test1 - BlOL 230 Genetics Test 1 name When you finish...

Info icon This preview shows pages 1–6. Sign up to view the full content.

Image of page 1

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

Image of page 2
Image of page 3

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

Image of page 4
Image of page 5

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

Image of page 6
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: BlOL 230 Genetics Test 1 name: ' When you finish, please sign the pledge (but only if it is true!): “On my honor. I pledge that l have neither given nor received any aid on this exam. and I will not/did not discuss it, directly or indirectly. with any other student until after 11:00 AM today. I understand that to do so would constitute a violation of the Xavier University policy on academic honesty and could result in a O on this test."- Since tests are worth 60% of the grade, each test will have 60 points. Questions are worth 2 points unless otherwise indicated. Basic Questions 40 points 1. On the following segment of a longer DNA molecule, identify and label (write specific name): tafour different nitrogenous bases (5 pts) su 7 Vfimple of a bond between two nitrogenous bases / the "ends" of this short segment (C7; mac inf- a .< = ”'41 ”if? (:70! r) (elf-av “Ly 2. A fragment of double-stranded DNA is found to have 1000 nucleotide pairs. The base composition is found to be 46% G + C. (3 pts) 5a.}; I." a. How many nitrogenous bases does it contain? Z 00:: ‘9? b. What % of the bases are thymine? Z 7 / 3. What is the significance of ddNTPs in a sequencing reaction? A. They lack a 3' OH group. 0 9 if B. They terminate the elongating DNA strand. 0=§-f-r-0 ° BASE C. They carry a fluorescent tag and thus mark the last base in a chain. 0 o o D\. Despite being artificial, they are readily used by DNA polymerase. n H @all of the above 4. Imagine that you are very small and you are scuba diving in a tube containing an in vitro DNA replication using the normal enzymes prepared from E. coli. Choose two of the following enzymes/proteins and for each. describe what you would observe if it were accidentally left out of the reaction. (4 pts) Ligase helicase primase single-strand binding proteins 1’ 3A3)“ Q‘OMZMM Wavmu" Q-"k "3;". gender: " 8 U r ’1 . y. c L. It t-LL wen/570“ 7" :rnli" [bi/Vs- TWG; (7.4.:- -:;‘. z— ”I, 9 .} (PROSPEAO'O; CQNL'BONDS l// ' 0 NW I” C’gd W I“ W \ ““053 'fi’rJWK Luck. 0% .7 U-eifcagL ['le . x. 31:1. RUG-UL ST“. ,__ _ 9 ' ,. f"‘-‘.}l4’:.) ‘ NP“ 1"“ JOLUVU'J‘ WOUL-f) NQ' be ‘ ' V UN . MN? I .- .. ”— New l rc 51pm. ad}; on: / 5. List three differences between DNA replication in vivo and PCR (DNA replication in vitro). (3 pts) ONA 73w. - ., U _, 1/- US l: S out A (PO‘V met/2.455 QC {A Q (Paw mJJZthe ‘ US / . U ,_ r‘ - . “'3 We: 7.; m 1” ., ,, 3.3 J M’ WV firs/.571) 9a.}? at“ ;. van-.4; S! KEANDS Si/LA /; V/L-CI‘.\IR.E> Am (minis) 598.13.“, , 7; TA/ / S _ 0 runs (mic, ( MED Sic/L, "7'.ng Why; f‘“9“:4thJKL, qu WM . In DNA replication, the leading and the lagging strands differ in that _________ (2 pts) A. the leading strand is synthesized in the same direction as the movement of the replication fork, and the lagging strand is synthesized in the opposite direction. ' BC the leading strand is synthesized by adding nucleotides to the 3' end of the growing strand, and the lagging strand is synthesized by adding nucleotides to the 5' end. jig”. the lagging strand is synthesized continuously, whereas the leading strand is synthesized in short fragments that ' are ultimately stitched together. D. the leading strand is synthesized at twice the rate of the lagging strand. 7. Which of the following cloning vectors can accept the largest DNA fragments? (2 pts) A. Plasmid B. Bacteriophage A ©(ACS D. Cosmids 8. Sally has a BS in Biology and is now working towards a PhD in Environmental Science. She is specifically interested in bioremediation of heavy metals, common soil contaminants around industrial sites. Sally wants to study metal homeostasis in fungi and decides to do a Genetic Analysis. (4 pta) Put the steps of Genetic Analysis in the proper order (label 1 — 4): / L Clone and sequence the relevant genes. 1 Determine how many genes are affected among the mutants. __’:'-_ Choose an organism that is amenable to genetic manipulations. Sally chooses the fungus Neurosopora. i Collect or generate mutants. Sally makes up a collection of mutants that are abnormally sensitive to arsenic. The mutants are recessive. 2 /~ ' 5/09 615qu My) 9 Sally obtains six mutant strains of Neurospora that are abnormally sensitive Oto arsenic. She mates each mutant strain with each other mutant strain and record the offspring phenotypes The results are summarized below. (“+” means lives on plates containing 10 lug/ml arsenic “ -“ means dies on plates containing 10 pg/ml arsenic ) Strains ------ Mated -+-+-+--I'IIB Draw appropriate conclusions. (4 pts) M, SUDMLéK gem-.3 _ m - PIS?) fr, acting! Con/0’ ('Alw ’ECQr glint so . , my 9(2 L, ow Tru, Spry-L Gun»; I\/\ M‘ 3 V v‘ 6'14”“ 0 PA" 9° N57 Com?!'m :’ baa»: offal/L" SoTyw, a“: 9,, 52,. 3%,,“ gait-8, I\/‘ 7. ,fi . \ "74/": MM': I T} r V a; "in. w. (“MM ”‘3, crime onto (M So if as ow :2 apt. 51M. 10. Outline a general strategy by which Sally should let one her genes (genes involved in heavy metal homeostasis), using the plasmid pNEU. Au“— [Wu (5 pts) 7/ / (A) Gm“. ,. ~» ' V‘ “4"" €~»’ ,t . . -, o I.) .5? NvaOS’Po/LA Lyn-(Lg. cm. CmStrxlig ”25:39.5— / ( JFK/NL/ game To See (,1 r _ / 7 ’ 0H: ‘J‘; N4 / ,/ 'gkmc‘i :VN'JOIVL'Z) I ‘ Z73 «I I . 1'5 In; vegan- \chb (Aw. bk'x‘,“ Plggm'g ’T) NEU / Z) 3(0) , ._,‘ V- ‘1 Sfitfitbg )_ Nun-warp 3 7mg" ’DlWL (”May/163 ’II49.«3¥0.1I-‘n(;l~—_ A a) 901 5AM .m/u-r ._ 777777 __c._ ‘ WW? VféJung 820“ m j 1&5 s- AMER ”Nehru I k -;'QVL,§U‘I¢?. a. Sally could make a genomic library or a cDNA library. To A” pGEM make a cDNA library she would use 5 to make the first 3' 33” strand of cDNA from mRNA. C. an | D' ligase b. g is a restriction enzyme that Sally could use to ”Z reverse transcnptase open up pGEM—Y and to prepare inserts. F. IacZ gene encoding [i-galactosidase R . . . . . ‘6: amp gene encodIng ampICIlhn reSIstance c. To connect the vectors + inserts, Sally will use if . D , l H. cDNA from wild type Neurospora grown in the presence °f arseni" d. To amplify the library. Sally will mix it with E. coli cells. " CDNA from mutant Neurospora grown in the G in the library clones will allow her to select for presence 0f arsenlc transformed cells and kill untransformed cells. 12. Here is a representative of the transformation plates. Assume that millions of E. coli cells were originally spread over the entire plate. The plate was incubated overnight at E. coli’s favorite temperature, 37°C. Explain or generously label the indicated areas of the plate. (3 pts) & 0503 _ H 8mm Cori?” rMpflQSTAA‘p TM )(fi-ml / N - . . uE . ”I .4 V‘le/A— was AW» " ' gawagt f” ‘° 20ng W1 J) ’ 4- Wm "I l " M At; a Z. 311w— ~S 8’2“, Imam" ~ —-7 ”lny ”:92?- «m mac-s mu. M \ vé. {M “ VIM. ) "' 19$an .— rM/1.,x:g TD .. 7h»: - l I A, 10.,“ WAS "“ ' ‘ M . IN» ALL < p. N”- fiNv‘ficw‘i‘lto HAW-’0" ' ._ etween colonies ' a M": Lin“ 5 r M) g’“ . , o . . — Ros.¢1,’rke ' OWL )Nkm WAS Svtfls’r" « Létlo ’— m / ”‘5 ‘ We (o‘md/ white colony ”HM “’ oL/ze/A 1941mm»; ' {M COW-1’03 “3 wL-er, Dip m-P \ L (EH5 Rig: Mewsfimw / M1 ND 013) 055.9 Ramada M N” Qua “173% Z‘IASMK) w: “(“4" {2153' “use 13. Consider the experiments (Introduction and Material & Methods) described on the extra page. (4 pts) p a. Which item ensured that the PCR specifically amplified the desired gene segment? A. 0.25 U DNA polymerase . B. 1x buffer (final MgC/2 concentration 1.5 mM) . 5 ng genomic DNA from cheek cells of 241 unrelated German shepherds. éthe forward (D1c: 5'-CGC GCG TCG GGC CAA GCT G-3’) and the reverse (D2c: 5’-GCG GGG GGC AGG GGG G-3’) primers E. 200 mM of each dNTP b. What was the template for the PCRs? A. 0.25 U DNA polymerase B. 1x buffer (final MgCI2 concentration 1.5 mM) ©5 ng genomic DNA from cheek cells of 241 unrelated German shepherds. D. the forward (D10: 5'-CGC GCG TCG GGC CAA GCT G-3’) and the reverse (02c: 5'-GCG GGG GGC AGG GGG CG-S’) primers E. 200 mM of each dNTP Challenging Questions 15 points 14. Continuation of #8 - 12: Sally has successfully screened her library for a potential hmm gene and recovered one clone. Below is a gel showing the results of several restriction digests of the clone. a. Which digest was planned to give most directly the size of the insert? (2 pts) A. Eco Rl Sal l c. uncut D. Pstl E. an l b. Pay attention to the map of pNEU, and draw a map of Sally's clone. (3 pts) ”WHEN standards II II ECO Rh8'1 2 q 2 Z CJ$ Pstl:7,2 = Oi 2 law’s Sall:6,3 :q : ZcJ's’ anl29 >1 w’ Smmufifiim-l c :-, . _ o I OMS-MO tor/>2 T N O. :L'c 2910/ 57,4: > Z 4? 7 l \ ’ 73 S,\ Q 15. Now Sally wants to find out under what 5 3 conditions the gene is expressed. The gel 2. mm 2. following picture represents a gel and its :5 f, corresponding blot. (3 pts) _. a“ _ n" e e E, E 3 e e E, E 2‘ Samples run on gel: mRNA from Neurospora § F» E g’ g 5 g B E g’ E 5 grown with additives or conditions as E.)- g E 3 a, 3 'g g 9 g E: g indicated. E E g a g g E 3 g a E g Probe: the cloned gene yfg. 9 ”E 'g a ‘- u, E 9 ‘E 'g 8 '- E .r: o L. 1:: c a: A: a) L. n ‘” m .9.) e E m m o N .2» 2 E W m S N .c m u E 3 c ":5 .C to o E 2 C '27; Draw appropriate conclusions. (Bias :5. i: _‘»io{'\'"‘/JC.‘L 13,5» - “N” LION-‘2’“ Q‘ 5‘ “J Evasive .‘o’X - OMQNLL’ 5 Artful. , (. , . w“, mam Wm, Nv-fl‘LLJA;/} AND - to - ,. - Ac 53b M r w ”3‘ ,. Q r: M- irmwu ¢ . .; J , J "* "mt/4 ,7:~< {.J ' ‘ , . 16. You plan a restriction digest of hippopotamus genomic DNA with the enzyme Eae I, which cuts at the recognition site 5'- PyGGCCPu —3’ (Py = pyrimidine. Pu = Purine). Next you want to run a gel to check that digestion was complete. In order to choose the proper concentration of agarose for the gel, you need to know the expected average fragment size. Assume an AT: GC ratio of 50:50. (4 pts) : ' '3 5. / 5 7: . 3' .J w .l \.-. .' \/ ‘-, ",./ .4 . ll ”'27 /’z, ‘i 4‘ ‘v’ ’L ‘ 197;»; ‘M New? ‘EAAérwl-wr 51:2,... 3: N ma: 1;? / 17. One way that cells exert control over a gene’s expression is to modify the DNA itself. A NH; common modification is to methylate cytosines. (3 pts) N / caa‘I— a. Would methylation affect the hydrogen bonding in that region? Why or why not? 0*»! i l 5* .- . $61 .> m; "M t“ -’ 3,. v 0”? 'EW is. —z. P A ,_, ,, . , ‘V V I" ’ M’ ' ’ N ‘ i” "m " N‘ N V1 1 S-Mcl’hflc‘ytosim (M50 W\/\lLV\ 02a; rJO'v' 33:23:": .3'/ Tu“ (if/"3 \/ residue ‘b. it has been observed that methylation inhibits transcription. Briefly speculate how. ”:2 ‘28 L 2' "5:: 'IJ! fjf‘Jx. ,3 7';(3"" an: X "WA.- " ' 5 i 5,. s .. 3. Tag vw claw» Lin-Amy‘s "s ' . . “MN“? #1 . I) LU . O ' - . ’V i"!. 4’5 7r" ;-.:;;7' [o b’: 53:0“ (v mi."— / "In,“ So i "' > I'W4;\£$(.A.:~?i'.'u:\‘ Home U._;:;.'-. C. Emmi. J Remember to sis? honor pledge on the front _ .,._ . ,, 6 V‘ 1 I ’ “We-w» 7306 3..» /O ...
View Full Document

  • Fall '12
  • DorothyB.Engle
  • Genetics

{[ snackBarMessage ]}

What students are saying

  • Left Quote Icon

    As a current student on this bumpy collegiate pathway, I stumbled upon Course Hero, where I can find study resources for nearly all my courses, get online help from tutors 24/7, and even share my old projects, papers, and lecture notes with other students.

    Student Picture

    Kiran Temple University Fox School of Business ‘17, Course Hero Intern

  • Left Quote Icon

    I cannot even describe how much Course Hero helped me this summer. It’s truly become something I can always rely on and help me. In the end, I was not only able to survive summer classes, but I was able to thrive thanks to Course Hero.

    Student Picture

    Dana University of Pennsylvania ‘17, Course Hero Intern

  • Left Quote Icon

    The ability to access any university’s resources through Course Hero proved invaluable in my case. I was behind on Tulane coursework and actually used UCLA’s materials to help me move forward and get everything together on time.

    Student Picture

    Jill Tulane University ‘16, Course Hero Intern