Sunday, January 26, 2020

Hydrolysis of Phosphomonoesters Study

Hydrolysis of Phosphomonoesters Study Introduction Site-directed mutagenesis was the primary basis of this portion of the laboratory directed toward determining the active site of the enzyme catalysis for hydrolysis of phosphomonoesters. The criteria was to turn the original sequence with Arginine 166 into Glutamine 166 through a point mutation. The primer location is located in the 401511 sequence of the phoA gene of alkaline phosphatase in E. Coli. The primer sequence is: 5- G CTG GTG GCA CAT GTG ACC TCG CGC AAA TGC TAC GGT CCG AGC -3’. The mutated primer sequence is: R166Q1 5’-G CTG GTG GCA CAT GTG ACC TCG CAG AAA TGC TAC GGT CCG AGC-3’. The reverse complementary mutated primer sequence is: R166Q2 5’-GCT CGG ACT CGG ACT GTC GCA TTT CTG CGA GGT CAC ATG TGC CAC CAG C-3’. A calculation for the melting temperature can be determined by the following equation: TM= 81.5 + 0.41 (%GC) – 675/N- % mismatch The TM value for Gln-166 was determined to be 85.95ËÅ ¡C. It is necessary for the TM value to be above 75ËÅ ¡C for successful mutation. Running on the null hypothesis, it would be clear that nothing would change for the kinetic parameters. If something were to change, the kinetic parameters for the Michaelis constant (Km), which determine the velocity of an enzyme as well as calculated to be  ½ the Vmax, would be hypothesized much higher after the mutation to glutamine since the mutation allows for 61% more accuracy from arginine’s CGG at 25% to glutamine’s CAG at 86%. The increase in the binding affinity creates the hypothesis that since it is bound closer together creating a stronger binding affinity, the maximal speed (Vmax) would be reduced since it does not require as much conversion between substrate and product. Mechanism of Alkaline Phosphatase Alkaline phosphatase (APase) is an enzyme located in the periplasmic space of E. Coli. The structure of APase, usually in the form of a dimer, shows two zinc metallonzymes and a magnesium ion in the active center. There is a bridging ligand with the protein Asp51 in the active center for the two zinc ions and the one magnesium ion. There are four phosphate oxygens; two phosphate oxygens form a phosphate bridge between the two zinc ions and the other two form hydrogen bonds with the guanidinium group of arg-166 (Coleman, 1992). The dimer is two identical subunits that each contain 429 amino acids. (Coleman, 1992). The most important amino acids located at the active site of APase are the four Cysteine residues represented as combined intrachain disulfides. The phosphorylated residue was Ser 102, which is phosphorylated during phosphate hydrolysis, to begin the nucleophilic attack on phosphorus. Each monomer contains a leucine rich environment of 22 residues. (Coleman, 1992). The first zinc ion is necessary in activating the leaving group of the ester oxygen in order for Ser 102 to phosphorylate (Coleman, 1992). The second zinc ion is necessary for the ester oxygen of the serylphosphate to begin activation of the phosphoseryl intermediate during hydrolysis of Ser 102. When the phosphoseryl intermediate is formed during the first zinc formation, a hydroxide is formed. (Coleman, 1992). Figure 1: this figure represents the monomer of alkaline phosphatase showing the zinc triad with magnesium in the active center. (Coleman, 1992). The proposed mutation as discussed, is mutating the arg-166 to glu-166. It is speculated that there will be no effect on the enzyme during the mutation. Arg-166 is located in the guanidinium group and is an electrophilic species while APase is being phosphorylated. Arg-166 is speculated to play a role in stabilizing the developing negative charge on the oxygen of the leaving group, pentacoordinate transition state, or help bind the phosphate group. As discovered byButler-Ransohoff et al, it is discovered that Arg-166 has no effect on the hydrolysis of phosphate monoesters and therefore will not be effected after mutation. Competent Cells Competent cells are used to help the DNA get into the cell. DNA is negatively charged and the calcium ions from the competent cells of DH5-ÃŽ ± cells are positively charged. The purpose of calcium chloride’s positive ions is to create an equilibrium between negative charge on the cell membranes phospholipid heads and the phosphate group on the plasmid DNA. This is where the genetic modification occurs. Therefore, they help by binding to the DNA and move it into the cell after the cell goes through a heat shock cycle. Competent cells have a very high rate for transformation efficiency. The proposed understanding of how this works, is that the calcium ions are positive and therefore weaken the electrostatic repulsion, which in turn weakens the cell walls. When the cell goes through heat shock, it causes the pressure to increase opening the cell and allowing the competent cells to bring the DNA into the cell by allowing pores to be created with the weakened cell wall. This forces the plasmid to become supercoiled so that it can pass through the pores created by the charge difference and heat shock. The competence comes from the cold bath of DH5-ÃŽ ± cells in calcium chloride which shocks the cells causing pores and then heat the cell to 42ËÅ ¡C for roughly 2 minutes, more than this could cause the cell membrane to denature and the pEK-154 mutated plasmid would become denatured. DH5-ÃŽ ± is a strain from E. Coli commonly used in laboratory practices due to it having the phoA- gene deficiency. This is necessary and desirable because it is easily transformed for creating competent cells. DH5-ÃŽ ± cells are used because they are a common strain in E. Coli and are commonly used for cloning. In addition, they promote stability and help improve the quality of the plasmid when using the Miniprep kit. (Dagert, 1979). A growth curve was created after incubation of the cells for competent cells. This was done to show the growth period during the lag phase is depicted by the highest point on the growth curve. This is taken to determine how much growth you have and is determined through a logarithm vs. time on a graph. The method used is optical density (OD) at 600nm to measure the transmittance on a spectrophotometer. If the value for the OD600 is high, the protection factor by a filter is lower and vise versa. This method is used to determine how much light is absorbed through the bacterial cells. Site-Directed Mutagenesis The sequence chosen for the point mutation from Arginine 166 to Glutamine 166 was sequenced in Ann Arbor. Site-Directed Mutagenesis synthesizes two complimentary oligonucleotides double stranded DNA template with glutamine 166 mutation, which is tagged with unmodified nucleotide sequences to a single stranded DNA template to allow mutation to occur and then reforms the new double strand DNA template during thermal cycling. When this occurs, the newly single stranded DNA template forms complementary strands with the enzymes and nucleotides to produce a higher result of strands. The purpose of site-directed mutagenesis is to make specific changes to the DNA sequence of a gene. Quik Change Lightening Mutagenesis kit provides a faster and more reliable insertion of the mutation with a simple three step method. The multiple cycles are subjected to a polymerase chain reaction (PCR) method. This allows multiple sets of mutated DNA to be produced within a short period of time. PCR works by heating and cooling the samples, since the double DNA is subjected to separation when heated it causes the srtands to separate allowing nicks in the template to occur. pUC-18 is a control plasmid used for determining if a mutation was successful and efficient. Figure 2: pUC-18 plasmid DNA genetically mutated to contain the LacZ gene as well as the ampicillin resistance gene. The polylinker cuts the section of the DNA by the Dpn1 restriction endonuclease creating a linear DNA strand allowing for the binding of the mutation of Arg-166 to Glu-166 to occur. pUC-18 is a circular double stranded DNA molecule. The reason pUC-18 was chosen as the control positive plasmid is because it was genetically manipulated to contain an ampicillin resistance gene as well as a ÃŽ ²-galactosidase enzyme known as LacZ. The lacZ gene is essential for DNA mutation because it contains a region to insert a polylinker, which recognizes the Dpn I restriction endonucleases during digestion causing the plasmid to become linear and bind to the mutated plasmid DNA that has also been cut with the Dpn I restriction endonuclease. A pWhitescript is used as the positive control plasmid used for mutagenesis. The control plasmid is combined with pEK154 plasmid containing the phA gene of APase and the vector double stranded DNA. The two oligonucleotide primers are the opposite ends of the vector. pWhitescript has the stop codon TAA inserted that stops the ÃŽ ²-galactosidase enzyme from producing. The two oligonucleotide control primers create a point mutation that turns the T residue of the stop codon to the C residue of the glutamine codon (Gln, CAG). That allows enzyme ÃŽ ²-galactosidase to be produced after pWhitescript is subjected to mutagenesis. There are two oligonucleotide with the mutation primer as discussed earlier. New DNA polymerase called Q1 enzyme was used for double stranded DNA template when combined with dNTP mix it extended the two-oligonucleotide primers as well as allowing the new plasmid DNA to construct. PCR Cycling parameters were used during Quik Change Site-Directed Mutagenesis for the transformation of arginine to glutamine by the use of the Pfu Enzyme. As visible from table 1, the cycling parameters were set up for this specific reaction for a point mutation. After cycling, the non-mutated parental supercoiled double stranded (dsDNA) is digested. Table 1: Quik Change Site-Directed Mutagenesis Parameters Segment Cycles Temperature Time 1 1 95C 2min 2 18 60C 60C 68C 20 sec 10 sec 30 sec/kb of plasmid length (3min) 3 1 68C 5 min The first step of the three step simple method for Quik Change Lightening Mutagenesis kit is Thermal cycling at 95ËÅ ¡C for two minutes. During this time, the DNA template becomes denatured allowing the primers to be subjected to heat treatments, which denature the pEK-154 template DNA strand and synthesizes the primers to extend linearly and cause nicks with Pfu Fusion-based DNA polymerase, which are then sealed by components within the Pfu enzyme blend. This polymerase allows for exact replication of original template and does not disrupt the orientation. The original mutated pEK-154 was used as the supercoiled double stranded DNA template Figure 1: during thermal cycling, the double-stranded DNA Template is nicked and subjected to point mutation for arginine 166 to glutamine 166 and then sealed with components in the Pfu enzyme blend. In the second step of the kit the template digests the parental DNA strand with methylated and hemimethylated DNA using the enzyme Dpn I endonuclease and subjected to thermal cycling for the second segment. The un-mutated pEK-154 plasmid DNA is the parental DNA while the mutated pEK-154 is the plasmid DNA with the desired mutation. With the nicks, the mutated plasmid with the two oligonucleotide primers is mixed with Dpn I endonuclease for methylated DNA that targets the sequence 5’-Gm6ATC-3’. This sequence helps digest the pEK-154 parental DNA template and has not been introduced into the methylase enyme allowing methylation to not effect the DNA and therefore will not be digested with the parental DNA strand. Similarly, the un-mutated is also digested in Dpn I restriction endonuclease to be methylated. Figure 2: Dpn 1 enzyme endonuclease digests the parental DNA template with methylated and hemimethylated allowing for single stranded DNA for transformation. Step 3 is the last cycling step where transformation of the newly mutated glutamine 166 single-stranded DNA. The DNA is added to Dpn I restriction endonuclease, which forms it into linear DNA stranded DNA duplex forming a double stranded DNA for the newly synthesized mutated DNA. After the mutated vector DNA with the primers has been nicked it is transformed into CaCl2 competent DH5-ÃŽ ± cells. This transformation must occur to repair the nicks caused during cycles to separate the DNA strands. The pWs, which is used as the control DNA is also transformed with competent DH5-ÃŽ ± cells to help repair the nicks. Transformation Control screening used X-gal and IPTG to determine if ÃŽ ²-galactosidase activity. The newly mutated pEK-154 cells are screened for alkaline phosphatase activity. This helps screen for ÃŽ ²-galatosidase enzyme in E. Coli is a Lac Z gene that codes for this enzyme. This enzyme is useful because it breaks the lactose into galactose and glucose. A Lac operon contains an operator and a promoter, which binds RNA-polymerase that starts transcription for the Lac I gene for I protein which can only bind to operator or lactose, but not both. When I protein is bound to lactose, in this case Isopropyl-1-ÃŽ ²-D-galactopyranoside (IPTG) which mimics the structure of lactoase allowing RNA-polymerase to bind to the promoter to produce ÃŽ ²-galactosidase. 5-bromo-4-chloro-3-indolyl-ÃŽ ²-D-galactopyranoside (X-gal), which is a substrate that cleaves galactose in ÃŽ ²-galactosidase to produce a blue color. If no ÃŽ ²-galactosidase is present they turn white on the agar plates. This determines if the cells contain the mutated CAA codon for glutamine instead of the stop codon TAA of the unmutated protein. 5-bromo-4-chloro-3-indolylphosphate (BCIP) is used to determine if alkaline phosphatase is present. BCIP like X-gal is a substrate that cleaves the phosphate group off of alkaline phosphatase producing a blue color.

Saturday, January 18, 2020

Comparative Paper of Race/Ethnicity Essay

Racial background and ethnicities are represented in the short stories â€Å"Country Lovers†, â€Å"The Welcome Table†, and the poem â€Å"What It’s Like to Be a Black Girl†. All of these stories have a main character or protagonist black female. All three of these women deal with some degree of discrimination because of their color. The hardships that these women suffer during their life can be suffered by anyone but growing up in a discriminatory situation creates a more dramatic story. The main themes in â€Å"Country Lovers† are love and racial politics. Country Lovers was written during a time when Africa was suffering from racial segregation. This story has irony throughout the entire story. Thebedi and Paulus grow up together and they fall in love. They grew up in Africa during the apartheid when their country did not allow interracial relationships. Paulus Eysendyck was the son of the farm owner and Thebedi’s father worked on that farm. They both knew they could not be together publicly. During the apartheid in Africa it was illegal to have an interracial relationship. There are several dramatic effects in this story. The first is when the narrator talks about Paulus going away to school â€Å"This usefully coincides with the age of twelve or thirteen; so that by the time early adolescence is reached, the black children are making along with the bodily changes common to all, an easy transition to adult forms of address, beginning to call their old playmates missus and baasie little master† (Clungston, 2010). There’s loss of innocence and forbidden love as described here when Paulus watches Thebedi wade in the water â€Å"The schoolgirls he went swimming with at dams or pools on neighbouring farms wore bikinis but the sight of their dazzling bellies and thighs in the sunlight had never made him feel what he felt now when the girl came up the bank and sat beside him, the drops of water beading off her dark legs the only points of light in the earth–smelling deep shade† (Clungston, 2010). This love would by any other means be normal, but since it is during the apartheid it is against the law. Eventually, Thebedi becomes pregnant at eighteen with Paulus’s child. In order to protect herself Thebedi marries another man, Njabulo a laborer on the Eysendyck farm, like her father. When Paulus returns home on holiday he learns of the child, fearing that it is his, knowing the legal issues he could face, he goes to see the child. When Paulus sees the child â€Å"He struggled for a moment with a grimace of tears, anger, and self–pity. He said, â€Å"You haven’t been near the house with it? † (Clungston, 2010) Both Paulus and Thebedi know the consequences if the child is found out about. Two days later Paulus returns to Thebedi’s hut and drowns the child. The baby had been given a proper burial until â€Å"someone—one of the other labourers? their women? —had reported that the baby was almost white, that, strong and healthy, it had died suddenly after a visit by the farmer’s son†(Clungston, 2010). In the end, a trial resulted in a â€Å"not guilty† verdict because of insufficient proof. Each one of these events is dramatic. The main themes of â€Å"The Welcome Table† are impartial Christ-like love and racism. Walker’s story â€Å"The Welcome Table† never mentions a table except under the title it quotes an old spiritual. We are never given a name of the old woman in this story. This creates anonymity about the woman; this is tragic because she is unknown. Based on the description of the woman’s clothes the idea is given that â€Å"Perhaps she had known suffering â€Å"(Walker, 1973). In the story of the old black woman is described as, â€Å"the color of poor gray Georgia earth, beaten by king cotton and the extreme weather† (Walker, 1973). This old Black woman is on a mission. Even though there is no table in this story, the welcome table is a metaphor for impartial love. The old woman heads into a house of god expecting it to have impartial love. The church people discriminated against her because she is black. The good church folk are shocked. The reverend reminds her gently saying â€Å"Auntie, you know this is not your church† (Walker, 1973). The old woman thinks â€Å"as if one could choose the wrong one† (Walker, 1973). She brushes past them all and finds a seat near the back. Inside it is very cold, colder than usual. She ignores the request of an usher, referring to her as grandma, who asks her to leave. The ladies, who are celebrating the impartial love that they presumably have, finally insist and their husbands hurl her out. She is stunned, bewildered, and starts to sing a sad song. Then she notices something coming down â€Å"the long gray highway. † She grins toothlessly and giggles with joy. For it is none other than Jesus, and he is walking toward her. When he came close, he said, â€Å"Follow me† and the old woman â€Å"bounded down to his side with all the bob and speed of one so old†(Walker, 1973). The two of them walk on together. She tells him her troubles, and he listens kindly, smiling warmly. Jesus provides her with the welcome table. The people in the church never knew what happened to her. Some said they saw her jabbering to herself and walking off down the highway all alone. â€Å"They guessed maybe she had relatives across the river, some miles away, but none of them really knew. † The theme in this story is racism and hardship. Smiths poem gives the audience a view into a young girls transition from being a black girl into becoming a black woman during a time when both being a black girl and a black woman are unwelcomed. An Explication: From transition to disappointment. The poem â€Å"What it’s like to be a Black Girl (for those of you who aren’t)† by Patricia Smith, is exactly as it is described in the title. Smiths poem gives the audience an insider’s view into a young black girl’s transition into black woman-hood at a time where both being a black girl and a black woman was not as welcomed. Puberty is usually described by the biological changes a young boy or girl’s body goes through. Smith writes, â€Å"It’s being 9 years old and feeling like you’re not finished,† and â€Å"like your edges are wild, like there’s something, everything, wrong† (Smith, 1991). Though all teens have these thoughts in Smiths poem the black girl also have the added pressures of a racially unjust society. This â€Å"black girl† she refers to in her poem is feeling the awkwardness of her newly changing body and the hope of something different and maybe better to come. The poem tells the story of a young black girl exploring and experiencing what it is to become a black woman in a society that tells her to be white is better. â€Å"It’s dropping food coloring in your eyes to make them blue and suffering their burn in silence. It’s popping a bleached white mophead over the kinks of your hair and primping in front of the mirrors that deny your reflection. † (Smith, 1991) The food coloring in her eyes and the bleached hair symbolize her need to be accepted by society’s idea of proper. â€Å"It’s flame and fists and life according to Motown† (Smith, 1991). The life she knows is Motown music, racial slurs, and fighting. Between â€Å"jumping double Dutch until your legs pop† and â€Å"growing tall and wearing a lot of white† (Smith, 1991) the poem tells us how a young black girl balances her changing body, with her child-like mentality. The wearing of a lot of white is her wearing of the wedding gown often seen as a symbol of womanhood. On that day, she’s starts the next chapter in her life, as a married woman. When Smith talks about â€Å"having a man reach out for you and caving in around his fingers† (Smith, 1919) it gives the reader a better observation of the subservient mentality women dealt with during the 1960’s. Finally, this young black girl is now a woman. Throughout the poem, Smith has helped us to see the transition from a black girl to a black woman. With Smiths’ attention to detail, the reader is able to follow the girl’s changes, both biological and psychological. This poem tells the story of a young black girl’s journey and her experiences while becoming a grown black woman in an era of racial uncertainty. All three women are survivors of a life of racial unjust. These stories are common to everyday life changes and lessons. These hardships, that everyone normally sees, are much more dramatic in a society that discriminates against color. References: Clugston, R. W. , (2010). Journey into Literature. San Diego: Bridgepoint Education, Inc. Retrieved March 20, 2011 from https://content. ashford. edu/books/AUENG125. 10. 2/sections/h3. 2? search=Country%20Lovers Walker, A. , (1973). In Love & Trouble: Stories of Black Women. New York: Harcourt Brace Jovanovich, 81-87. Smith, P. , (1991). Life According to Motown. What it’s Like to Be a Black Girl (for those of you who aren’t). Tia Chucha Press.

Friday, January 10, 2020

Cloudstreet Essay

Cloudstreet: Religion and Spirituality | A Novel by Tim Winton| â€Å"From separate catastrophes, two rural families flee to the city and find themselves sharing a great, breathing, shuddering joint called Cloudstreet, where they begin their lives again from scratch. For twenty years they roister and rankle, laugh and curse until the roof over their heads becomes a home for their hearts.† (Winton, 1991) Tim Winton’s critically acclaimed novel, Cloudstreet is a masterful tale of love, meaning and heartbreaking tragedy that speaks strongly of a post war Australian society that was essentially rebuilding itself after years of political upheaval and financial struggle. Good Morning/Afternoon Ladies and Gentleman. I am a representative of the National English Curriculum board and today I am here to demonstrate to you how Cloudstreet is authentic and believable, and as Marieke Hardy suggests: â€Å"It is Australian. Reading it felt like coming home.† Throughout Cloudstreet, there are numerous concepts that portray the Australian cultural identity; and the theme of religion and spirituality is especially prominent and appealing. The concept of luck, Aboriginal spirituality, and the search for the meaning of life, are all Australian ideas that Winton expertly portrays. Spirituality can be defined as â€Å"a concern for that which is unseen or intangible; as opposed to physical or mundane.† (Greenberg, 2008) It encourages a sense of peace and purpose within an individual and promotes a feeling of belonging. Additionally, religion can be defined as â€Å"the belief in, and worship of a superhuman controlling power.† (Religion) Both concepts are widely integrated into the core of the novel and are depicted through the Australian notion of luck. Luck, which some would argue has long been etched into the Australian consciousness as a common working class superstition, is, whether th ey are conscious of it or not, a form of religion for both families. The Pickles family, most notably Sam, rely on the â€Å"shifty shadow of God† (p 12) to warn them about future events, while the Lamb’s simple game of â€Å"spinning the knife† (p 53) acts as their metaphorical life compass. â€Å"The Lucky Country† (Horne, 1964) is a phrase that originated from a book of the same name written in the 1960’s, and since then, has gained widespread popularity and thus, been attached to the Australian culture for a long time. Winton has  cleverly examined this historical background to incorporate an accurate facet of the Australian identity into the novel and its characters. Also related to the concept of luck, is the fact that after Fish drowns, Oriel, once a devoted and â€Å"god fearing† Christian, begins to question her faith and the reliability of believing in God. When Fish is resuscitated, but only â€Å"some of him comes back†, (p 32) both she and Lester are emotionally forced to abandon God and Christianity and instead, turn to luck, hard work and the idea that â€Å"life and death, was all there was,† (p 65) in order to endure their circumstances. This draws on the common â€Å"Aussie battler† tradition, of which a working class person overcame challenging situations through perseverance, faith and steadfast determination. In terms of the Australian cultural identity, Winton has again taken an important and recognized historical Australian idea and shaped it to evoke feelings of familiarity and intimacy between the readers and the characters of Cloudstreet. The frequent appearance of the â€Å"Blackfella† is yet another example of how the Australian cultural identity is portrayed through examination of Aboriginal Spirituality. However, in many scenes throughout the novel, the blackfella signifies both Christian and Aboriginal spirituality through allusion and comparison. For example, he is likened to Jesus by walking on water and again when he produces a never ending supply of wine and bread in Quick’s car. This comparison is particularly effective as it symbolises the â€Å"coming together† of Christianity and Aboriginality, which was a particularly delicate Australian issue during the time period of the novel, due to Aboriginal marginalisation and the rise of Christian ideals. Essentially, the Blackfella acts as a reminder of the original religion inherent to Australia and its development, during a time when social and political change was overtaking that of its native beliefs. The â€Å"Blackfella† also ac ts as the conscience of the characters when they have lost their way or their family unit is threatened. This can ultimately be seen when he leads Quick back to Cloudstreet after he runs away to the country, knowing that Quick feels secretly lost without his family, and needs them to feel fully alive. He also persuades Sam not the sell the house and states that â€Å"you shouldn’t break a place. Places are strong and important,† (p 406) referring to not only the house and its tragic Aboriginal history, but also to the fragile  families who live inside it. In doing so, he ensures that the families stay whole and together, which is an important and dominating religious value for Aboriginality and Christianity, both during the time period of the novel and in our modern Australian society. Consequently, the â€Å"Blackfella’s† role in Cloudstreet is a significant contribution to the novel’s relevancy to the Australian cultural identity. The Australian cultural identity is also illustrated in Cloudstreet through the spiritual symbolism and personification of the river, and its connection to the character’s search for the meaning of life. This is particularly significant for Quick Lamb, who, is spiritually linked to the river in a number of ways. The river acts as a place of peace, purpose and belonging for Quick. Connecting with his mother when they go prawning, glowing after fishing in the country, and most importantly, falling in love with Rose Pickles, are the most significant spiritually defining events that Quick experiences while on the river. Through realising just how symbolically important the river is to him, Quick finally understands the true meaning of his life, and gains a feeling of belonging that allows him to finally shed his self-degrading title of â€Å"the lost lamb.† (p 310) Australia is a country that values the water. Geographically, we are surrounded by it, with most of our population residing close to the shores. As a result of this, the water is seen as a common gathering place, from which one cannot easily escape nor regard as irrelevant to the Australian way of life. Winton has taken this idea and incorporated it into Cloudstreet, to emphasise and promote a relevant part of Australian culture. Finally, the river’s spiritual and religious connection to Fish Lamb is perhaps the most important concept of the novel. After Fish drowns and has his soul ripped into two separate pieces (spiritual fish and physical fish), the river that he so desperately longs for, essentially becomes his gateway to the spiritual world; to the place where he belongs. It is not until the end of the novel when Fish is finally free to reunite with the water that he is truly whole again. â€Å"I burst into the moon, sun and stars of who I really am. Being Fish Lamb. Perfectly. Always. Everyplace. Me.†(p 424) For many, water in Australia is culturally considered to be the blood of the country; a place of c leansing and rejuvenation. Likewise, for Fish, the river embodies the epitome of the spirit of Australia in the form of life giving water. Although his life was initially taken by the water, it is eventually returned to him when his physical self re-joins his spiritual self. In conclusion, Winton flawlessly encapsulates the cultural identity and spirit of Australia in Cloudstreet through symbolic representations of luck, Aboriginal spirituality and the search for the meaning of life. The characters’ connection with religion and spirituality resonates strongly with the reader and successfully evokes feelings of belonging and familiarity that confirms Cloudstreet is indeed a classic Australian novel. Bibliography Associates, R. Q. (2008, September 16). Ideology in Cloudstreet . Retrieved 2013, from www.englishcurriculum.com.au Cloudstreet Notes. (n.d.). Retrieved 2013, from Sydney Home Tutoring: http://www.sydneyhometutoring.com.au/wp-content/uploads/2013/02/Details-Cloudstreet-notes.pdf. Dot Point Notes Cloudstreet. (n.d.). Retrieved 2013, from Sydney Home Tutoring : http://www.sydneyhometutoring.com.au/wp-content/uploads/2013/02/Dot-Point-Notes-Cloudstreet.pdf. Greenberg, N. (2008, October 8). Retrieved 2013, from Can Spirituality Be Defined: http://notes.utk.edu/bio/unistudy.nsf/935c0d855156f9e08525738a006f2417/bdc83cd10e58d14a852573b00072525d Horne, D. (1964). The Lucky Country. Penguin Books Australia. Religion. (n.d.). Retrieved 2013, from Google Definitions: https://www.google.com.au/search?q=religion+definition Winton, T. (1991). Cloudstreet. McPhee Gribble.

Thursday, January 2, 2020

The Life and Legacy of Dr. Kenneth B. Clark The History...

To address the concerns of social justice, while Clark was in office, he helped develop the Board of Social and Ethical Responsibilities of Psychology in 1972 (Pickren, 2002). Though Dr. Kenneth Clark has contributed much more to the world of psychology, he most importantly opened up and investigated racism and the psychological effects it has on children and has helped change the face of education in the process. Dr. Kenneth B. Clark’s legacy has lived on and will continue to inspire because, even today, in the 21st century, there are many ideas and problems that Clark addresses in the realm of prejudice and racism that are still relevant in social identity, education and the work place in America. Clark was a social psychologist†¦show more content†¦Clark’s theories of racial progression are still at work today and will continue to have a long lasting effect on America. Clark’s work on identity and stereotyping has continued and has extended to not onl y minority groups, but to white-Americans and women as well. Research on prejudices and stereotyping has continued in the area of education. According to Steele (2004), the framework of one’s life can be controlled by their social identity and that individuals must contend to the prejudices and stereotypes that they socially must identify with. Further research suggests that those who are exposed to negative stereotypes are influenced either by believing or mimicking the stereotype or, both (Steele, 2004). Steele (2004) found that stereotypes have contributed to the underperformance of minority groups in the school setting. As suggested by Clark, Steele (2004) also believes that conforming to suggested stereotypes weakens the social identity and brings severe deficits to the ability of student to excel in the classroom. Clark’s ideas on stereotyping were elaborated in the notion that stereotype threat does not only affect minority groups, but can exhibit deficits in a ny group of individuals. For instance, Steele (2004) offers the example of white vs. black athletes and how in many sports (e.g. basketball or football) white athletes have shown to feel less competent in comparison to black athletes because ofShow MoreRelatedStephen P. Robbins Timothy A. 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