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Method for screening sponge-associated antibacterial active strains

A technology of co-adherence and antibacterial activity of sponges, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of affecting active substances, blind research on the characteristics of active substances, etc., and achieve the effect of enhancing the purpose.

Inactive Publication Date: 2010-10-27
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

This method has the following shortcomings: gene silencing, screening conditions, etc. affect the detection results of active substances, and there is a lot of blindness in the research on the characteristics of active substances

Method used

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  • Method for screening sponge-associated antibacterial active strains
  • Method for screening sponge-associated antibacterial active strains
  • Method for screening sponge-associated antibacterial active strains

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Embodiment Construction

[0020] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments. The following examples are not intended to limit the present invention.

[0021] The flow process of the inventive method is as figure 1 As shown, firstly, isolate and purify the sponge symbiotic bacteria, then extract the sponge symbiotic bacteria DNA, use the sponge symbiotic bacteria DNA as a template, design primers for the polymerase chain reaction of the ks domain of the bacterial pks gene fragment, The 690bp fragment of the sponge symbiotic bacteria obtained by screening was sequenced and analyzed, and then the antibacterial activity of the pks positive strain was determined, and finally the antibacterial activity of the sponge symbiotic bacteria was identified in molecular biology to determine the pks gene fragment positive sponge Species of symbiotic antibacterially active strains.

[0022] In the embodiment of the pr...

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Abstract

The invention relates to a method for screening sponge-associated antibacterial active strains, combining marine bacteria pks gene molecular biology screening with antibacterial activity detection. The method comprises the following steps of: first, separating and purifying sponge-associated strains, extracting sponge-associated strain DNA as templates by a phenol-chloroform method; taking a ks region universal primer GB1 / GB2 of bacterial pks gene as a primer to perform polymerase chain reaction for amplifying the pks gene fragment; obtaining a 690bp fragment of sponge-associated bacterial, sequencing, then submitting to Gengank and carrying out Blast analysis; measuring the antibacterial activity of pks positive strains; taking screened antibacterial active strains having 690bp target fragments as templates; taking 16SrDNA universal primer 27f / 1492r as a primer; carrying out polymerase chain reaction to amplify the target fragments and sequencing identification; submitting obtained sequence to Genbank; and carrying out Blast analysis to determine the species of pks gene fragment positive sponge-associated antibacterial active strains.

Description

technical field [0001] The present invention relates to a method for screening antibacterial active bacterial strains of sponge symbiosis, in particular to a method for screening antibacterial active bacterial strains of symbiotic sponges which combines polyketide synthase functional gene screening and antibacterial activity, and belongs to the field of microbial molecular technology . Background technique [0002] Sponges are a type of multicellular animal with the simplest structure and the most primitive form, and are an important source of marine natural medicines. The sources of sponge active substances are complex, most of which come from the symbiotic microorganisms of sponges. Due to the special growth environment, sponge symbiotic microorganisms can produce metabolites with novel structures and unique activities, which are important marine medicinal microbial resources. Polyketides are a class of natural products with complex structures and wide uses catalyzed by ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 张薇张风丽李志勇
Owner SHANGHAI JIAO TONG UNIV
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