenic mouse from when male and female mice are first mated to breeding heterozygous offspring together to generate a homozygote.
1. mate male and female mice
2. harvest fertilized eggs from mated females
3. inject the transgene into a pronucleus
4. implant the embryos into a surrogate female
5. identify integrants by DNA analysis
6. breed heterozygous offspring together to generate a homozygote
Electrophoresis gels are used to separate DNA fragments. Match the DNA fragment size, in base pairs (bp), to each band on the electrophoresis gel. A DNA ladder with known fragment sizes is visible in lane 1. The sizes of the most intense bands in the ladder are provided to the left. Electrode charge is indicated to the right of the gel.
In the diagram below, the BRCA1 gene is indicated. BRCA1 stands for Breast Cancer Associated 1 and mutations in this gene increase the incidence of breast cancer. It is found that part of the normal sequence of BRCA1 is GAATTT and a mutant allele of BRCA1 is a T to C mutation. This T to C mutation creates an EcoR1 restriction site in the mutant allele.
A genotyping assay is developed whereby the BRCA1 gene is amplified by PCR and subjected to restriction analysis with EcoR1.
Select the gel image that correctly indicates the pattern expected from PCR products amplified and subjected to restriction analysis with EcoR1 for the three possible genotypes.
homozygous for WT -> one band at 1000 bp
heterozygous -> 1000 bp fragment from WT & mutant would be chopped into two unequal fragments
homozygous for mutant -> two smaller size bands
The following diagram illustrates a transposon insertion in the SBEI gene that causes wrinkled peas. The wildtype allele is specified as W and the mutant allele is specified as w.
A PCR assay is used to determine the genotype of different pea plants that are either WW, Ww or ww. The assay works by performing PCR with primers flanking the SBEI gene. These primers are indicated in the drawing with arrows. The SBEI gene, without the transposon insertion, is about 1000 bp. The transposon insertion is about 1000 bp.
Which of the following gel images indicates the correct results for the three genotypes?
In a random sequence consisting of equal proportions of all four nucleotides, what is the approximate average spacing of restriction cleavage sites for a restriction enzyme that cuts a six base-pair sequence?
approximately every 4,000 base pairs
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