Abstract In this study, we screen for the dumpy phenotype in Caenorhabditis elegans. These worms are shorter and stouter than the wildtype. By screening for the dumpy phenotype among mutagenized worms, we are seeking to find answers how the dumpy First, we address our hypothesis: there are specific genes within the C. elegans genome that code for the biological process of movement and therefore are responsible for the roller phenotype. After isolating mutant roller worms, we can then cross-breed them with wild type worms to test the nature in which the gene is inherited (i.e. dominant or variable penetrance). We can also map the gene associated with producing the roller phenotype, which could then help elucidate the gene homolog in …show more content…
( Darwin J Prockop)
In this experiment we will focus on Ehlers- Danlos Syndrome which is caused by a mutation in the gene COL3A1. Ehlers Danlos syndrome is common name given to more than ten different inherited disorders. All these have a genetic defect in the collagen and connective tissue. Ehler- Danlos syndrome affects the skin , joint and blood vessels. Ehlers- Danlos Syndrome IV is associated with arterial rupture and visceral perforation(Robert A Schwartz,). Currently there is no cure for Ehlers- Danlos Syndrome. What i’m proposing is that through a forward genetic screen you can identify the
The potential to apply forward genetic approaches for comprehensive genetic dissection of vertebrate development was the initial attraction for researchers to utilize the zebrafish as a model system. Following is an overview of the major forward genetic approaches employing the zebrafish, along with examples that have provided fundamental new insights into vertebrate …show more content…
We will mutaganize a stock A population of N2 C. elegans, the P0 generation, will be acquired and mutagenized by Ethyl Methanesulfate (EMS). Individual worms from this generation will produce spatially separated F1 offspring. A select few F1 individuals will reproduce further and produce an F2 progeny.
● The F2 generation of this test group will then be screened for the roller mutation. The progeny of these rollers, the F3 generation, will yield the most integral data of our screen.
. These foundational worms will serve as our parent generation, P0. Once these worms have reproduced and eggs are visible on the plates, the P0’s will be moved to another 5 plates. This will ensure that we have ample offspring of the F1 generation in case issues such as contamination or starvation occur. Also, to safeguard against starvation, we will transport only 5-7 F1 worms to new plates to lay eggs, which will hatch to become the F2 generation. In addition to providing the worms with an ample food source, moving each successive generation of worms to fresh plates will allow us to more efficiently keep track of the offspring and the mutations observed in each generation. This is why it is imperative to maintain a strict schedule during the course of the experiment. Checking on the worms daily, we can see exactly when eggs are laid and when parent worms need to be removed in order to prevent mixing of
One containing wild type (WT) and the other having long (lon) C. elegans in order to identify males on each plate. Next, two new agar plates with E. coli as a food source for the nematodes were obtained and labeled with the phenotypes to be used for each cross. For plate 1, a single long hermaphrodite was found and with a heat sterilized worm pick, transferred to the new plate. From the WT parent plate, nine males were moved into plate 1 within close proximity of the hermaphrodite to help ensure fertilization. This cross was made because would show which is the dominant trait based. On plate 2, five long males were taken from the lon parent plate and placed along with a single wild type hermaphrodite from the WT parent plate. This cross was made to see if the trait was sex linked or not. This was followed by each plate being resealed with parafilm and given to the instructor to be put in storage until the following
Genetically transferring genes from one organism to another may sound like something in a sci-fi movie, but now it is happening more than ever. Snipping desired traits from various
The fruit flies were provided on March 27,2018. The fruit flies were incubated at 25°C for a couple of days. On April 3,2018 the F1 generation was examined which allowed to figure out the P generation by setting up crosses and collecting data. On April 10,2018 the identification, separation and counting of phenotypes in F2 generation was completed. On April 10, 2018 the fruit flies had to be counted correctly according to their phenotypes. Counting the sex of the fruit flies and analyzing for any specific mutation. On April 17,2018, F2 generation was counted and by that information that was obtained, a Chi-square test was made in order to prove or disapprove the hypothesis. Lastly, fruit flies were then removed from the cultured vials with larva and eggs and were placed in a sleeping chamber in order for the F1 generation to be counted and observed under the microscope. The reason why they had to be counted was to identify the sex and the phenotype from each individual fly. Then, the data was analyzed after the flies were counted, and a chi-square test was conducted. Since six vials were obtained from the instructor, two of the group members had to count 2 vials of F1 as well as F2 the other two members only had to count the fruit flies of one
The mealworm is the larva of the mealworm beetle which is a specie of darkling beetle (Animals.mom.me, 2016). The darkling beetle has a metamorphic life cycle that begins with an egg hatching into a small mealworm over a four week period (Sandhyarani, 2016). The second life stage of a darkling beetle is the larva stage where the egg will hatch into a small brown mealworm that eats and grows for about eight to ten weeks till its next stage as a darkling beetle pupa (Sandhyarani, 2016). The larva stage of a darkling beetle is the stage in which the mealworms will be investigated. The mealworm will then enter the pupa stage during one of its shell sheds causing the mealworm to
The experiment can further be improved by having an exact number of worms and E. coli on each agar plate to ensure every variable is the same except for the controls. Also, if the C. elegans were starved for a longer period they might have been more attracted to the diacetyl; further distinguishing the affected and unaffected worms (Urushihata et al. 2016).
Zebrafish (Danio rerio) are a popular freshwater fish that have been modified by researchers to produce several transgenic strains. The strain of transgenic zebrafish that express fluorescent colors have been given the name Glofish, for they contain different fluorescent proteins such as Green Fluorescent Protein (GFP), Red Fluorescent Protein (RFP), Yellow Fluorescent Protein (YFP), Blue Fluorescent Protein (BFP), or Purple Fluorescent Protein (PFP) (Vick, B. M., A. Pollak, C. Welsh, J. O. Liang, 2012). There are several benefits of using Glofish as a model to portray and understand inheritance patterns of transgenic genes. Glofish are small size in size, produce a high number of progeny, have similar genetic control to humans,
For the majority of animals on earth, responding to chemical stimuli could mean the difference between life and death. Taste is an example of sensing chemical stimuli-however, this response is not entirely dependent on genes, as tasting something will not necessarily trigger a gene to activate. Within a simpler organism however, such as Caenorhabditis Elegans, there is a complex behavior that is mediated by three sensory neurons and transduce chemical stimuli to move muscles that move an animal forward(attraction) or backwards(repulsion). By placing these organisms on several plates which will inhibit genetic expression, and then testing chemotaxis at a later point, it is possible to determine which genes are responsible for chemotaxis. The global sample averages of the chemotaxis indexes of CEH-36 and CEH-27 are somewhat inconclusive, with a mean of 0.5815 for CEH-36 and 0.6981 for CEH-27. Overall, the results for CEH-27 are likely to show that there is a lack of genetic interference with chemotaxis where CEH-36 is less definitive.
Zebrafish are an ideal organism to work with, they are easily stored and the cost value isn’t very high. The Zebrafsh that were preserved and used for trials in this research took place in the setting of the University of Sydney Animal Ethics Committee. “The fish were kept at a 14 h light/8 h dark cycle at 28.5°C “ (Rinwitz) There were two microliter of purified BAC DNA, which were mixed with Transpose mRNA. The transposes mRNA was shot into the zebrafish fertilized eggs. The following day, the zebrafish eggs were examined under the “fluorescent stereomicroscope” (Rinwitz). A fairly large mount off embryos survived to become adults. “Two to three positive founder fish were identified for each modified BAC and the F1 offspring was
Chronic pain, easy bruising, poor wound healing, premature rupture of membranes, and organ rupture were all listed above. Other possible complications include, but are not limited to, early onset arthritis and rupture of the eyeball (Ehlers-Danlos syndrome). While someone with EDS may experience life in a more challenging way, and potentially a more painful way, most types of EDS have a normal life expectancy. Vascular Ehlers Danlos Syndrome is the only type that has a shortened life expectancy due to sudden death related to spontaneous organ rupture (Ehlers-Danlos Syndrome). While Ehlers Danlos is still a highly-misunderstood disease, due to lack of funding and an overall anonymity of this disease. EDS is often considered an “orphan disease” due to its lack of funding and lack of research. While EDS will remain an “orphan” disease, there have been medical advances. There has been genetic test to find what the most common, hypomobility type, is cause by. Right not in Ghent, Belgian at the Ghent University Hospital there is a center called Centrum Medische Genetica Gent is researching this. They have conducted “first reported genome-wide linkage analysis in an EDS-HT family” (Syx, Symoens, Steyaert, De Paepe. Coucke, Malfiat, 2015) and finding the LZTS1 gene being connected to it. This type of research helps to provide hope for those suffering from this illness as new medical advances continue to be made because once a gene is discovered link to it medical care can be
Research of echinoderm regeneration on the molecular level has been taking a hit because of the lack of critical mass, but this has all changed lately. It is now possible for scientists to experiment echinoderm genome regeneration on the molecular level. The study done by this host of scientists from the University of London has one main goal, and that goal is to identify the genes that are involved in the regeneration process of Echinodermata. To be more specific, they are looking at the processes of neutral regeneration in specific echinoderm species. Echinoderms have been used as a model organism for development over a long period of time, but just recently they have been on the horizon of molecular advances.
Type II syndactyly or synpolydactyly(SPD) is a semi dominant inherited limb malformation that involves a fusion of digits. It is caused by mutations in HOXD13 on chromosome 2 due to polyalanine repeat expansions. Polyalanine repeats in SPD are mitotically and meiotically stable, causing polymorphisms to be rare, unlike other nucleotide repeat expansions such as Friedreich’s ataxia. HOXD13 is a member of the HOX family, a family of transcription factors that are proteins which contain homeodomain that are important for controlling cell fate along the limb axes and body. HOXD13 is a part of the HOXD gene cluster and crucial for limb development, particularly during the early and late stages of limb development. The stage occurs during the creation of the limb buds at week 4, during this stage the limbs have AP polarity through the expression of sonic hedgehog(shh) signaling from the zone of polarizing
Caenorhabditis elegans (C. elegans) are transparent roundworm, that range about 1mm in length and lack a respiratory and circulatory system. C. elegans is a model organism in genetics, its short life cycle, ease of cultivation, small size, and high cell number and development makes it an easy animal to use in experiments (International Journal of Molecular Science). In this experiment, two strains of worms will be used. Virgin hermaphrodite and him-8 males, which will be the wildtype. Him-8 strain is being use in this experiment because this gene has an increased rate of X-chromosomes, giving a forty percent change of producing male and sixty percent chance of hermaphrodite. This will help produce wildtype and mutant genes in the genetic offspring of the two. Hermaphrodites can be distinguishing from the male counterpart because they have wispy threadlike structure for their tail, whereas male have a blunt-end tail. (C. elegans as a genetic organism) The tails are a great way in telling the difference between these two mutant strain. When crossing these two strain together, the hypothesis that was made would be that we would find more male mutation of an offspring because the male strain is a more recessive gene trait. Dumpy A with a phenotype of dyp-10, this affects body shape and movement, causing the worm to be half of the normal length but the same width, which causes a stumpy effect.
In order to understand what a model organism is, we have to take a look into one of the most widely used organisms in genetics; Caenorhabditis elegans a nematode known for its male and hermaphrodite parts. Also known to be the most useful model organism because it is transparent, small in size and easy to cultivate. For this experiment we will be analyzing genes that regulate biological processes. The main focus will be what geneticists’ call the forward genetic screen, this technique is used to screen mutant animals for a certain phenotype. It was started by a man named Sydney Brenner who began with a certain phenotype and discovered mutants of that phenotype, essentially conducting a forward genetic screen. This technique is used to study genetic pathways that regulate biological functions, allowing scientists to focus on the pathway of the function of neurons, these neurons are what makes signals of muscle movement. This research can lead to better understanding of human genes and how to analyze pathways that are responsible for tissue and organ function. Sydney Brenner was able to identify over 600 mutants, for this lab we have focused on ~9 phenotypes, our given phenotype will be the Protruding vulva (PvL). Figure 1 is the C.elegans life cycle, the understanding of the nematodes lifecycle will allow for better results, knowing the time it takes for these worms to reproduce can provide us with the mutants we are looking for.