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Scale culture and promotion technique for trans TNF-alpha gene blue algae growth and gene expression by light reactor

A gene expression and photoreactor technology, which is applied in the field of photoreactor large-scale cultivation and promoting the growth and gene expression of TNF-α gene cyanobacteria

Inactive Publication Date: 2008-09-17
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Separation and harvesting of algae cells and crushing

Method used

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  • Scale culture and promotion technique for trans TNF-alpha gene blue algae growth and gene expression by light reactor
  • Scale culture and promotion technique for trans TNF-alpha gene blue algae growth and gene expression by light reactor
  • Scale culture and promotion technique for trans TNF-alpha gene blue algae growth and gene expression by light reactor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0023] A closed 100L photoreactor was established to cultivate cyanobacteria. The main reactor is cylindrical, with a height of 110 cm, a diameter of 35 cm, and a usable volume of 70 L. There are three built-in lamp tubes, and each lamp tube has 6 built-in lamp tubes, and the power of each lamp tube is 18w. Illumination area 0.3617m 2 Unit illumination area 4.4589m -1 . The ventilation volume is 10L / min. The culture conditions are as follows: both the transgenic type 7120 and the wild type 7120 are cultured in BG-11 medium. BG-11 medium was sterilized at 121°C for 20 minutes. The initial pH value is 8.0, and the culture temperature is 25°C. The wild-type 7120 culture period was 12 days, and the transgenic type culture period was 15 days. Take samples every two days to measure OD 750 . Wild type 7120OD when harvesting algae 750 was 0.428; transgenic type 7120OD 750 is 0.524. See figure 2 . After 12 hours of precipitation, the supernatant was discarded, and the al...

Embodiment example 2

[0025] Improve the closed 100L photoreactor to cultivate cyanobacteria. The main reactor is cylindrical, with a height of 110 cm, a diameter of 35 cm, and a usable volume of 70 L. There are three built-in lamp tubes, and each lamp tube has 6 built-in lamp tubes, and the power of each lamp tube is 18w. Illumination area 0.3617m 2 Unit illumination area 4.4589m -1 . add CO 2 , CO 2 The amount added is controlled at 5% with a flow meter. The ventilation volume is 10L / min. Both the transgenic type 7120 and the wild type 7120 were cultured in BG-11 medium. BG-11 medium was sterilized at 121°C for 20 minutes. The initial pH value is 8.0, and the culture temperature is 28°C. The wild-type 7120 culture period is 12 days, and the transgenic type culture period is 12 days. Daily sampling for OD 750 . Wild type 7120 OD when harvesting algae 750 was 0.545; transgenic type 7120 OD 750 is 0.563. See image 3 . After 12 hours of precipitation, the supernatant was discarded, ...

Embodiment example 3

[0027] Then improve the closed 100L photoreactor to cultivate cyanobacteria. The main reactor is cylindrical, with a height of 110 cm, a diameter of 35 cm, and a usable volume of 70 L. There are three built-in lamp tubes, and each lamp tube has 6 built-in lamp tubes, and the power of each lamp tube is 18w. Illumination area 0.7571m 2 Unit illumination area 9.335m -1 . Both the transgenic type 7120 and the wild type 7120 were cultured in BG-11 medium. BG-11 medium was sterilized at 121°C for 20 minutes. After sterilization, a glucose solution sterilized at 115° C. for 20 minutes was added to the reactor to make the glucose concentration in the reactor 50 mmol / L. The ventilation volume is 10L / min. The initial pH value is 8.0, and the culture temperature is 28°C. The wild-type 7120 culture period was 8 days, and the transgenic type culture period was 8 days. Daily sampling for OD 750 . Wild type 7120 OD when harvesting algae 750 was 0.549; transgenic type 7120 OD 750 ...

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Abstract

The invention relates to a 100-liter close type photo-bioreactor and a processing technique for the mass cultivation and promotion of the growth and the gene expression of trans-TNF-Alfa anabaena by using the photo-bioreactor. The reactor comprises three built-in light transmitting glass tube, an adjustable illumination facility, a sterile air blowing facility, a high temperature sterilizing system, and CO2 distribution facility. A culture medium is prepared, and regulated and controlled artificially in the reactor so as to promote the growth and the gene expression of the trans-TNF-Alfa anabaena. The production method is suitable for large scale production of transgene blue algae, and has the advantages of high stability, strong controllability and maneuverability, small outcome difference between batches, etc.

Description

technical field [0001] The invention relates to a closed photoreactor for large-scale cultivation and a production process for promoting the growth and gene expression of TNF-a-transformed cyanobacteria. Background technique: [0002] Human tumor necrosis factor-α (human tumor necrosis factor-α, hTNF-α) is a non-glycosylated protein produced by activated mononuclear macrophages, which has a specific killing effect on tumor tissue and tumor cells, and is expected to be developed become a promising antitumor drug. However, its use is limited due to its severe clinical side effects. In this regard, we plan to use cyanobacteria genetic engineering to prepare oral drugs to alleviate toxic and side effects. [0003] Cyanobacteria, also known as cyanobacteria, are a group of prokaryotic organisms with various species and wide distribution that can perform oxygen-evolving photosynthesis. In recent years, the development of molecular biology of cyanobacteria has made cyanobacteria...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/28C12N1/13C12M1/36
CPCC12M21/02C12M31/10C12M41/12C12M29/26
Inventor 施定基朱学义张宏业李轩左维明张明华章华西李爽李丹张芃芃童艳宋东辉
Owner TIANJIN UNIV OF SCI & TECH
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