Difference between revisions of "Part:BBa K3852003"

Revision as of 12:59, 17 October 2021

PHO3

Introduction

The expression of PHO3 gene in S. cerevisiae can hydrolyze thiamine phosphate in the pericytoplasmic space and increase thiamine uptake by cells.

Experiment

In our experiments, we found that overexpression of this gene could promote the tolerance of S. cerevisiae to high temperature. However, if the samples we tested were hot soup or other food, s. cerevisiae used for testing might lose activity. If the samples were tested after cooling, it would not only take too long, but also might change the flavor of the samples. Therefore, we intend to overexpress PHO3 gene in our strain to improve its heat resistance and enable it to detect a wider range of samples. In the following sections, we will explain how the PHO3 overexpression can improve the heat resistance of S. cerevisiae.

Comparison of yeast growth status

Our experiment was divided into two groups, one group was PHO3 overexpressed strain (experimental group), the other group was normal strain (control group), using the same conditions at 37. C, culture in 20g/L glucose solution for 36h. The measured data are drawn as the following figure. It was not difficult to see that the growth status of PHO3 overexpressed strain was obviously better than that of the normal strain, which indicated that the overexpression of PHO3 gene had a high-temperature resistance effect on S. cerevisiae. This gene will subsequently be overexpressed in our saccharomyces cerevisiae strains to make them resistant to high temperatures.

Figure1. Saccharomyces cerevisiae growth curve

Reference

[1]1To-E A, et al. (1973) Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae. J Bacteriol 113(2):727-38 [2] Doi S, et al. (1989) Induction of repressible acid phosphatase by unsaturated fatty acid in Saccharomyces cerevisiae. J Cell Sci 94 ( Pt 3):511-6 [3] Praetorius-Ibba M, et al. (1997) Homologous recombination partly restores the secretion defect of underglycosylated acid phosphatase in yeast. Curr Genet 32(3):190-6 [4] Kalebina TS, et al. (2008) The role of high-molecular-weight polyphosphates in activation of glucan transferase Bgl2p from Saccharomyces cerevisiae cell wall. Dokl Biochem Biophys 420:142-5

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 610
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K3852003 short</partinfo>
-This gene encodes a member of a family of candidate taste receptors that are members of the G protein-coupled receptor superfamily and that are specifically expressed by taste receptor cells of the tongue and palate epithelia. These apparently intronless genes encode a 7-transmembrane receptor protein, functioning as a bitter taste receptor. This gene is clustered with another 3 candidate taste receptor genes in chromosome 7 and is genetically linked to loci that influence bitter perception.
+===Introduction===
-Several papers have shown that bitter substances are recognized by the bitter receptor TAS2R, a family of G protein-coupled receptors expressed in taste tissue. In humans, more than 25 TAS2Rs have been identified; some of these are activated by bitter substances and function as bitter taste receptors. Because many bitter tastants activate hTAS2R, it has been thought that humans perceive bitterness via those receptors. Experiments show that a variety of bitter peptides can activate T2R4.In our experiments, we used this gene to express bitter taste receptors in Saccharomyces cerevisiae.We acquire it from synthesis company.
+The expression of PHO3 gene in S. cerevisiae can hydrolyze thiamine phosphate in the pericytoplasmic space and increase thiamine uptake by cells.
-===Usage and Biology===
+===Experiment===
-Gustducin-coupled receptor for denatonium and N6-propyl-2-thiouracil implicated in the perception of bitter compounds in the oral cavity and the gastrointestinal tract. Signals through PLCB2 and the calcium-regulated cation channel TRPM5. In airway epithelial cells, binding of denatonium increases the intracellular calcium ion concentration and stimulates ciliary beat frequency.
+In our experiments, we found that overexpression of this gene could promote the tolerance of S. cerevisiae to high temperature. However, if the samples we tested were hot soup or other food, s. cerevisiae used for testing might lose activity. If the samples were tested after cooling, it would not only take too long, but also might change the flavor of the samples. Therefore, we intend to overexpress PHO3 gene in our strain to improve its heat resistance and enable it to detect a wider range of samples. In the following sections, we will explain how the PHO3 overexpression can improve the heat resistance of S. cerevisiae.
+===Comparison of yeast growth status===
+Our experiment was divided into two groups, one group was PHO3 overexpressed strain (experimental group), the other group was normal strain (control group), using the same conditions at 37. C, culture in 20g/L glucose solution for 36h. The measured data are drawn as the following figure. It was not difficult to see that the growth status of PHO3 overexpressed strain was obviously better than that of the normal strain, which indicated that the overexpression of PHO3 gene had a high-temperature resistance effect on S. cerevisiae. This gene will subsequently be overexpressed in our saccharomyces cerevisiae strains to make them resistant to high temperatures.
+[[File:T--BIT-China--Engeering 15.png|550px|thumb|center|Figure1. Saccharomyces cerevisiae growth curve]]
 ===Reference===
-[1] Mueller, K., Hoon, M., Erlenbach, I. et al. Erratum: The receptors and coding logic for bitter taste. [J]Nature 446, 342 (2007).
+[1]1To-E A, et al. (1973) Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae. J Bacteriol 113(2):727-38
+[2] Doi S, et al. (1989) Induction of repressible acid phosphatase by unsaturated fatty acid in Saccharomyces cerevisiae. J Cell Sci 94 ( Pt 3):511-6
+[3] Praetorius-Ibba M, et al. (1997) Homologous recombination partly restores the secretion defect of underglycosylated acid phosphatase in yeast. Curr Genet 32(3):190-6
+[4] Kalebina TS, et al. (2008) The role of high-molecular-weight polyphosphates in activation of glucan transferase Bgl2p from Saccharomyces cerevisiae cell wall. Dokl Biochem Biophys 420:142-5
-[2] Kenji Maehashi, Mami Matano, Hong Wang et al.Bitter peptides activate hTAS2Rs, the human bitter receptors[J]Biochemical and Biophysical Research Communications.2008,365(4):
--855.