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Blending culture for sponge and intergrowth microbe, culturing colony monitoring method

A technology of mixed culture and sponge symbiosis, which is applied in the measurement/testing of microorganisms, biochemical equipment and methods, microorganisms, etc., and can solve problems that are difficult to achieve

Inactive Publication Date: 2006-05-03
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

[0007] For the dynamic monitoring of microbial populations in the process of mixed microbial fermentation culture, the current conventional technology is still difficult to achieve
16S rDNA-DGGE (denaturing gradient gel electrophoresis) gene fingerprint technology has become a powerful tool to reveal the structure of complex microbial flora, and is also very suitable for the dynamic detection of microbial population structure over time in the process of mixed microbial culture. Any report on the application of technology to monitoring the population of sponge symbiotic microorganisms in the mixed culture process

Method used

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  • Blending culture for sponge and intergrowth microbe, culturing colony monitoring method
  • Blending culture for sponge and intergrowth microbe, culturing colony monitoring method

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Embodiment Construction

[0017] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0018] Realize the technical route of the inventive method as figure 1 As shown, first prepare the mixed bacteria seed solution, prepare the sponge extract juice, and prepare the compound medium containing the sponge extract juice; inoculate the mixed bacteria seed solution, and carry out the mixed culture of the sponge symbiotic microorganisms; sample and extract the total DNA of the mixed cultured microorganisms during the culture , 16S rDNA was amplified by PCR, and a gene fingerprint map reflecting the structure of the mixed cultured microbial population was constructed by DGGE; the bacteria represented by each band were analyzed through rubber tapping recovery of fingerprint bands, cloning sequencing and homology and phylogenetic analysis Molecular characterization enabling monitoring of commensal microbial populations in...

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Abstract

The mixed cultivation and culturing population monitor method for sponge symbiotic microbe comprise: cleaning the epiphytic microbe on sponge surface, dipping into artificial sea water to shock and release symbiotic microbe in sponge for preparing germ liquid; extracting and preparing sponge draw-off juice with alcohol to add into seawater culture medium and prepare composite medium; inoculating to culture mixed microbe; extracting the whole DNA of cultured microbe to amplify into 16S rDNA segment less than 500bp by PCR for modified gradient gel electrophoresis and obtaining gene fingerprint pattern; realizing in-vitro monitor by callbacking fingerprint pattern strips of different time, cloning, sequencing, and homological and phylogenetic analysis to identify germ kind. This invention can improve separation and culture efficiency, and provides opposite material and technique support.

Description

technical field [0001] The invention relates to a method for monitoring the mixed culture of sponge symbiotic microorganisms and cultivable populations, in particular to the monitoring of the mixed culture of sponge symbiotic microorganisms and the population structure of cultivable microorganisms in the presence of sponge extracts. It belongs to the technical field of marine microbial engineering. Background technique [0002] Sponges are one of the marine organisms with the most abundant marine active substances, but due to source limitations, few active substances have entered the clinical research stage. At present, more and more studies have shown that some active substances with medicinal value are produced by sponge symbiotic microorganisms. Therefore, using microbial fermentation engineering technology may be the most feasible way to prepare sponge active substances in large quantities. [0003] The traditional microbial fermentation culture is based on pure bacter...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12Q1/68
Inventor 李志勇何丽明
Owner SHANGHAI JIAO TONG UNIV
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