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Semi-synthetic culture medium for dictyostelium discoideum and preparing method thereof

A semi-synthetic and culture medium technology, applied in the direction of fungi, etc., can solve the problems of slow cell growth and achieve the effects of increasing production, increasing gene expression, and reducing costs

Inactive Publication Date: 2007-08-15
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the use of synthetic media can usually achieve higher cell density, the growth rate of cells in synthetic media is relatively slow, and the generation time is usually 14-16h

Method used

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  • Semi-synthetic culture medium for dictyostelium discoideum and preparing method thereof
  • Semi-synthetic culture medium for dictyostelium discoideum and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: the selection of carbon source type

[0046] On the basis of the initial culture medium, glucose was replaced with glucose, maltose, mannose, sucrose, fructose, lactose and glycerol at the same concentration (both 10.0 g / L) to prepare different culture media. Shake flask cultures of Dictyostelium discoideum were carried out under the same culture conditions (as described in the instructions). The initial medium was HL-5C medium. For comparison, the cultivation of Dictyostelium discoideum in the complex medium without adding carbon source was also carried out. The result is shown in Figure 1. The results showed that when glucose or maltose was used as the sole carbon source, the cells grew well, and the maximum cell density could reach 2.0×10 7cells / mL, and the cells grow rapidly. Both lactose and sucrose inhibited cell growth, glycerol had little effect on its growth, while fructose and mannose both promoted cell growth, but their effect was not as goo...

Embodiment 2

[0047] Embodiment 2: Optimization of glucose concentration

[0048] On the basis of the initial medium, different concentrations of glucose (0g / L, 5g / L, 10g / L, 15g / L, 20g / L, 30g / L) were used as carbon sources to prepare medium. Shake flask cultures of Dictyostelium discoideum were carried out under the same culture conditions (as described in the instructions). The initial medium was HL-5C medium. The result is shown in Figure 2. The results show that the glucose concentration directly affects the maximum cell density, and the optimal glucose concentration is in the range of 10-15g / L. At this time, Dictyostelium grows well, and the maximum cell density can reach 1.9×10 7 cells / mL. When glucose is insufficient, the cell density will decrease significantly, only reaching 2.6×10 6 cells / mL. But when the glucose concentration was higher than 20g / L, the maximum cell density decreased instead.

Embodiment 3

[0049] Embodiment 3: Optimization of maltose concentration

[0050] On the basis of the initial medium, glucose was replaced with maltose of different concentrations (0g / L, 5g / L, 10g / L, 15g / L, 20g / L, 25g / L, 30g / L) to prepare the medium. Shake flask culture of Dictyostelium discoideum was carried out under the same culture conditions. The initial medium was HL-5C medium. The result is shown in Figure 3. The results show that the concentration of maltose directly affects the maximum cell density, and the optimum concentration of maltose is in the range of 15-20 g / L. At this time, Dictyostelium grows well, and the maximum cell density can reach 1.9×10 7 cells / mL. When the medium lacked maltose, the maximum cell density was greatly reduced and only reached 2×10 6 cells / mL. When the initial maltose concentration is higher than 20g / L, the cell growth is inhibited instead, and the maximum cell density is low. For example, when the initial maltose concentration is as high as 30g / ...

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Abstract

The invention discloses a preparing method of semi-synthetic medium of dish group net prosthecae, which comprises the following steps: choosing yeast powder, bacteriological tryptone, casein peptone, protein peptone and carbon source; setting amylaceum or barley sugar as carbon source; choosing KCl, MgCl2 and CaCl2 as inorganic salt; choosing KH2PO4 and Na2HPO4 as microcosmic salt cushioning liquid; allocating the other component as water; adding yeast powder, bacteriological tryptone, casein peptone, protein peptone into inorganic salt and microcosmic salt cushioning liquid; adjusting pH value with NaOH or H3PO4; sterilizing with high pressure; getting semi-synthetic medium without carbon source; allocating amylaceum or barley sugar solution; sterilizing with high pressure or degerming with film filtration; adding in to semi-synthetic medium without carbon source.

Description

technical field [0001] The invention relates to a bacterial culture medium, in particular to a semi-synthetic culture medium suitable for high-density cultivation of Dictyostelium discoideum and production of genetic engineering products. Background technique [0002] Currently, the commonly used expression system hosts include prokaryotic Escherichia coli, eukaryotic yeast, insect cells, and animal cell lines. With the deepening of recombinant protein drug research and the research requirements of the functional genomics era, the above-mentioned commonly used systems have limitations in varying degrees in high-throughput expression of eukaryotic genes. The E. coli expression system usually does not have various post-translational modification processing functions, and the expressed heterologous protein usually exists in the cell in the form of insoluble inclusion bodies, which is not suitable for large-scale production of recombinant heterologous proteins. Yeast can hyperg...

Claims

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Application Information

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IPC IPC(8): C12N1/14
Inventor 卢英华王颖何宁徐志南陈翠雪凌雪萍
Owner XIAMEN UNIV
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