Starch is polysaccharide which play important role in the metabolism of living organisms. In addition, starch are raw materials in the food and materials industry. Increasing the starch content of crops based on improving the activity of some genes encoding enzymes plays an important role in the process of biosynthesis and accumulation of starch, which is one of the research are of interest. Like in bacteria, the process of starch biosynthesis in plants occurs by using ADP-Glc as raw materials to made long link chain α-1.4-glucoside. ADP-Glcpyrophosphorylase (AGPase) has important role in biosynthesis of ADP-Glc molecule, and therefore AGPase is an critical enzyme of glycogen synthesis in bacteria and starch in plants. In this study, the gene …show more content…
Beside, cassava is used for many different purposes in industrial starch, such as cosmetics, pharmaceuticals, especially biofuels (Sriroth, Piyachomkwan et al. 2000). Along with economic development, the amount of starch needed for industry is increasing remarkably in developing countries. To meet the starch used, the scientists have applied technical scientific advances mainly transgenic techniques in plant breeding. The synthesis of starch in plants needed 3 enzymes are ADP glucose pyrophosphorylase (AGPase; EC 2.7.7.23), starch synthase (SS; EC 2.4.1.21) and starch branching enzyme (SBE; EC 2.4.1.28) (Fernie, Willmitzer et al. 2002). To improve starch content, we can enhance the activity of three enzymes. The rate of starch synthesis is determined mainly by the activity of AGPase, the first enzyme in the pathway (Zeeman, Kossmann et al. 2010). AGPase catalyzes the first step in the biosynthesis of starch in the plastids by converting Glc1P and ATP into ADPGlc and PPI (Tiessen, Hendriks et al. 2002). The enzyme AGPase has been successfully transfered into a number of crops such as cassava (Ihemere, Arias‐Garzon et al. 2006), potatoes (Ball, Marianne et al. 1991, Stark, Timmerman et al. 1992, Thro 1996), wheat (Smidansky, Clancy et al. 2002), rice (Smidansky, Clancy et al. 2002, Sakulsingharoj, Choi …show more content…
The antibiotic kanamycin, rifamicine, cefotaxime, carbecillin were purchased from Merck.
Synthetized genes encoding AGPase enzyme
Based on the genetic structure of glgC (code V00281.1) encoding AGPase enzymes in E. coli has been published in GenBank and characteristics of the AGPase enzyme in plants, we modified the sequence of glgC gene by creating G336D mutation.
Nucleotide segment encodes for signal peptide chain protein in chloroplasts are derived from the rbcS gene encoding the small section of ribulose 1.5 bisphosphate carboxylase enzyme in Arabidopsis is attached on the 5’ position, and the sequence coding for the tail of c-myc and KDEL is attached on the 3’ position of the DNA segment. Recognized location of XbaI restriction enzyme is attached on5’ position and the recognized location of SacI is added on 3’ position of the gene. AGPase encoding gene (Lan1_opt) is purchased from Integrated DNA Technologies (USA) and is cloned in the vector pUCIDT-AMP.
Design transgenic plant vector carrying pBI121 structure: Lan1_opt Lan1_opt target gene was amplified by transforming the vector pUCAMP: Lan1_opt into E. coli
The miniprep consisted of isolating the DNA plasmid from the bacterial cells. This was used to identify the success of EGFP ligation into pET41a(+) vector upon restriction digest and gel electrophoresis. Additionally, Polymerase Chain Reaction (PCR) was run on the isolated DNA plasmids with one of the primers specifically annealing to a part of pET41a(+) sequence and the other annealing to the EGFP gene.
In the short story “Star Food” by Ethan Canin, Dade is a young teenage boy who works at his father’s grocery store, and from time to time tends to slack off. Dade enjoys relaxing on the roof of the store to observe the clouds, amid dreaming in regards for the life ahead of him, which is what his mother determines is key for him and his future. His father furthermore, believes that Dade should increase the amount of help he provides in the store so that he may one day own the family business. While engaged in his work at the store Dade encounters a woman who steals a loaf of rye bread right before his eyes, but Dade does not do anything; he just stands there, allowing her to shoplift, due to the fact that he has something else on his mind. Dade
The aim of this experiment was to isolate cDNA molecule CIH-1 (Colletotrichum lindemuthianum CIH1 gene) that is contained in vector pBK-CMV and transfer it into cloning vector pUC19. This was attempted by conducting a restriction digest of vectors pUC19 and pBk-CMV containing CIH-1, using restriction endonucleases Xba1 and EcoR1 and the characterization of recombinant plasmids.
In the short story “Star Food” by Ethan Canin, Dade is a sensitive teenager that does not know what direction his life should take. His father is a hardworking man who worries that Dade will not turn out to be like him in his work ethic and overall success. While his mother encourages him to keep being himself and thinks he will change into the person that he is supposed to be. Suddenly Dade is left conflicted when he lets the mysterious women steal from their store, and begins to struggle because of his indecisiveness and self-reflection on his life choices. Ultimately Dade lets the women go because he felt a connection with her struggles.
Modern biotechnology was born at the hands of American scientists Herb Boyer and Stain Cohen, when they developed “recombinant deoxyribonucleotide, (rDNA), [1] for medicinal purposes. Subsequently, biotechnologists started genetically engineering agricultural plants using this technology. A single gene responsible for a certain trait, from one organism (usually a bacterium) is selected altered and then ‘spliced” into the DNA of a plant to create an agricultural crop consisting of that...
Abstract: Enzymes are catalysts therefore we can state that they work to start a reaction or speed it up. The chemical transformed due to the enzyme (catalase) is known as the substrate. In this lab the chemical used was hydrogen peroxide because it can be broken down by catalase. The substrate in this lab would be hydrogen peroxide and the enzymes used will be catalase which is found in both potatoes and liver. This substrate will fill the active sites on the enzyme and the reaction will vary based on the concentration of both and the different factors in the experiment. Students placed either liver or potatoes in test tubes with the substrate and observed them at different temperatures as well as with different concentrations of the substrate. Upon reviewing observations, it can be concluded that liver contains the greater amount of catalase as its rates of reaction were greater than that of the potato.
The Structure and Function of Carbohydrates Large biological molecules are called macromolecules, there are giant molecules (polymers) made up of repeating units (monomers). Carbohydrates are one of the main classes of biological molecules. Macromolecule units (monomers) are joined together by condensation reactions and hydrolysis reactions split macromolecules down into their individual units. Carbohydrates are molecules that contain elements of carbon, hydrogen, and oxygen. Carbohydrates have a 2:1 hydrogen to oxygen ratio, there are twice as many hydrogen atoms as oxygen atoms (the same proportion as in water).
Glycoside Hydrolases are classified into 108 families according with the amino acid sequence similarities. One of these families is GH1 (Glycoside Hydrolases 1), this family consists of enzymes with various substrate specificities, and the enzymes are present is bacteria, Archaea and Eukaryota. The 3D structure of 18 of these enzymes had been determined, and although the extent of sequence varies between 17% and 45%, all the enzymes have a common (β/α)8-barrel motif, and two catalytic glutamate residues located at the C-terminal end of β-strands 4 and 7, which may give a clue about the mechanism of these enzymes.
Investigating the Rate of Reaction between Amylase and Starch. Plan Aim: To be able to The aim of this investigation is to find out whether the volume of amylase affects the rate of reaction between amylase and starch. Prediction: I predict that the greater the volume of amylase then the faster the rate of reaction between the starch and amylase. I predict this because of the lock and key hypothesis.
Investigating the Effect of Enzyme Concentration on the Hydrolysis of Starch with Amylase Aim: Investigate the effect of enzyme concentration on the rate of an enzyme-controlled reaction. Using amylase and starch as my example. Introduction: I am investigating the effect of the concentration of the enzyme, amylase on the time taken for the enzyme to fully breakdown the substrate, starch to a sugar solution. The varied variable will be the concentration and all other variables are going to be fixed. The different concentrations will be: 0.5% 0.75% 1.0% 1.5% 2% An enzyme is a class of protein, which acts as a biological catalyst to speed up the rate of reaction with its substrates.
The term GM foods or GMO (genetically-modified organisms) is most commonly used to refer to crop plants created for human or animal consumption using the latest molecular biology techniques (Whitman, 2000). These plants have been modified in the laboratory to offer desired traits such as increased resistance to herbicides or improved nutritional content. Also, genetic engineering techniques have been applied to create plants with the exact desired trait very rapidly and accurately. For example, this is done by the geneticist isolating the gene responsible for drought tolerance and inserts it into another plant. The new genetically-modified plant will now have gained drought tolerance as well.
Wheat is one of the major food crops in the world, which provides 532 kcal/capita/day (http://faostat.fao.org). World production of wheat is about 670 M tonnes, which is only behind maize (872 M tonnes) and rice (720 M tonnes) (http://faostat.fao.org) whereas the US is the third largest wheat producer in the world (http://faostat.fao.org). Wheat group comprise of 13 diploid and 18 allopolyploid species (12 tetra and 6 hexa) (Feldman et al. 2012), where hexaploid wheat is the most cultivated class throughout the world (Faris 2014). Hexaploid wheat (bread wheat) is known as the allohexaploid (2n=6x=42, BBAADD), because it is derived from the diploid species via convergent evolution (Faris 2014 and Feldman et al. 2012). Hexaploid wheat consist of winter and spring wheat, which mainly differed by vernalization governed by vrn (vrn1, vrn2) genes (Taiz and Zeiger 2002, Doebley et al. 2006).
Genetic engineering enhances plant resistance to drought, salinity, disease, pests and herbicides. The aim is to try and enhance the growth, productivity, nutrient value, and chemical composition of the plants. Chemicals are constantly being developed or improved to enhance the competitiveness and adaptability of crops, and to kill the parasites and weeds which plague the agricultural sector. . This however is not always good as the plant and the pests then become resistant to these new chemicals defeating the purpose of it being used. The new chemicals which are produced to kill these strong pests and weeds may be more harmful to other plants and remove nutrients within the soil in turn reducing the yield of agricultural crops. The benefits of these characteristics are seen in Argentina according to Pelletier (2010) as they use glyphosphate resistant soybean which allowed the comeback of this crop, as the so...
1443 - 1460. Keener, K., Hoban, T. and Balasubramanian, R. 2014. Biotechnology and its applications. [online] Available at: http://www.ces.ncsu.edu/depts/foodsci/ext/pubs/bioapp.html [Accessed: 11 Apr 2014].
As they always say HEALTH IS WEALTH! Your food selection today, makes you enjoy for the moment and affects your health tomorrow and in the future!