BCOR11 Laboratory Exercise 9: Mendelian Genetics NOTE: The main purpose of the exercises in this lab is to give you

practice applying your understanding of Mendelian genetics to determine the patterns of inheritance and linkage of various Drosophila melanogaster genes. You should revie !hapter "# pages $%$&$'( and !hapter "( pages $)$&$)' in Campbell Biology, 9th Edition by *eece+ et. al. ,E-O*E you come to lab. ,e sure you understand ho to construct .unnett s/uares and ho testcrosses can be used to determine hether specific alleles are dominant or recessive or somatic or sex&linked

Outline of Activities and Objectives 0. 1nalysis of ,acterial Transformation .lates 2 rap up of 3abs ' and 45 &1ssess bacterial colony formation on the 3, agar plates prepared in 3ab 4 &1ppropriately document your results and observations 00. Observation of live Drosophila melanogaster &6ain experience identifying phenotypes and sexes of Drosophila melanogaster 2fruit flies5 6etting to 7no 8irtual -ly3ab &1ppreciate the usefulness of laboratory simulations &9nderstand the purpose and limitations of the 8irtual -ly3ab program &.ractice performing and analy:ing crosses using 8irtual -ly3ab Drosophila melanogaster 6enetics: .atterns of 0nheritance &!onstruct .unnett s/uares and generate predictions that ill allo you to assess the pattern of inheritance for a single trait &.erform crosses to determine the patterns of inheritance for various Drosophila melanogaster traits

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08.

,efore scientists had even observed chromosomes or kne that ;N1 as the genetic material+ 6regor Mendel<s analyses of the patterns of inheritance for various traits in pea plants provided a set of rules that formed the basis of modern genetics. These rules of Mendelian genetics are at the core of all branches of biology. 7no ledge of these rules allo s biologists to predict the patterns of inheritance of many traits in virtually all organisms. 1s you discussed in lecture+ some human traits follo Mendelian patterns of inheritance thus allo ing doctors to predict the probability of inheriting certain genetic disorders. Mendel<s rules also help evolutionary biologists to understand ho traits evolve. 1lthough+ as you kno + not all traits follo simple patterns of Mendelian inheritance+ a solid understanding of Mendel<s rules is re/uired to understand more complex patterns and essential for biologists in all fields. The goal of this lab is to further develop your ability to apply your kno ledge of Mendelian genetics to assess patterns of inheritance and gene linkage. Analysis of Bacterial ransfor!ation "lates
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0n 3abs ' and 4+ you isolated a plasmid containing a gene that confers ampicillin resistance+ visuali:ed your plasmid+ and performed a transformation to =insert> the plasmid into a strain of E. coli that lacked an ampicillin resistance gene. You then set up four 3, agar plates ith bacteria+ ? plates ith ampicillin+ " lacking ampicillin to allo you to determine hether or not your transformation as successful. No is your opportunity to observe the final results of the experiment and make your o n @udgments about ho ell your transformation orked. 1s you look at your plates+ you should think about the follo ing /uestions: "5 Ahich plates have no coloniesB Ahich ones are covered ith bacteria 2called a =la n>5B $5 -or plates that have individual colonies that can be distinguished from one another+ ho many are thereB ?5 ;escribe hat the positive and negative controls are: #5 Ahat can each plate tell you about ho successful your transformation asB Make sure that you carefully document your results 2including the number of colonies you counted5 because you ill need this information to rite your *esults section for the experiment.

Observation of Live Drosophila melanogaster Drosophila melanogaster 2fruit files5 are an ideal model organism for studying patterns of inheritance because their genetic information is contained ithin only four pairs of chromosomes. 0n addition+ they reproduce relatively /uickly and generate hundreds of offspring each time they mate. -or this portion of the lab+ you ill observe ild&type flies as ell as flies ith several different mutations and learn ho to distinguish bet een males and females. .henotype 1nalysis There are three types of flies for you to observe: ild&type+ vestigial inged+ and hite&eyed. !arefully examine each type of fly and fill in the table on the attached assignment describing their characteristics. Cex ;etermination To assess hether or not a gene is sex&linked+ it is very important to be able to distinguish bet een male and female flies. There are t o characteristics that can be used. The rear ends of females are more pointed 2=tulip& shaped>5+ hile males< backsides are more rounded and darker. ,ecause shape is a /ualitative characteristic+ male flies are definitively distinguished from female flies by the presence of sex combs on their front legs. Cex combs are a fringe of black bristles on the front legs of male flies that look like black bracelets. Examine each of the three types of flies. 3ook at ho the phenotypes differ. 1lso practice distinguishing male and female flies.

Gettin# to $no% &irtual 'lyLab No that you have gained experience examining phenotypes and determining the sexes of Drosophila melanogaster+ you ill perform experiments to analy:e the patterns of inheritance for various traits using 8irtual -ly3ab. 1lthough Drosophila melanogaster reproduce rapidly+ it can take months to perform enough crosses to properly determine the pattern of inheritance for an individual trait. To allo you to obtain results from today<s experiments much more /uickly+ you ill use 8irtual -ly 3ab+ a computer simulation that ill allo you to get hat all researchers ould love D instant results. 9sing the simulation+ you can accomplish lots of crosses ith simulated groups of "EEE flies in a very short time. 1lthough the experiments you<ll be performing today using the 8irtual -ly3ab program ould certainly be feasible to do in =reality+> there are many situations here computer simulations are crucial because they allo researchers to perform experiments that ould take years or are simply not possible. -or these introductory exercises and the experiment you ill perform in the follo ing t o sections+ you ill ork ith a partner. ,e sure that each of you gets a chance to set up crosses and that you both record all of your results and observations on your assignments. You ill be performing experiments in this lab to assess the patterns of inheritance of various fruit fly traits using 8irtual -ly3ab. 0t is important that you understand ho this program orks and its limitations so that you can properly design your experiments and analy:e your results. There are a fe key =soft are rules> for 8irtual -ly3ab listed belo . Many of these rules simplify the analyses+ so they are orth reading carefullyF NOTE: Some of these rules represent constraints that are not present in nature. *efer back to these rules as you perform your experiments. 0n addition+ although some of the mutations in 8irtual -ly3ab are lethal+ the set of mutations you ill be analy:ing does NOT include lethal mutations. The 8irtual -ly3ab rules for lethal mutations are more complicated+ and therefore analy:ing and interpreting lethal mutations can be difficult analy:e using this program. -ly3ab =*ules> & 1ll mutations sho complete dominance 2 hat does this meanB5 & Mutations are organi:ed into different groups based on their phenotypes & 0f one of the parents carries mutant alleles for the trait+ the other must be ild&type & Only one mutation from each group can be selected for an individual fly & The default setting is ild&type+ so unless you choose a mutation+ the fly ill have t o ild&type alleles & 0f a mutation is selected+ the fly ill be homo:ygous for the mutation 2Aould this ork for a lethal mutationB5 & 1ll sex&linked traits are located on the G chromosome .ractice To practice using 8irtual -ly3ab+ follo the steps belo to perform a cross bet een a ild&type female and a hite&eyed male and cross the -" generation to obtain an -$ generation. Then do the =reciprocal cross>& cross a hite&eyed female and a ild type male. 6et the analysis of the -" generation and then the -$ generation. TO .E*-O*M THE C0M931T0ONC 1,O8E -O33OA THE -O33OA0N6 ;0*E!T0ONC: ". 6o to biologylabsonline.com. !lick on -lylab. You ill be asked for username and pass ord. This ill be denoted on your computer
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"5 On the =introduction> page of 8irtual -ly3ab+ click on the Ctart 3ab button and ait for the lab to load in a pop&up screen. $5 9nder Each Mating the number of offspring ill be "EEE.. ?5 !lick on the ;esign button underneath the female fly. Mutations are organi:ed into groups listed along the left side. You can see that there are five different bristle mutations. ,ecause you ant the female fly to be ild&type+ simply click on Celect at the bottom 2the default for all mutations is ild&type5. #5 !lick on the ;esign button underneath the male fly. To create a hite&eyed male+ click on the Eye !olor group on the left and choose hite eyes. !lick Celect. (5 No click on the Mate button to perform your cross. The offspring phenotypes appear at the bottom of the screen. 0f you click on the =$ fly images> arro @ust above them+ you ill see the parental flies. 0f there ere more than t o offspring phenotypes+ a second arro ould appear belo the offspring+ and you could click on the arro to see the other phenotypes. %5 The easiest ay to see the results of your cross is to click on 1naly:e *esults. This ill sho you the phenotypes separated by sex+ the number of offspring+ the proportions+ and the ratio. 2Ahy is it important the results are separated by sexB5 -ill in the table on your assignment ith your results. 0f there are no flies of a certain phenotype @ust rite :ero. *eturn to the results screen by clicking *eturn to 3ab 2upper left5. '5 Next+ cross the -" generation to produce -$ offspring by clicking the Celect buttons underneath the female and male -" offspring. Once you have selected the flies+ click Mate. Notice that in the -$ generation there are three different phenotypes. !lick on the " more fly arro to vie the third. 45 !lick 1naly:e *esults and record your data on your assignment. Ahat does this tell you about the mutation for hite eyesB )5 Ahen finished+ close the popup indo and select Yes hen a box pops up asking if you really ant to close the indo . No perform the reciprocal cross 2 hite eyed female x ild type male5 suggested above. Drosophila melanogaster Genetics: "atterns of (n)eritance No that you are familiar ith ho 8irtual -ly3ab orks+ you are ready to perform further analyses. 0n the first set of experiments belo + you ill analy:e the -" generations of various crosses to determine hether certain fruit fly traits are "5 dominant or recessive+ and $5 sex&linked or autosomal 2not sex&linked5. Hypotheses I .redictions

To determine the pattern of inheritance of a specific trait+ you first need to generate a hypothesis and then make predictions that allo you to test your hypothesis. ,ecause you need to determine both hether or not a mutation is dominant or recessive and hether it is G&linked or not+ there are four potential hypotheses: "5 ;ominant and autosomal $5 *ecessive and autosomal ?5 ;ominant and G&linked #5 *ecessive and G&linked -or all of your analyses+ you ill perform t o crosses: ild&type female x mutant male and mutant male x ild& type female. Construct the necessary Punnett squares in the space provided on your assignment and fill in the table of predictions for the hypotheses on your assignment. ,e sure that you take into account the rules of 8irtual -ly3ab. Hint D you really only need to dra three s/uares. Experiment Having developed your predictions+ you are no ready to analy:e the patterns of inheritance of several Drosophila melanogaster traits. .erform the crosses indicated in the table labeled =.atterns of 0nheritance 1nalyses> on your assignment. !lick Ne Mate to start a ne cross each time. -or the last cross+ choose a mutation from the set listed belo the table. ecord the phenotypes of the !" generation and determine #hether the trait is dominant or recessive and #hether it is $%lin&ed or not. 9se your predictions to help you figure out the patterns of inheritance. 21lso see =Hints for Your 1nalyses> at the end of the lab handout.5

BCOR11 Lab 9 Mendelian Genetics Assi#n!ent *ue at t)e be#innin# of Lab 1+ Most of your assignment should be completed during your lab period using 8irtual -ly3ab. You ill need to use information your gather from your 8irtual -ly3ab experiments to ans er some of the post&lab /uestions at the end of this assignment. .lease rite your ans ers neatly in the spaces provided. Your completed assignment should be turned in to your laboratory '( at the beginning of )ab "*. NOTE: remember that the outlines for your ,acterial Transformation *esults and ;iscussion are also due at the beginning of 3ab "E. Observation of 3ive Drosophila melanogaster .henotype 1nalysis !ly 'ype Aild&Type 8estigial Ainged Ahite&Eyed Eye Color +ing Si,e - Shape

6etting to 7no 8irtual -ly3ab -" 6eneration Se. -emale -emale Male Male Phenotype ild&type hite eyes ild&type hite eyes /umber Proportion atio

-$ 6eneration Se. -emale -emale Male Male Phenotype ild&type hite eyes ild&type hite eyes /umber Proportion atio

Drosophila melanogaster 6enetics: .atterns of 0nheritance 9se the space belo to dra your .unnett s/uares.

-" 6eneration .redictions 0ypothesis ;ominant+ 1utosomal Cross AT female x mutant male mutant female x AT male AT female x mutant male *ecessive+ 1utosomal mutant female x AT male AT female x mutant male ;ominant+ G&linked mutant female x AT male AT female x mutant male *ecessive+ G&linked mutant female x AT male Phenotype !emales Phenotype 1ales

.atterns of 0nheritance 1nalyses !ross .arental -emale ild&type curved ings ild&type yello body lobe eyes ild&type
G G G G G G G

.arental Male curved ings ild&type yello body ild&type ild&type lobe eyes ild&type

.henotypes -" -emales -" Males

.attern of 0nheritance ;ominantJ*ecessive G&linkedB

"

ild&type

!hoose a mutation from the follo ing list for the fourth cross: miniature ings+ crossveinless ings+ shaven bristles+ singed bristles+ bro n eyes.

"ost,Lab -uestions "5 You had the opportunity to observe live hite&eyed Drosophila melanogaster as ell as perform crosses ith hite&eyed flies in =6etting to 7no the 8irtual -ly3ab.> a. ,ased on the results of your crosses+ is the hite eye mutation dominant or recessiveB

b. 0s the hite eyed mutation sex&linkedB

$5 -or each cross you performed in the =.atterns of 0nheritance 1nalyses+> briefly explain hy you said that the mutation as dominantJrecessive and G&linkedJnot G&linked. !urved Aings

Yello ,ody

3obe Eyes

KKKKKKKKKKKKKKKKKKKK 2mutation of your choice5

Also due at t)e be#innin# of lab 1+ is your Bacterial ransfor!ation (ntroduction. "lease refer to your /A student )andboo0 for %ritin# in biolo#y1.

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