Mapping in C.

elegans to assign chromosomal linkage of spe

gene.
Alyce, Jashayla, Keonna, Shifat

Abstract
C. elegans are small roundworms that are very useful for research. They have many

essential characteristics that helps the central and biological problems of the human body. These

worms have six chromosomes which consist of five autosomes and one sex chromosome that

determines the sex. The sex of the worms will be hermaphrodites or males. In this experiment,

we are researching whether the spe gene is on the same or a different chromosome.

Methods that were used to see if the chromosome were on the same or a different

chromosome were by cross mapping the gene to the assigned chromosomes. The chromosome

with the lowest number of spe wells contains the spe gene. Chromosome IV had low spe wells

but it was not low enough to consider the spe gene is on chromosome IV. If the spe gene is on

the chromosome, we expect none or much less than ¼ wells to be spe. If spe is not on the

chromosome we expect ¼. In the results, it is concluded that the spe gene is on chromosome II

because it has less than expected spe wells and had closer to ¼ spe wells.

Introduction
Caenorhabditis elegans are non-pathogenic, non-hazardous-free living transparent

nematode worms. These roundworms survive feeding on microbes, primarily bacteria, and live

in the soil, more so rotting vegetation. Caenorhabditis elegans share many essential biological

characteristics that are central problems to human life. These worms are conceived as a single

cell that undergoes the development of embryonic cleavage while proceeding to grow as an

adult. There are two sexes: a self-fertilizing hermaphrodite and a male that produces sperm, egg,
mating and reproduces. These worms have been used in research studies on the genetic

regulation of aging. Also, are used in understanding the cellular mechanisms of learning and

memory because of their small nervous system.

C. elegans uses genetic nomenclature for the naming of genes, mutations, strains, etc. In

this experiment over five consecutive weeks, we performed genetic crosses that mapped

spermatogenesis defective (spe) mutant to other C. elegans chromosomes. However, in C.

elegans if there is a mutation in a spe gene the worms are sterile because something is wrong

with the sperm in fertilizing the oocytes. We are studying how sperm and eggs interact and fuse

to form a zygote during fertilization. However, if the worms are tampered with mutagenic

chemicals, we are looking for worms where the process is broken. We look for proteins that are

required for fertilization.

Material and Methods

The C. elegans experiment consisted of five dedicated weeks. The first week, we had 12

L4 or wild types and 3 L4 stage hermaphrodites. As we continue the process the hermaphrodites

will not be together but separate from young larvae and eggs. The L4 mutant hermaphrodites

were being moved to mate. The wild type L4 adults had to wait until the L4 mutants. The worms

were placed to be incubated for three days at 15 degrees Celsius and some of the adults will be

moved to 20 degrees Celsius for four days. The males and hermaphrodites created a genotype. A

mate cross of 12 mutant/+ males into 3 spe/spe hermaphrodites. The cross that was used UNC5

chromosome 1; unc 5 IV/SPE. The first plate will be incubated for three days at 15 degrees
Celsius and the adult worms on plate B will be at 20 degrees Celsius for four days. At this stage

of the process, we picked 12 hermaphrodites so they will be fertile. The first set would be

incubated at 15 degrees Celsius, and adults' worms will be on plate B at 20 degrees Celsius for

four days. The worms from each process have now ¼ mutant worms and ¾ wild types. 24 L4

mutant UNC 5 chromosome 1. This will be incubated at 15 degrees Celsius for a week. The end

of the experiment in most cases, we used the 24 wells plates that is for the worms that are sterile

and fertile after the being in the incubator.

Results and Discussion

After counting the 24-well plates for spe progeny of the entire class, we found that 23 of

82 wells are spe for unc-13 (I) mutation, 5 of 95 wells are spe for rol-6 (II) mutation, 20 of 93

wells are spe for unc-32 (III) mutation, 10 of 70 wells are spe for unc-5 (IV) mutation, 14 of 71

wells are spe for dpy-11 (V) mutation, and 21 of 96 wells are spe for lon-2 (X) mutation (Table

1).

Chromosome spe Wells/Total Wells

Unc-13 I 23/82

Rol-6 II 5/95

Unc-32 III 20/93

Unc-5 IV 10/70

Dpy-11 V 14/71

Ion-2 X 21/96

Table 1: number of wells with spe worms counted in the 24-well plates for each mutation marker
gene.
 If the spe gene was on the same chromosome as the mutant marker gene, the progeny

would have no mutant/spe genotype as the two genes would be linked and there would be no

independent segregation. The unc-5/spe (IV) cross is shown below (Figure 1) as an example:

Unc-5+/spe (IV) Unc-5/spe+ (IV)

Unc-5+/spe (IV) Unc-5+/spe Unc-5/spe+

Unc-5/spe+ Unc-5+/spe

Unc-5/spe+ (IV) Unc-5+/spe Unc-5/spe+

Unc-5/spe+ Unc-5/spe+

Figure 1: A self-fertilization cross of unc5/spe (IV) hermaphrodite showing all of the possible
progeny if the spe gene is on chromosome Iv as the unc-5 gene

If the gene was on chromosome II as the unc-5 gene, as expected from the cross, we

would not see any unc-5/spe genotype in the progeny.

If the spe gene was not on the same chromosome, 1/16 of the progeny will be mutant/spe

due to independent assortment. Since for this experiment, we only used unc worms in the 24-

well plates, the progeny probability will show ¼ of the worms to be unc-5/spe (Figure 2).

Unc-5 Unc-5 (IV)+/spe Unc-5 (IV)/spe+ Unc-5 (IV)/spe

(IV)+/spe+

Unc-5 Unc-5 Unc-5 (IV)+/spe Unc-5 (IV)/spe+ Unc-5 (IV)/spe

(IV)+/spe+ (IV)+/spe+ Unc- Unc-5 Unc-5 Unc-5

5 (IV)+/spe+ (IV)+/spe+ (IV)+/spe+ (IV)+/spe+

 Unc-5 (IV)+/spe Unc-5 Unc-5 (IV)+/spe Unc-5 (IV)/spe+ Unc-5 (IV)/spe

(IV)+/spe+ Unc-5 (IV)+/spe Unc-5 (IV)+/spe Unc-5 (IV)+/spe

Unc-5 (IV)+/spe

Unc-5 (IV)/spe+ Unc-5 Unc-5 (IV)+/spe Unc-5 (IV)/spe+ Unc-5 (IV)/spe

(IV)+/spe+ Unc-5 (IV)/spe+ Unc-5 (IV)/spe+ Unc-5 (IV)/spe+

Unc-5 (IV)/spe+

Unc-5 (IV)/spe Unc-5 Unc-5 (IV)+/spe Unc-5 (IV)/spe+ Unc-5 (IV)/spe

(IV)+/spe+ Unc-5 (IV)/spe Unc-5 (IV)/spe Unc-5 (IV)/spe

Unc-5 (IV)/spe

Figure 2: A self-cross of an unc-5(IV)/spe hermaphrodite showing all of the possible

progeny if the spe gene is not on chromosome IV as the unc-5 gene.
*Shaded cells showing the probability of getting unc worms in the progeny. Dark grey cell
showing the probability of the unc worms also being spe.
As discussed above, when the spe gene is on the same chromosome as the mutant gene,

there will be no mutant/spe progeny. However, we might still expect some progeny with this

genotype because of recombination between the genes. In this case, the mutation, which is on

same chromosome as the spe gene, will show very few mutant worms being spe as well. Out of

all the mutant genes, the lowest number of spe worms in progeny is from rol-6 (II) and unc-5

(IV) is the second lowest. If the spe gene is not on chromosome IV; the progeny would have ¼ or

approximately 17 spe worms. On the other hand, if the spe gene is not on chromosome II, the

expected number of spe worms in the progeny would be 22. Even though unc-5 (IV) mutation

has less spe worms than expected, it can happen due to the total progeny being smaller than the
rest of tests. The rol-6 (II) shows very few spe worms compared to the expected amount in the

case that the spe gene is not on chromosome II. Therefore, the spe gene must be on chromosome

II as the rol-6 gene.

References:
“Caenorhabditis Elegans Genomic Similarities to Humans and Their Importance in the Study of
Gene.” Actforlibraries.org, http://www.actforlibraries.org/caenorhabditis-elegans-
genomic-similarities-to-humans-and-their-importance-in-the-study-of-gene/.

“Caenorhabditis Elegans.” Caenorhabditis Elegans - an Overview | ScienceDirect Topics,

https://www.sciencedirect.com/topics/neuroscience/caenorhabditis-elegans.

“College of Biological Sciences.” What Is C. Elegans? | College of Biological Sciences, 27 Oct.

2021, https://cbs.umn.edu/cgc/what-c-elegans.

Mack, Hildegard I.D., et al. “The Nematode Caenorhabditis Elegans as a Model for Aging
Research.” Drug Discovery Today: Disease Models, Elsevier, 6 Jan. 2019,
https://www.sciencedirect.com/science/article/abs/pii/S1740675718300343#:~:text=The%
20nematode%20Caenorhabditis%20elegans%20is%20a%20key%20model,way%20toward
s%20many%20important%20discoveries%20in%20this%20field.

Z.F, Altun, and Hall D.H. “Introduction to C. Elegans Anatomy.” Hermaphrodite Introduction,
2009, https://www.wormatlas.org/hermaphrodite/introduction/Introframeset.html.