D DNA REPLIC CATION A ADDITIONAL L PRACTICE E Q Question 1 N05P2Q3 N Fig. 1.1 is a diagram d show wing DNA re eplication.

(a) )

Name two moleculles required for DNA rep plication that are e not shown n in Fig. 1.1. [2]

(d)A ATP, (d)GTP P, (d)CTP , (d)TTP/ eoxyribonucl eoside triphosphatesDN NA nucleotides/ de de eoxyribonucl eotides NA ligase DN prim mase He licase Sin ngle-strand b binding prote eins Top poisomerase e (b) ) Explain n how Fig. . 1.1 show ws semi-con nservative replica ation. [2]

tal DNA mole ecule unzips and separ rates, Parent ach strand o of parental D DNA molecul le is used as sa ea tem mplate to sy ynthesize a complemen ntary DNA s strand the dau ughter/new D DNA molec cule will cons sist of on ne parental/o orginal stran nd and one n newly synthe esized/ da aughter stran nd

(c)

Describe how the two o original st trands deter rmine the structure of the two ne ewly synthes sized strand ds. Complem mentary bas se paring A pairs with w T and C pairs with G

[2] (d d) The DNA D molecule is describ bed as havin ng anti-para llel strands. Describe th he evidence e visible in Fig. 1.1 and Fig 1.2 that DNA has anti-parallel strand s. [2] From Fig 1,2, one e DNA strand d runs in 5 to t 3 directio n, the other strand runs in 3 to 5 di irection DNA polymerase e adds nucle eotides to free f 3OH e ends only, h hence on e each parenta al strand, synthe esis of new strand s is in opposite o dire ection ( from m Fig.1.1)

An nderson Junior CollegeBiology Department 2013

Ev valuation: DNA REPLICATION R

CHALLENGING QUESTION Question 1 CJC 2009 Fig 1.1 shows a section of DNA undergoing replication.

StrandA

StrandB

Fig 1.1

(a)

Which strand of the DNA molecule is being used as a template? Give reasons for your answer. [3] Tempate: Strand B reason: Presence of only deoxyribonucleotides in Strand B indicates parent template reason: Presence of ribonucleotides in between deoxyribonucleotides in Strand A indicates Okazaki fragment

(b)

RNA primer will be replaced by DNA nucleotides by DNA polymerase. Indicate with two arrows on Fig 1.1 where the phosphodiester bonds will be broken by DNA polymerase. [1]

(c)

Give 2 differences in the action of DNA polymerase and RNA polymerase. [2] RNA polymerase adds ribonucleotides DNA polymerase adds deoxyribonucleotides

Anderson Junior CollegeBiology Department 2013

Evaluation: DNA REPLICATION

OR RNA polymerase can initiate synthesis of a polynucleotide DNA polymerase can only add nucleotides to the end of a pre- existing chain that is already base-paired with the template strand.

OR DNA polymerase has proof reading / has nuclease activity to remove mismatched nucleotides RNA polymerase has no proof reading activity/ no nuclease activity

Anderson Junior CollegeBiology Department 2013

Evaluation: DNA REPLICATION

AN NDERSON JUNIOR C COLLEGE H2 BIO OLOGY 96 648 JC 1 NSWERS NAME: AN CL LASS:

Evaluation n: DNA REPLICATION N Q1 Q2 TOTA AL MARKS

16 Question 1 d Fig. 1.1b are a diagrams s showing DNA replicatio on. Fig 1.1a and

Fig. 1.1 (a)(i) [4] With h reference to Fig. 1.1a, describe how nucleotide es are added d to the grow wing DNA st trand.

As A each deo oxyribonucleoside trip phosphate iis added, hy ydrolysis of f pyrophosp phate / 2 phosphate p groups g relea ases energy y to drive the e addition of f nucleotide to the growing DNA strand s phosphodie p ester bonds s formed be etween the 3-OH grou up of the las st nucleotide of the elongating e chain and the e 5-phosph hate group o of the newly added nucle eotide catalyzed c by y DNA polym merase Order O of bas ses added is s determined d through co omplement tary base-pa aring via hy ydrogen bond b format tion with par rent strand which w act as a template [any 4]

Anderson Junior College C Biology Dep partment 2013

Evaluation: DNA A REPLICATION

(ii) With reference to Fig. 1.1b, explain why the two elongating strands are synthesized in opposite directions. [3] Meaning of opposite direction ( from Fig) Leading strand is synthesized towards the replication fork while lagging strand is synthesized away from the replication fork. Reason DNA polymerase can only add deoxyribonucleotides to a free 3 end The parental/ template DNA strands are anti-parallel to each other

(b)

Explain why RNA polymerase is required in DNA replication. [2] RNA polymerase synthesizes a primer which provides a base-paired free 3 end for DNA polymerase to add deoxyribonucleotides to [Total: 9]

Question 2 NYJC2007 / SAJC2006 Promos The cell cycle comprises of two main stages, namely interphase and mitosis. During interphase, DNA replication occurs at synthesis phase, before mitosis can take place. In an investigation of DNA replication, a species of bacterium was grown for many generations in a medium containing 15N isotope of nitrogen. The culture was then switched to a medium containing only 14N for one generation of growth; it was then returned to a 15N-containing medium for two more generations of growth. DNA was then isolated from the bacterial cells and the density of the DNA was measured using density gradient centrifugation. In this technique, samples of different densities settle at different levels in the tube. Tube A and B of Fig 2.1 below shows a key to represent the positions of 15N- and 14N-labelled DNA respectively.

Anderson Junior CollegeBiology Department 2013

Evaluation: DNA REPLICATION

(a)

Describe the semi-conservative mode of DNA replication. [3] The parent molecule unzips and separate, each strand act as a template to synthesize the complementary DNA strand Each newly synthesized DNA molecule consists of one parent/ original strand and one daughter/new strand In the Tube C draw and label the possible band (s) that can be obtained at the end of two generations of growth in 15N-containing medium. [2] 2 DNA bands, one at the 15N position, one at position halfway between that of 15N and 14 N Thickness of 15N-15N DNA band is 3 times that of 14N-15N DNA band At the end of two generations of growth in 15N-containing medium, state, (i) the percentage of 14N containing bacteria cells obtained [1] 25% (ii) the ratio of 14N-labelled to 15N-labelled DNA strands obtained [1] 1: 7 [Total: 7]

(b)

(c)

Anderson Junior CollegeBiology Department 2013

Evaluation: DNA REPLICATION