During plasmid extraction, after the addition of acetate-conta in ing neutralization buffer the large. plasmids can renature and stay in solution. DNA and prote ins precipitate, but the small bacterial DNA O Autosome O Plasmid O Gen omic O Mitotic O Karyotype
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- Kha Vu Danels Include: 8DX : Safehy Jor bromie tnto lab repart! Name Section date sheet MAPPING PRACTICE #1 Below is a restriction map for the plasmid PGEN 101 (total length = 20 Kb). Using this map as a guide, give the number of restriction fraqments along with their associated lengths that would result from digesting PGEN 101 with the restriction enzymes EcoRI, BamHI and a combination of ECORI and BamHI. BamHI 3.2 Kb 1.7 Kb EcoRI BamHI PGEN 101 8.7 Kb 5.5 Kb .9 Kb EcoRI ECORI DIGESTION PERFORMED SIZES OF FRAGMENTS OBTAINED 10.4 kb , 0.9kb, 8.7 Kb EcoRI 3.2 Kb, 16. 8kb BamHI EcoRI + BamHIA amp PBR322 4301 fot B Clear Zones Figure 2 The postgraduate student, Demika, inserted her gene of interest into the plasmid, pBR322, before transformation into the competent host cell using heat shock method. After that she cultured the cells on the Ampicillin agar plate before replica plating the colonies onto another Ampicillin (A) and Tetracycline (B) agar plates shown in Figure 2. (1) Referring to the vector pBR322 in Figure 2, which recognition site was cleaved to insert the gene of interest? Based on the observation above, can you identify which colonies are carrying positive mcombinants of BR322? Explain your selection.What type of molecule or cell is PGLO? C Origin region Arabinose operon arac ori Ndel Promoter PaaD PGLO (5,400bp) Ndel Ampicillin Resistance Gene Amp Hindll Ndel Green Fluorescent Protein (GEP) Gene Hindll 'ECORI GraphiceESchmid/2003 D O virus O RNA plasmid molecule O DNA plasmid moelcule O none of the choices are correct
- Identify types of plasmid( nicked, linear, supercolied and circular,stranded on each bandFIGURE 2 shows the only suitable DNA restriction site in a plasmid DNA vector that can be cleaved. 5'.GTAATCCGGATCCGAAATCCCCCGG... 3' 3.CATTAGGCCTAGGCTTTAGGGGGCCC... 5 FIGURE 2 Identify the most suitable enzymeto be used as a restriction enzyme forthis purpose. O Ligase O Smal O EcoRI О ЕсoRVArial BIUA 11 + .. | I 1 I 3 I 4 i.) Fill in the table for each of the E. coli: (0) = No Activity (+) = Basal Activity and (+++) = High Activity E. coli chromosome F' Plasmid B-gal activity? Permease activity? When When When When Glucose is Lactose is Glucose is Lactose is present present present present a.) I+ P+ O+ Z+ Y+ Inone +++ +++ b.) I^[S] P+ O+ Z+ Y+ none c.) I+ P+ O^[c] Z+ Y+ none d.) I+ P+ O- Z- Y+ none e.) I+ P+ O+ Z+ Y+ I^[S] P+ O+ Z+ Y+ f.) I^[S] P+ O+ Z- Y+ I+ P+ O^[c] Z+ Y- g.) I^[TB] P+ O+ Z+ Y 1+ P+ O^[c] Z- Y+ h.) I+ P+ O^[c] Z+ Y- I+ P+ O+ Z** Y+ i.) I^[TB] P+ O^[c] Z+ Y- 1+ P+ O+ Z- Y+ Z** is a polar mutation ii. ) If the lac operon in 'a' carried a mutation in the CAP binding site that rendered it nonfunctional, how would that affect the level of ß-galactosidase protein activity with and without lactose present, why? MacBook Air 000
- d. Biuret test O e. NaOH Clear my choice In the method to isolate the DNA from peas, you used the centrifuge right after the addition of salts and alcohol; in which part of the tube will you find the stringy white DNA? of Select one: stion a. Suspended in the solution O b. At this step, no DNA will be visible O c. Top of the tube precipitate O d. Bottom of the tube precipitate Clear my choiceConjugation using ________ will result in the transmission of a segment of chromosomal DNAfrom one host to another using a plasmid. Question 25 options: F‑strain High frequency recombination (hfr) strain F+strain F plasmid F' plasmidTo identify mutant genes, a wild-type bacterial genomiclibrary in plasmids is used to transform the mutant bacterialstrain. A transformed ________ in which the mutantphenotype is rescued to wild type likely harbors a plasmidcontaining a wild-type copy of the corresponding gene
