Preview

Mr.Raviteja

Better Essays
Open Document
Open Document
7097 Words
Grammar
Grammar
Plagiarism
Plagiarism
Writing
Writing
Score
Score
Mr.Raviteja
“CLONING OF HORSE RADISHPEROXIDASE GENE IN E.coli FOR EXTRACELLULAR EXPRESSION”

Project report submitted to
Jawaharlal Nehru Technological University Hyderabad in partial fulfillment for the award of degree of
Bachelor of Technology in Biotechnology

Submitted by

M.RAVI TEJA
(08311A2338)
&
V.MAYURI
(08311A2358)

Under the Guidance of

DR.CHAND PASHA

Department of Microbiology
Osmania University

[pic]

Department Of Biotechnology
SREENIDHI INSTITUTE OF SCIENCE AND TECHNOLOGY Yamnampet, Ghatkesar (M), R.R. Dist., Hyderabad - 501301.
Andhra Pradesh

Department of Biotechnology
Sreenidhi Institute of Science and Technology
Yamnampet, Ghatkesar, R.R.District-501301

[pic] CERTIFICATE

Certified that this is the bonafide project report of work carried out by Mr. M. RAVI TEJA (08311A2338) and Ms. V. MAYURI (08311A2358) at Department of Microbiology, Osmania University. The work incorporated in this report entitled “CLONING OF HORSE RADISH PEROXIDASE GENE IN E.coli FOR EXTRACELLULAR EXPRESSION”.

Internal guide Head of Department

External Examiner

|Contents |Page No. |
| | |
|Declaration | |
| | |
|Certificate | |
| | |
|Abstract | |
|



References: M.RAVITEJA (08311A2338) V.MAYURI (08311A2358)

You May Also Find These Documents Helpful

  • Good Essays

    Plasmids are small circular autonomously replicating pieces of DNA that can be found inside of a prokaryotic bacterial cell. By barrowing a cell’s polymerase they replicate their own DNA. They are easy to extract from the bacterial cells due to their size. Plasmids are helpful for cloning foreign genes because of their ability to express antibiotic resistance as well their ability to be modified to express proteins of interest. A pGLO plasmid contains genes for the green florescent protein (GFP) as well as the gene for ampicillin resistance known as beta-lactamase. It also contains a gene regulation system (operon) that has the ability to control expression of the GFP gene in transformed cells known as araC. The source of GFP is naturally founds within a…

    • 463 Words
    • 2 Pages
    Good Essays
  • Good Essays

    Peroxidase is an enzyme found in potatoes that catalyzes the breakdown of hydrogen peroxide, H2O2, into O2 gas and water. We examined the different pH environments that can affect the enzyme activity during the breakdown of H2O2. In order to do this, we added different levels of pH, low, medium, and high, into different test tubes with the enzyme and H2O2, and we then inverted the tube. The amount of O2 gas produced was then measured and recorded. The result was that the higher pH produced more gas, followed by medium pH, then low pH. The enzymes were more active in the pH of about 10. It increased the enzyme activity allowing more O2 to be produced from the breakdown of the hydrogen peroxide.…

    • 621 Words
    • 3 Pages
    Good Essays
  • Better Essays

    Peroxidase Enzyme Lab

    • 805 Words
    • 4 Pages

    The hypothesis for the effect of various pHs on the activity of the peroxidase enzyme was not clear enough and did not predict any results. Based on the experiment, the best pH for the peroxidase enzyme is pH 5.…

    • 805 Words
    • 4 Pages
    Better Essays
  • Satisfactory Essays

    From the data collected in the experiment the hypothesis that was proposed was not supported. The hypothesis was that after the addition of a lower pH, or an increase in temperature the reaction would proceed faster. The results shown from the experiment are in accordance with the principles of enzyme activity. The varying environmental factors tested changes in pH, temperature, and concentrations of the enzyme affected the activity of horseradish peroxidase. The changes in pH decreased the enzyme activity the further it moved away from pH 7, the optimum pH. Changes in the pH first affect the form of the protein, hydrogen bonds between the amino acids of the molecule and progress until the form of the active center of the enzyme.…

    • 181 Words
    • 1 Page
    Satisfactory Essays
  • Better Essays

    In this experiment we tested the effects of the environmental parameter pH on peroxidase activity. We performed this experiment to observe and analyze the effect pH has on peroxidase activity and find the optimal pH level for peroxidase, as well as the ceasing point in the reaction. Parameters are characteristics that can alter an experiment. The parameter we used in this experiment was pH. The pH scale ranges from levels 0 to 14, 0 being the most acidic, and 14 being the most basic. An enzyme is a biological catalyst made up of proteins. Enzymes speed up reactions by bringing reactants close together and lowering the activation energy. Enzymes are extremely specific to their substrates. A substrate is material where an organism lives or the surface upon which an organism grows or is attached. Gauicol was the substance we used as a reducing…

    • 1598 Words
    • 7 Pages
    Better Essays
  • Good Essays

    Peroxidase Experiment

    • 1061 Words
    • 5 Pages

    Over a two week period of time in the laboratory, we experimented and tested the reaction rate of a peroxidase enzyme and the factors that affected it, both positively and negatively. The purpose of these experiments was to probe and manipulate the activity of the enzyme peroxidase by varying temperature, pH, the amount of enzyme compared to the substrate and the effect of hydroxylamine. Peroxidase activity is expressed when the potato extract is subjected to stresses such as low temperature (El-hilali et al., 2012). The most eye catching factors that we tested for their impact on enzyme activity involved change in pH, temperature, boiling extract, and the effects of probing the active site with hydroxylamine. In the first part of…

    • 1061 Words
    • 5 Pages
    Good Essays
  • Good Essays

    Mr.Roro Yazzzz

    • 445 Words
    • 2 Pages

    The Wicker Bar is a pompous place that normally features two French girls singing dirty songs, though the girls don't seem to be there when Holden arrives to meet Carl Luce. Carl, who is a bit older and certainly considers himself in another planet than Holden as far as wisdom goes, comes across as a pretty stiff and snobby guy. He drinks extra-dry martinis and dismisses most of Holden's attempts at conversation, which are admittedly a bit juvenile. Holden is particularly agitated about sex in this scene, and has his first bout of homophobia, calling gay men "flits." Carl Luce apparently knows some things about girls and sex. He says, for instance, he's currently dating a foreign sculptress in her late thirties. Holden keeps trying to get some details out of him. Carl Luce refuses to answer these "typical Caulfield questions...." (pg. 146) When Holden tries to be honest and explain how the problem with his sex life is that he can't get turned on by a girl unless he really likes her, Carl advises he go to a psychoanalyst. Luce's father is a psychoanalyst, and apparently Carl has made this suggestion before.…

    • 445 Words
    • 2 Pages
    Good Essays
  • Better Essays

    Peroxidase Lab Report

    • 1858 Words
    • 8 Pages

    The enzyme, peroxidase, extracted from a turnip was tested for its efficiency in binding to its substrate and its stability under several conditions. To do this, we tested effects on peroxidase activity, first, with different amounts of the enzyme, next at temperatures of 4oC, Room Temperature, 32oC, 48oC and boiling; then, at pH 3, pH 5, pH 7 and pH 9; and, finally, with the competitive inhibitor, hydroxylamine. We were able to measure enzymatic activity by the change in absorbance per second with a spectrophotometer. By testing different concentrations of peroxidase and its reaction rate in seconds, we were able to see that as the amount of enzyme increased the catalytic reaction also increased. The optimal amount of peroxidase concentration to be used in the subsequent experiments was determined to be 1.0 mL. Any amount above this would have caused the rate of absorbance to be too fast, making it too difficult to get accurate readings. Any amount below this would not have produced a reaction “at an appreciable rate.” (Dolphin, Vleck, Colbert and Westgate, p. 76) In addition, our results show that a rise temperature and pH only increase the rate of reaction to a certain point before the reaction rate begins to decline dramatically. In the case of boiling of the enzyme there was no rate of reaction found whatsoever. A similar result was found when hydroxylamine was added to the peroxidase and it caused an inhibition reaction. Overall, the results show that the peroxidase enzyme is sensitive with reference to the above factors in whether or not a reaction is catalyzed.…

    • 1858 Words
    • 8 Pages
    Better Essays
  • Good Essays

    In the story The Scarlet Ibis, Brother tried to teach Doodle to walk in order not to make him embarrassed.He finally succeeded. Later, he wanted more and asked doodle to learn to swim, to run and to do everything that normal kids could do. So it comes to the question: is it is the brother’s fault that caused the sad ending and do the motives matter in the end though you are doing something good?It looks conflicting in the story. Brother taught Doodle to walk with selfish motivation, but he succeeded and everyone was happy. Later, he tried to teach Doodle more with the motivation to make Doodle a normal kid .But he failed and Doodle died. The story did not give an exact answer .But it suggested that the motives do not matter so much to the result as we always regarded. There are good motivations driving us to do good things. However, the motivation from desire and interests are from human nature and are always strong. It is a respectful but…

    • 473 Words
    • 2 Pages
    Good Essays
  • Good Essays

    Chitobiase

    • 380 Words
    • 2 Pages

    In this experiment, a restriction map for restriction enzymes Eco R1, Pst1 and Hind III using Southern hybridization and restriction analysis of pRSG 192. pRSG 192 is a recombinant plasmid derived from the chb gene and pUC 19, a 2.7kb engineered plasmid which encodes for ampicillin resistance, a portion of the lac operon and a multiple cloning region . The chb gene exists as a 3.6 kb insert in the mutiple cloning region of pUC 19.…

    • 380 Words
    • 2 Pages
    Good Essays
  • Good Essays

    Mr Namdar

    • 748 Words
    • 3 Pages

    Belonging can be understood as having a place in the society. Belonging is influenced by social, historical, cultural and personal experiences, which all help to shape our identity. Our choices can lead us to feeling like we belong, or there may be barriers preventing us as individuals from making connections to people or groups. Belonging gives us security and ensures us that we belong to place, family, and culture. While not belonging can lead to insecurities which leads to questioning yourself if your good enough for the society to belong.…

    • 748 Words
    • 3 Pages
    Good Essays
  • Good Essays

    The Great Depression of the 1930’s was devasting. It deprived many people jobs, land and their livelihoods. It began in the United States and quickly spread to other parts of the world. Many people lost everything and were living on the edge with nothing but scrapes of food from dumpsters and occasional soup kitchens to keep them alive. People blamed themselves for their loss of jobs, so it became an epidemic.…

    • 587 Words
    • 3 Pages
    Good Essays
  • Good Essays

    Gene Cloning

    • 1802 Words
    • 8 Pages

    One aspect of the DNA cloning experiments that is carefully considered is the selection of cloning vectors. A variety of vectors have been created, each being suitable for a particular use. One common vector used in laboratories is a plasmid called pUC19. It is 2686 base pairs long and possesses an origin of replication which allows the production of over 100 copies in a competent E.coli cell. It possesses a multiple cloning site (MCS) which is artificially implanted by adding a polylinker sequence to it. The pUC19 plasmid is also altered by inserting a gene that codes for beta-lactamase which confers resistance to the antibiotic ampicillin (Read and Strachan 2011). The MCS occupies the 5’ end of the gene lacZ (Sherwood, Willey and Woolverton 2012). This gene codes for only the alpha-peptide of beta-galactosidase, an enzyme used to break down the disaccharide lactose into glucose and galactose (Read and Strachan 2011). The aim of this experiment is to incorporate a cDNA called CIH-1, from plasmid pBK-CMV, into pUC19.…

    • 1802 Words
    • 8 Pages
    Good Essays
  • Good Essays

    Mission Trip To Haiti

    • 1425 Words
    • 6 Pages

    As I read the book Strength in What Remains and watched the film God Grew Tired of Us, my heart broke for the people suffering from wars and poverty in other countries around the world. As Deo described the suffering in his village, I was brought back to my mission trip to Haiti, where my eyes were truly opened to the world of poverty for the first time. I went into the trip anticipating it would affect my life, but I had no idea how much I would truly be changed.…

    • 1425 Words
    • 6 Pages
    Good Essays
  • Better Essays

    Mr.India

    • 1186 Words
    • 5 Pages

    The is various interactions between the characters that illustrate the aspects of belonging in the community.…

    • 1186 Words
    • 5 Pages
    Better Essays