Concept explainers
Sickle cell anaemia is caused by a mutation in the HBB gene. The normal wild-type βA allele contains an MstII restriction site; in the mutated sickle-cell βS allele this restriction site has been lost.
PCR amplification of part of the gene encompassing the mutation site from a normal unaffected individual results in a product of 110bp. Digestion of the PCR product with MstII and subsequent gel electrophoresis results in a single band of 55bp.
How many bands and of what size would you expect to see in individuals who:
i) has sickle-cell anaemia (has two βS alleles)
ii) carry the disease (sickle cell trait) (has one βS and one βA allele)
a. |
|
||||||||||
b. |
|
||||||||||
c. |
|
||||||||||
d. |
|
Step by stepSolved in 2 steps
- Based on the restriction enzyme specificities given below, what was the enzyme utilized to produce the overhangs in the cut DNA sample? Restriction Enzyme Specificities: ... GAATTC... Hpal ....СТТААG... GTTAAC... .CAATTG... PstI ..CTGCA.. ..GĄCGTC.. EcoRI Hind III ... AAGCTT... Bam HI ...GGATCC... ..ССТAGG. .TTCGAA... DNA sample: GATCC- ICCTAG ССТAG A) BamHI в) Нраl ЕcoRI (D) Pstl E) Hindlllarrow_forward(b) The treatment given to the patient was 10 mg of menadione (formula on the right) every 6 hr with a dose of 1 g of ascorbate (Vitamin C) every 6 hr. The rationale for this treatment was that menadione (also called Vitamin K3) could accept electrons from reduced CoQ of the electron transport chain, shuttle them to reduce ascorbate, which, in turn, could reduce cytochrome c(Fe3*) to cytochrome c(Fe2*) that in turn becomes the substrate of Complex IV or cytochrome c oxidase. This artificial sys- tem would bypass the defective Complex III site in the electron transfer chain. To show the effect of this regimen with respect to the ADP phos- phorylative capacity of her skeletal muscle, corresponding 31P NMR spectra are shown on the right. „CH3 2-methyl-1,4-naphthoquinone (menadione)arrow_forwardExperiment 1: An RT-PCR was run on human cells growing in tissue culture under standard conditions, using the primers described above. A normal PCR was also run on the cells, using the same primers. They were run on an agarose gel shown on the left in the figure (gel 1), in the wells indicated by ‘RT-PCR’ and ‘PCR’. The arrow shows the direction of current in the gel. a) Observe the gel and describe the results for the RT-PCR and PCR (no explanation, just describe). b) Explain why the bands have travelled different distances on the gel.arrow_forward
- In biotechnology, gene cloning is a crucial technique used in many genetic modification experiments. A vector is normally required to perform this process. The vector commonly used to transform a bacterial host cell is known as a plasmid. Below is the general plasmid map of pBR322. Elaborate the functions of ori, Ap and TeR present in pBR322. Bul 3759 Prul 3733 PM 3507 BDI 3602 Asel 3537 Bal 3433 Finci 3905 Scal 3844 3787 Bad 3420 Ahdi 3361 Acul 3000 AlwNI 2884 Sspl 4168 Earl 4155 Acul 4048 Xust 3961 Bell 2777 Bc1 2682 ori Aall-Zeal 4284 BeiVI 4209 B 4205 Del 2575 Pail-Att 2473 BAB 2404 EcoRI 4350 Earl 2351 BspQI-Sapl 2350 Ndel 2295 BAP 2291 178-Accl 2244 Cial-Ispit 23 Hindill 29 pBR322 4,361 bp Ball 2225 TthJ11- Part 2217 rop EcoRY 185 Bmtl-Nhel 229 Hamill 375 Sgr 400 Ball 471 Xml 2029 Pull 2064 Dalt 2162 BamB 2122 Banil 485 Bigl 128 Sphl 582 EcoN1 822 Sall-Accl-Hell 651 Pshu 712 BsaBl 1668 Engl 939 BeY1 945 Nrul 972 BAPT 1045 BspMI-BfuM 1063 PAMI 1315 Bumi 1353 Bgl 1650 BapEl 1664…arrow_forwardThe PET28 plasmid is cut with the Xhol and Ncol restriction enzymes. A 200 bp gene, also cut with Xhol and Ncol, is ligated to the large segment of the plasmid. In the new plasmid, what is the location of the Xbal restriction site? O 335 bp O 535 bp 397 bp O 197 bparrow_forwardAnswer the question in the photoarrow_forward
- Recently, I am designing primers specific for Lactobacillus sp. (partial sequence of the 16s rRNA gene) and can get successful amplification on my conventional PCR. Then I am going to quantify by RT-PCR using SyBr Green Reagent in Step One Plus Instrument. By using the same run condition as the conventional PCR, I face the problem that many of my samples got a bad melting curve. Anyone have the same experience as me or suggestion please let me know. Thanks in advance.arrow_forwardIn the following "gene library" cloning experiment Digested genomic DNA AmpR gene TCR gene TCR is tetracycline resistant marker, AmpR is ampicillin resistant marker and BamHI is the unique restriction enzyme on plasmid. A PhD student digests/cuts the plasmids with BamHI restriction enzyme and the genomic DNA with EcoRI restriction enzyme. After performing the cloning experiment and obtaining colonies on a selection plate, the obtained cells will be ..... (Hint: this question is even more challenging; the PhD student was later demoted to an MSc student). a) resistant to ampicillin and tetracycline b) sensitive to tetracycline and ampicillin c) resistant to tetracycline and sensitive to ampicillin d) resistant to ampicillin and sensitive to tetracycline e) sensitive to ampicillin and tetracycline BamHIarrow_forwardBelow are several problems frequently faced by researchers when running the PCR. Give one (1) solution to each problem stated. 1. No PCR product 2. Multiple bands appeared after gel electrophoresis (you are amplifying just 1 gene) 3. Bright thick bands at the end of the agarose gel after electrophoresis 4. Bands appeared at the negative controlarrow_forward
- The gel below shows results for the bitter tasting SNP analysis from a CH306 class from a few years ago. Analyze the results and annotate the gel to indicate the bitter tasting ability and homozygous/heterozygous status of each of the individuals represented on the gel. There are three lanes of markers, and the first non-maker lane on the left of the gel is an uncut control PCR product (i.e. there are a total of 10 individuals on the gel starting in lane 3).arrow_forwardTitled “Techniques utilized to generate a genetically engineered organism and confirm gene disruption.” a. In the table, indicate with an "x" if the reagent or technique applies to DNA, RNA, and/or protein. You can have more than one "x" in each row or none at all.arrow_forward
- BiochemistryBiochemistryISBN:9781319114671Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.Publisher:W. H. FreemanLehninger Principles of BiochemistryBiochemistryISBN:9781464126116Author:David L. Nelson, Michael M. CoxPublisher:W. H. FreemanFundamentals of Biochemistry: Life at the Molecul...BiochemistryISBN:9781118918401Author:Donald Voet, Judith G. Voet, Charlotte W. PrattPublisher:WILEY
- BiochemistryBiochemistryISBN:9781305961135Author:Mary K. Campbell, Shawn O. Farrell, Owen M. McDougalPublisher:Cengage LearningBiochemistryBiochemistryISBN:9781305577206Author:Reginald H. Garrett, Charles M. GrishamPublisher:Cengage LearningFundamentals of General, Organic, and Biological ...BiochemistryISBN:9780134015187Author:John E. McMurry, David S. Ballantine, Carl A. Hoeger, Virginia E. PetersonPublisher:PEARSON