Protein extractions from unidentified fish samples were separated according to the molecular weights by SDS polyacrylamide gel electrophoresis. Since some of these proteins are shared between fishes, phylogenetic evaluation was reached. Western blot analysis was used to identify four unknown species of aquatic animals via comparison of actin/myosin bands. According to the results of this assay, the best estimate is that the unidentified aquatic animals are specimens of salmon, tilapia, cod, and shrimp, respectively.
Introduction
Western blot has been a revolutionary technique for identifying the expression of proteins within relative molecular biological samples that shared the same ancestor. Moreover, the sensitivity and specificity of the western blot (Immunoblotting) enables it a common technique for determining specific protein levels in clinical samples. Since the antibody specific to the antigen immunospecificity), it enables the target protein to be identified. Western blotting can produce quantitative data about that protein, which in this case the difference between bands in each of the protein samples. The western blot is an analytical technique used to detect specific proteins in the given sample of tissue homogenate or extract. The proteins are then transferred to a membrane (in this case, nitrocellulose), where they are stained with antibodies specific to the target protein [1] [2].
This experiment designed to analyze the phylogenetic of four unknown different fish muscles samples in order to predict the type of the aquatic creature. In this study, actin and myosin proteins were analyzed on account of their fairly high and constant expression in the animal muscles. Since the actin and myosin proteins are the two major...
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...and arthropod (shrimp/krill). Well number 7 has low protein actin and myosin and can be identified as shrimp, and wells 4, 5, 6 are identified as salmon, tilapia, and cod, respectively [3].
In figure 2, it is clear that the protein was transferred successfully from the gel to the membrane. The blot analysis was performed to detect if the protein was expressed.
The amount of protein present in each sample dilution in the blot corresponds with an absorbance value for that unknown protein extract. The antibody will bind to the protein (actin or myosin) and the Horse Raddish Peroxidase will catalyze the cleaving of the substrate to produce color upon antibody-protein binding. According to figure 3, 4, 5, 6 lanes are similar which confirm that they shared the same ancestor. However, lane 7 is slightly different which approves again it evolves from different ancestor.
Digestion of the haemolytic and non-haemolytic cells allowed for easier identification of fragments during electrophoresis analysis. Lane 12 in figure 3 show the size markers of SPP1 digested with EcoR1 while lanes 6 and 7 show samples of pK184hlyA and pBluescript digested with EcoR1 and Pst1. Lane 4 was loaded with plasmid DNA from haemolytic cells digested with EcoR1 and Pst1 while lane 5 was loaded with EcoR1 and Pst1 digested DNA from non-haemolytic cells. There was a lack of technical success in both lanes due to no bands appearing in lane 4 and only a single band appearing in lane 5. Theoretically, two bands should appear in both lanes after successful to allow for fragment identification. A possible explanation for the single, large fragment in lane 5 is that successful digestion did not take place and the plasmid was only cut at one restriction site leaving a large linear fragment of plasmid DNA. The absence of bands in lane 4 could be because there was not enough plasmid loaded into the lane. Another possibility could be that low plasmid yield as obtained when eluting the experimental samples in order to purify it. Lanes 8 and 9 belonged to another group and show technical success as two bands were present in both the haemolytic (lane 8) and non-haemolytic (lane 9) lanes. If the
completed, the tubes are stored at 4°C until analysis of the tubes. To alylize the PCR results with
The purpose of this experiment was to discover the specificity of the enzyme lactase to a spec...
Seahorses are a prime example of species whose atypical biology and unusual global distribution leads to a series of evolutionary questions. Seahorses (genus Hippocampus) are a marine species that have extensively been studied because of their abnormal behaviors in the marine environment compared to other marine creatures. Many of the seahorse species have large ranges, both longitudinally (over a great horizontal distance across the ocean), and latitudinal (great vertical distance within the ocean), regardless of the fact that they are characterized as weak swimmers and lack any large structural fins for efficient swimming (Lourie et al., 1999a). Although they do have these large range environments, seahorses
Acknowledgements: Slides of dead fish courtesy of OKDEQ. We would like to thank our students Trevor Nance Jr, and Matt Ward for their help in the laboratory sample preparations. We would also like to thank OKDEQ (Chris Armstrong) and EPA Region 6 (Rick McMillin) for their patience.
Scorpaenidae are mostly marine fish,very few found in fresh and brackish water. The family have around around 45 different genera and 380 different species. They are mostly found in the India Ocean and the Pacific Ocean. Most of them lives in coral, tropical reefs and shallow waters, warmer areas. Not all of the family fish lives in the shallow water, small portion of the Scorpaenidae live deep as around 7,000 feet. Just like most of the coral reef fish are, Scorpaenidae have camouflaged body. Their characteristic reflects their name, they have sharp spine that can sting other organisms. Their spine are covered with venomous mucus just like how some other kind of fish are covered with the venomous mucus. Their body are covered with scales. Generally speaking, the Scorpaenidae family fish have different number of spines over the course of the body. Most known type pf Scorpaenidae are lionfish and scorpionfish. Lionfish have preorbital bone with 3 spines, spines and third below the diagonal; , big mouth, end position, oblique fissure. Mandible, vomer and jaw bone with villiform teeth group. Gill openings are wide, tetrabranchiate membranes without the isthmus, pre-opercular margin with 5 spines. Body are ctenoid scales, round head. On the side, there are about eight to nine dorsal fin, fin spines and rays of an anal fin; five to six pectoral fins, they are large, round, and are branched from the upper fin rays. There are about one to five pelvic, sub thoracic; caudal fin rounded truncate. Reef stonefish are extremely camouflaged, they could look exactly like a rock when hiding for predation. The spines of Scorpaenidae are hard and rough, some of their fin spines have poison, can cause serious pain, swells and fatal wounds. Some of ...
Rekacewicz, P. (2008). Consumption of proteins from fish in % of total consumption of animal proteins. [Diagram]. Retrieved from
The origin of modern day whales, a mystery that has puzzled paleontologists for years, may have just been solved with the discovery of an ankle bone. This discovery might sound simple and unimportant, but the bones of these ancient animals hold many unanswered questions and provide solid proof of origin and behavior. The relationship between whales and other animals has proven to be difficult because whales are warm-blooded, like humans, yet they live in the sea. The fact that they are warm-blooded suggests that they are related to some type of land animal. However, the questions of exactly which animal, and how whales evolved from land to water, have remained unanswered until now.
Fish were amongst the first known chordates about 500 million years ago. Therefore, they have a very elaborate and complicated evolutionary history. The first type of fish to appear during the Ordovician era were called Ostracoderms. They had their head and flanks covered with a bony armor while they had a cartilaginous back. The shield was used to keep off predators. The extinction of the Ostracoderms during the Devonian period saw the emergence of Placoderms. These types of fish were also armored, but their functional jaws distinguished them from their predecessors. They also have gills derived from their jaws that took over the specialized role of gaseous exchange. Evolution of
There are various computer programs that can aid in sequencing these segments of DNA and generate phylogenetic trees. The programs are specifically looking for intron indels, retroposon, gene duplications and linked genes (Rokas and Holland 2000). Researchers can use several supermatrix formations that have already been created to pair up species (Gatesy et al. 2012). In our example, DNA was extracted, purified and then sequences. The computer program was looking for the presence or absence of transposon sites when compared to one of the supermatrix formations(Gatesy et al. 2012). Additionally, as the computer program runs the sequences gathered from the whale and hippopotamus the sequences will align allowing for additional differences and similarities in the genetic code to be found. After the genetic code has been sequenced, the computer programs can run programs to make phyogenetic
... produces can be measured. (Chesney and Folkman, 1999) A positive result is identified by a polymerase chain reaction and the presence of the specific antibody.
= Before conducting the experiment I would conduct a simple test for the protein by placing a sample of the albumen into a test tube and add biurett reagent. This contains copper (II) sulphate and sodium hydroxide.
In two-dimensional electrophoresis gel of isoelectric focusing is placed horizontally on an SDS gel , the electric field is applied and the protein macromolecules are separated acrodo with the isoelectric point and in terms of its molecular mass , providing a electrophoretogram that enables viewing and identification of many proteins at once .
Ichthyologists say that there are more than 32,000 species of fishes discovered on Earth so far which is more than all the amphibians, reptiles, birds, and mammals combined. Still, new species are being discovered almost every day. Large areas beneath the ocean surface still remain unexplored. It is predicted that there might be more species of fish which are hidden from the eyes of humans till now. All fish differ in sizes. A fish can be as big as a 51 ft long Whale/Shark or as small as an 8mm Stout infant fish. Common type of fish include; gold-fish, Drum, Spadefish, Trout, Skate, Thalla, Tuna, Hogfish, Wahoo, Grunt, Monkfish, Roughy etc. Some organisms which are considered to be a fish actually do not fall into the category of fish. Examples include; shellfish, cuttlefish, starfish, crayfish and jellyfish.