CN103882005A - Mutation screening method and application of streptomyces strain SPC6-50-1 - Google Patents
Mutation screening method and application of streptomyces strain SPC6-50-1 Download PDFInfo
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- CN103882005A CN103882005A CN201410057958.4A CN201410057958A CN103882005A CN 103882005 A CN103882005 A CN 103882005A CN 201410057958 A CN201410057958 A CN 201410057958A CN 103882005 A CN103882005 A CN 103882005A
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Abstract
The invention discloses a mutation screening method of a streptomyces strain SPC6-50-1. The mutation screening method comprises the steps of inducing mutation of streptomyces coelicolor and streptomyces Lzsg6 through nitrosoguanidine to obtain a streptomyces mutant library, and screening the mutant library through rifampicin and streptomycin to obtain a strain with a transcription system and a translation system in mutation. The invention also discloses a streptomyces strain SPC6-50-1 and application thereof. The streptomyces strain SPC6-50-1 disclosed by the invention has the beneficial effects that the strain contains substances having bacteriostatic activities, and the method of nitrosoguanidine (NTG) inducing as well as rifampicin and streptomycin screening can activate silent genes of the streptomyces to obtain a new metabolite, thus obtaining a new antibiotic; meanwhile, the mutation screening method has the advantages of simple operation, short cycle and high efficiency, and is not only applicable to streptomyces but also suitable for such microorganisms as myxobacteria rich in secondary metabolites; the mutation screening method has good popularization and application values.
Description
Technical field
The present invention relates to production of antibiotics field, be specifically related to mutagenesis screening method, streptomycete bacterial strain SPC6-50-1 and the application thereof of streptomycete bacterial strain SPC6-50-1.
Background technology
Since the nineteen twenty-eight mankind find microbiotic first---since penicillin, in these decades, microbiotic industry develop rapidly, for the mankind healthy worked it out huge contribution.But due to people irrational use microbiotic, especially China, the problem of antibiotic resistance breaks out out gradually.Address this problem, except noting antibiotic reasonable use, develop that new resistance germ is had to the active new antibiotic of inhibition is also very important.
Streptomycete is that one is extensively present in nature, especially in soil, and protokaryon, gram-positive microorganism, belongs to actinomycetes door.Streptomycete has complicated Morphological Differentiation and secondary metabolism, and can produce the multiple bioactive secondary metabolite that has in the transition period of nourishing and growing and spore breaks up.The metabolite that this genus produces, except microbiotic, also comprises antineoplastic agent, immunosuppressor, pest-resistant dose and the outer lytic enzyme of born of the same parents, as amylase, chitinase cellulase, polygalacturonase, zytase and proteolytic enzyme etc.These secondary metabolites all have important application in fields such as medical treatment, agricultural, food.
That finds so far the forties in 20th century more than 12000 plants in microbe-derived biologically active substance, more than 55% is produced by streptomycete, approximately has 70% to be produced by streptomycete in the microbiotic of having found.The microbiotic approximately 2/3rds of application derives from streptomyces clinically at present.
Sum up the microbiotic quantity obtaining in the past few decades from streptomycete, since finding the nineties, the quantity that it is found that new antibiotic sharply reduces, and traditional the passing through of this explanation screened the method that new streptomycete obtains new antibiotic and run into bottleneck (Berdy, 2005).Find a kind of new efficient acquisition new antibiotic method necessary.
At present, (Streptomyces grise μ several streptomycetes such as s) has completed genome sequencing for streptomyces coelicolor (Streptomyces coelicolor), Avid kyowamycin (Streptomyces avermitilis), streptomyces griseus.Researchist's discovery, in the gene of streptomycete linear structure, the main cell function important to some of the central core district gene of high conservative is relevant, and has a large amount of secondary metabolite gene clusters (Chen et al., 2002 in both sides, core area; Ohnishi et al., 2008).For example: at the genome of analyzing streptomyces coelicolor, find that there is 23 gene clusters that participate in forming complicated secondary metabolite, account for the 5%(Bentley et al. of genome total amount, 2002), as shown in table 1 is that Streptomyces coelicolor A3 (2) produces secondary metabolite gene cluster; In Avid kyowamycin, there are 30 secondary metabolism gene clusters, account for complete genomic 6.6%(Ikeda et al., 2003).Therefore,, from this angle, whole streptomyces is exactly a huge potential secondary metabolite storehouse.Make streptomycete transcribe, translate these genes by some method, and to filter out the secondary metabolite with active substance be a kind of effective way (Zazopo μ los et al., 2003) that obtains new antibiotic.
Table 1
In streptomycete, coding secondary metabolism particularly gene and the approach specificity regulatory protein gene Chang Liansuo of microbiotic biosynthetic pathway exists.In gene cluster, often comprise one to multiple approach specificity regulatory genes, they regulate and control one or more and have the antibiotic biosynthesizing of common biosynthetic pathway.As: the genetic expression of the actinorhodin that streptomyces coelicolor produces and undecyl bacterium spirit red pigment is by its special transcription activating protein ActII-ORF4 and RedD regulation and control (van Wezel and McDowall, 2011).In streptomycete, most secondary metabolites synthetic genes and specificity regulatory gene are all reticent, only in running into some particular surroundings, corresponding special pathways metabolism regulatory gene can be activated, produce new meta-bolites or the adjusting to its Physiology and biochemistry, help streptomycete to conform, pull through their difficulties (Challis and Hopwood, 2003).Finding after the biological synthesis gene cluster of certain potential compound, by the method for Combinatorial biosynthesis (Combinatorial Biosynthesis), as this gene cluster is carried out to heterogenous expression; Or cut out and modify or utilize the method for sudden change to change heredity metabolize process with enzyme, just likely obtain new microbiotic (Medema et al., 2011; Okamoto et al., 2003).The people such as Yu Zhibin use microbiotic to obtain 1 strain chlorampenicol resistant and the streptomycin resistance strain of anti-tumor activity to deriving from the non-activity streptomycete B3054 screening of ocean.And through active result analysis, from 1 strain streptomycin resistance strain fermented product, separate and obtain 5 active compound for anti tumor, one of them is new natural product (Yu Zhibin, Zhu Tianjiao, Cui Chengbin etc., 2006).Therefore, obtaining new antibiotic by change streptomycete heredity metabolize approach is a kind of very effective way.
NTG is nitrosoguanidine, is a kind of very strong chemical mutagen.The sudden change that NTG brings out is mainly GC-AT conversion, also has in addition excision among a small circle, phase shift mutation and the right disappearance of GC.First the present invention carries out mutagenesis by superpower mutagenic compound NTG to streptomyces gene group, and the induced mutation rate of NTG can reach tens percent, and is random, so just can obtain a more comprehensive streptomycete mutant library of ratio.And then utilize Rifampin and Streptomycin sulphate to screen.Rifampin can be combined with the β subunit of the RNA polymerase that relies on DNA, and anti-bacteria RNA's is synthetic, prevents that this enzyme is connected with DNA, thereby blocking-up rna transcription process stops the synthetic of DNA and albumen.Screen by Rifampin, can obtain efficiently because of the re-reading system bacterial strain that can grow on the flat board that contains Rifampin of undergoing mutation.These sudden changes have changed original re-reading system, and the sudden change of re-reading system may make the interior regulation and control disorder of bacterium, therefore can make some silencers be expressed, thereby have an opportunity to obtain new antibiotic.Streptomycin sulphate is a kind of aminoglycoside medicine, passes through bacterial cell membrane through active transport, with the specific receptor protein binding of bacterial ribosome 30S subunit, and the formation of initiation complex between interfere information Yeast Nucleic Acid and 30S subunit, arrestin is synthetic.Screening by Streptomycin sulphate can obtain the bacterial strain that translation system is undergone mutation, and these bacterial strains there will be change to a certain degree for protein translation ability.
Summary of the invention
Object of the present invention is exactly for above-mentioned defect of the prior art, mutagenesis screening method, the streptomycete bacterial strain SPC6-50-1 of streptomycete (Streptomyces linzensis) bacterial strain SPC6-50-1 are provided, and this bacterial strain is that the method that efficiently activates the reticent antibiotic resistance gene metabolic gene of streptomycete bunch obtains.
To achieve these goals, technical scheme provided by the invention is: the mutagenesis screening method of streptomycete (Streptomyces linzensis) bacterial strain SPC6-50-1,, by nitrosoguanidine (NTG), streptomyces coelicolor (Streptomyces coelicolor A3 (2)) and streptomycete (Streptomyces sp.) Lzsg6 are carried out to mutagenesis, obtain streptomycete mutant library, by Rifampin and Streptomycin sulphate screening, obtain the bacterial strain that re-reading system and translation system are undergone mutation again.
Further, the mutagenesis screening method of above-mentioned streptomycete (Streptomyces linzensis) bacterial strain SPC6-50-1, includes following steps:
1) chemical mutagen NTG carries out mutagenesis to streptomycete: streptomyces coelicolor (Streptomyces coelicolor A3 (2)) and streptomycete (Streptomyces sp.) Lzsg6 are prepared as to bacteria suspension, with NTG mother liquor, bacteria suspension is carried out to mutagenesis, obtain the bacteria suspension that NTG processed;
2) bacterial strain that selects re-reading system to undergo mutation in order to good fortune plansifter: the bacteria suspension that the NTG that step 1) is obtained processed is evenly applied on the Gause I culture medium flat plate that contains 5 μ g/ml Rifampins, cultivate 7-15d for 30 DEG C, the single bacterium colony growing is connected to respectively in the test tube that 5mL liquid nutrient medium YEME or TSBY are housed, and then be prepared into mutagenesis bacteria suspension, carry out NTG mutagenesis, the bacteria suspension of processing is evenly applied on the Gause I culture medium flat plate that contains 10 μ g/ml Rifampins, repeatedly, carry out 15 μ g/ml, 20 μ g/ml, 25 μ g/ml, 30 μ g/ml, 35 μ g/ml, 40 μ g/ml, 45 μ g/ml, the Rifampin screening of 50 μ g/ml, obtain the rear bacterial strain of Rifampin screening,
3) bacterial strain of undergoing mutation with Streptomycin sulphate screening translation system: by step 2) bacterial strain carries out NTG mutagenesis again after the Rifampin screening that obtains, method and step 2) identical, the bacteria suspension of processing is evenly applied to and contains 5 μ g/mL Streptomycin sulphates, on the Gause I culture medium flat plate of 50 μ g/mL Rifampins, cultivate 7-15d for 30 DEG C, the single bacterium colony growing is connected to respectively in the test tube that 5mL liquid nutrient medium YEME or TSBY are housed, and then be prepared into mutagenesis bacteria suspension, carry out NTG mutagenesis, the bacteria suspension of processing is evenly applied to and contains 10 μ g/mL Streptomycin sulphates, on the Gause I culture medium flat plate of 50 μ g/mL Rifampins, repeatedly, Concentration of Rifampicin is 50 μ g/mL, Streptomycin sulphate is 15 μ g/ml, 20 μ g/ml, 30 μ g/ml, 40 μ g/ml, 50 μ g/ml, 60 μ g/ml, 70 μ g/ml, 80 μ g/ml, 90 μ g/ml, 100 μ g/ml increase progressively successively, obtain Streptomycin sulphate, bacterial strain after Rifampin screening.
Second object of the present invention has been to provide streptomycete (Streptomyces linzensis) the bacterial strain SPC6-50-1 that a strain obtains with above-mentioned mutagenesis screening method mutagenesis screening, described streptomycete (Streptomyces linzensis) SPC6-50-1, be preserved in (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms's common micro-organisms center on October 18th, 2013, Institute of Microorganism, Academia Sinica), the deposit number of streptomycete SPC6-50-1 is CGMCC No.8372.This bacterial strain is Gram-positive, can not forms aerial mycelium, vegetative mycelium for yellow, cultivates one week colony diameter 7mm left and right for 37 DEG C.
The 3rd object of the present invention has been to provide above-mentioned streptomycete (Streptomyces linzensis) bacterial strain SPC6-50-1 in the application of preparing in microbiotic.
The 4th object of the present invention has been to provide the mutagenesis screening method of above-mentioned streptomycete (Streptomyces linzensis) bacterial strain SPC6-50-1 in the application activating in streptomycete silencer.
The 5th object of the present invention has been to provide the mutagenesis screening method of above-mentioned streptomycete (Streptomyces linzensis) bacterial strain SPC6-50-1 in the application of preparing in microbiotic.
Further, above-mentioned application, in the flat lining out of Gause I by the bacterial strain after Streptomycin sulphate, Rifampin screening, collect Conidia preservation, be connected in the triangular flask that liquid nutrient medium YEME or TSBY are housed, at 30 DEG C, on the shaking table of 200r/min, cultivate 3-7d, the centrifugal 1min of 12000r/min, gets supernatant.
Method of the present invention is carried out mutagenesis by NTG to streptomyces coelicolor (Streptomyces coelicolor A3 (2)) and streptomycete (Streptomyces sp.) Lzsg6, obtain a more comprehensive streptomycete mutant library of ratio, then filter out by Rifampin and Streptomycin sulphate the bacterial strain that re-reading system and translation system are undergone mutation, the fermented liquid of these bacterial strains is carried out to bacteriostatic activity detection, find to change before its metabolite activity, this explanation this method can activate some silencers of streptomycete, obtain new meta-bolites, thereby obtain new antibiotic.
Beneficial effect of the present invention is: streptomycete provided by the invention (Streptomyces linzensis) bacterial strain SPC6-50-1 has the material of bacteriostatic activity, and the method for its NTG induction, Rifampin and Streptomycin sulphate screening can activate some silencers of streptomycete, obtain new meta-bolites, obtain new antibiotic, simultaneously, above-mentioned induction screening method has simple to operate, cycle is short, the advantage that efficiency is high, not only applicable to streptomycete, also being applicable to slime bacteria etc. has the microorganism of enriching secondary metabolite, has good value for applications.
Brief description of the drawings
Fig. 1 is the concrete technology schema of the mutagenesis screening method of streptomycete provided by the invention (Streptomyces linzensis) bacterial strain SPC6-50-1.
Fig. 2 is the bacteriostatic activity comparing result schematic diagram of wild-type streptomyces coelicolor (Streptomyces coelicolor A3 (2)) and mutant strain thereof.
Fig. 3 is the bacteriostatic activity comparing result schematic diagram of streptomycete (Streptomyces sp.) Lzsg6 and mutant strain thereof.
Embodiment
Embodiment 1:
By chemical mutagen NTG, streptomycete is carried out to mutagenesis, as shown in Figure 1:
One, the treatment process of NTG mutagenesis bacterial classification:
1, the preparation of substratum and solution used:
TSBY (1L): Oxoid tryptone beans powders (TSB) 30g; Sucrose 340g; Oxoid yeast extract 5g; Distilled water is to 1000ml.
YEME (1L): Oxoid yeast extract 3g; Oxoid peptone Tryptone5g; Malt extract 3g; Glucose 10g; Sucrose 340g; MgCl
2-6H
2o (sterilizing separately); Distilled water is to 1000ml.
PH6.0 phosphoric acid buffer: K
2hPO
42g/L, KH
2pO
48g/L, 0.1MPa sterilizing 20min.
The preparation of NTG mother liquor: take 0.1g NTG and add solubility promoter methane amide or acetone 1mL, then add 0.1mol/L pH6.0 phosphoric acid buffer 9mL, be configured to the NTG mother liquor of 10mg/mL.
2, bacterial classification preparation:
Experimental strain is that streptomyces coelicolor Streptomyces coelicolor A3 (2) separates with Streptomyces sp.Lzsg6(from the husky raw reed Rhizosphere Soil of ecosystem in Linze, Gansu, China (N39 ° 22 ' 02.52 " E100 ° 07 ' 46.68 ")).
Bacterial strain is connected in the test tube that 5mL liquid nutrient medium YEME or TSBY are housed, at 30 DEG C, on the shaking table of 200r/min, cultivates 3-7d, can observe directly obvious cotton-shaped mycelium.
3, the preparation of mutagenesis bacteria suspension:
Get nutrient solution to 2mL doff pipe, the centrifugal 1min of 8000r/min, by the 0.1mol/L phosphoric acid buffer washed twice of pH6.0, is then resuspended in the buffering suspension of 2mL pH6.0.
Two, NTG mutagenic treatment screening:
In bacteria suspension, add NTG mother liquor, making its final concentration is 1mg/mL, processes 60min.
Be disposed, by centrifugal bacteria suspension 8000r/min 1min, by the 0.1mol/L phosphoric acid buffer washed twice of pH6.0, be resuspended in afterwards the buffering suspension of 2mL pH6.0.
Embodiment 2:
Screen by Rifampin the bacterial strain that re-reading system is undergone mutation:
The bacteria suspension that NTG was processed is evenly applied on the Gause I culture medium flat plate that contains 5 μ g/ml Rifampins, cultivate 7-15d for 30 DEG C, the single bacterium colony growing is connected to respectively in the test tube that 5mL liquid nutrient medium YEME or TSBY are housed, and then be prepared into mutagenesis bacteria suspension, carry out NTG mutagenesis, the bacteria suspension of processing is evenly applied on the Gause I culture medium flat plate that contains 10 μ g/ml Rifampins, so repeatedly, carry out 15 μ g/ml, 20 μ g/ml, 25 μ g/ml, 30 μ g/ml, 35 μ g/ml, 40 μ g/ml, 45 μ g/ml, the Rifampin screening of 50 μ g/ml, obtain the insensitive bacterial strain of Rifampin.
Embodiment 3:
The bacterial strain of undergoing mutation by the screening translation system of Streptomycin sulphate:
Garbled Rifampin bacterial strain is carried out to NTG mutagenesis again, method is the same, the bacteria suspension of processing is evenly applied to and contains 5 μ g/mL Streptomycin sulphates, on the Gause I culture medium flat plate of 50 μ g/mL Rifampins, cultivate 7-15d for 30 DEG C, the single bacterium colony growing is connected to respectively in the test tube that 5mL liquid nutrient medium YEME or TSBY are housed, and then be prepared into mutagenesis bacteria suspension, carry out NTG mutagenesis, the bacteria suspension of processing is evenly applied to and contains 10 μ g/mL Streptomycin sulphates, on the Gause I culture medium flat plate of 50 μ g/mL Rifampins, so repeatedly, Concentration of Rifampicin is 50 μ g/mL, Streptomycin sulphate is 15 μ g/ml, 20 μ g/ml, 30 μ g/ml, 40 μ g/ml, 50 μ g/ml, 60 μ g/ml, 70 μ g/ml, 80 μ g/ml, 90 μ g/ml, 100 μ g/ml increase progressively successively, obtain Streptomycin sulphate, Rifampin is insensitive bacterial strain all.
Embodiment 4:
Bacterial strain produces antibiotic detection:
One, the preparation of fermented liquid:
By obtain to Streptomycin sulphate, Rifampin all insensitive bacterial strain at the flat lining out of the Gause I that there is no resistance, collect Conidia preservation, be connected in the triangular flask that liquid nutrient medium YEME or TSBY are housed, at 30 DEG C, on the shaking table of 200r/min, cultivate 3-7d, the centrifugal 1min of 12000r/min, gets supernatant.
Two, Antibacterial Activity:
Supernatant liquor Antibacterial Activity adopts Oxford agar diffusion method.Experimental strain is intestinal bacteria (Escherichia coli) and subtilis (Bacill μ s s μ btilis).The beef extract-peptone agar plate that preparation contains these two kinds of bacterium adds fermented liquid supernatant obtained in the previous step in the cup of Oxford, cultivates 16h for 37 DEG C, by ruler measurement inhibition zone size.Taking the fermented liquid supernatant of wild-type Streptomyces coelicolor A3 (2) and Streptomyces sp.Lzsg6 as contrast, result as shown in Figures 2 and 3.In Fig. 2, taking bacterial strain 2 fermented liquid supernatant to colibacillary antibacterial circle diameter as 100%, bacterial strain 1-9 is 9 mutant strains that Streptomyces coelicolor A3 (2) obtains by this method.In Fig. 3, taking the fermented liquid supernatant of wild-type Streptomyces coelicolor A3 (2) to colibacillary antibacterial circle diameter as 100%, bacterial strain 1-6 is 6 mutant strains that Streptomyces sp.Lzsg6 obtains by this method.
Can find out from the result of Fig. 2 and Fig. 3, after this method is processed, there is very large change in wild-type Streptomyces coelicolor A3 (2) bacteriostatic activity, and the bacteriostatic activity of its mutant strain 2 is about 6 times of wild-type.Itself does not almost have bacteriostatic activity wild-type Streptomyces sp.Lzsg6, and after this method is processed, its mutant strain 2,4,5 has had obvious bacteriostatic activity, illustrates that mutant strain has produced the material with bacteriostatic activity.This explanation this method can activate some silencers of streptomycete, obtains new meta-bolites.
Finally it should be noted that: the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although the present invention is had been described in detail with reference to previous embodiment, for a person skilled in the art, its technical scheme that still can record aforementioned each embodiment is modified, or part technical characterictic is wherein equal to replacement.Within the spirit and principles in the present invention all, any amendment of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (7)
1. streptomycete
(Streptomyces linzensis)the mutagenesis screening method of bacterial strain SPC6-50-1, is characterized in that, is to streptomyces coelicolor by nitrosoguanidine (NTG)
(Streptomyces coelicolor A3 (2))and streptomycete
(Streptomyces sp.)lzsg 6 carries out mutagenesis, obtains streptomycete mutant library, then by Rifampin and Streptomycin sulphate screening, obtains the bacterial strain that re-reading system and translation system are undergone mutation.
2. streptomycete according to claim 1
(Streptomyces linzensis)the mutagenesis screening method of bacterial strain SPC6-50-1, is characterized in that, includes following steps:
1) chemical mutagen NTG carries out mutagenesis to streptomycete: by streptomyces coelicolor (
streptomyces coelicolor A3 (2)) and streptomycete (
streptomyces sp.) Lzsg 6 is prepared as bacteria suspension, with NTG mother liquor, bacteria suspension carried out to mutagenesis, obtains the bacteria suspension that NTG processed;
2) bacterial strain that selects re-reading system to undergo mutation in order to good fortune plansifter: the bacteria suspension that the NTG that step 1) is obtained processed is evenly applied on the Gause I culture medium flat plate that contains 5 μ g/ml Rifampins, cultivate 7-15 d for 30 DEG C, the single bacterium colony growing is connected to respectively in the test tube that 5 mL liquid nutrient medium YEME or TSBY are housed, and then be prepared into mutagenesis bacteria suspension, carry out NTG mutagenesis, the bacteria suspension of processing is evenly applied on the Gause I culture medium flat plate that contains 10 μ g/ml Rifampins, repeatedly, carry out 15 μ g/ml, 20 μ g/ml, 25 μ g/ml, 30 μ g/ml, 35 μ g/ml, 40 μ g/ml, 45 μ g/ml, the Rifampin screening of 50 μ g/ml, obtain the rear bacterial strain of Rifampin screening,
3) bacterial strain of undergoing mutation with Streptomycin sulphate screening translation system: by step 2) bacterial strain carries out NTG mutagenesis again after the Rifampin screening that obtains, method and step 2) identical, the bacteria suspension of processing is evenly applied to and contains 5 μ g/mL Streptomycin sulphates, on the Gause I culture medium flat plate of 50 μ g/mL Rifampins, cultivate 7-15 d for 30 DEG C, the single bacterium colony growing is connected to respectively in the test tube that 5 mL liquid nutrient medium YEME or TSBY are housed, and then be prepared into mutagenesis bacteria suspension, carry out NTG mutagenesis, the bacteria suspension of processing is evenly applied to and contains 10 μ g/mL Streptomycin sulphates, on the Gause I culture medium flat plate of 50 μ g/mL Rifampins, repeatedly, Concentration of Rifampicin is 50 μ g/mL, Streptomycin sulphate is 15 μ g/ml, 20 μ g/ml, 30 μ g/ml, 40 μ g/ml, 50 μ g/ml, 60 μ g/ml, 70 μ g/ml, 80 μ g/ml, 90 μ g/ml, 100 μ g/ml increase progressively successively, obtain Streptomycin sulphate, bacterial strain after Rifampin screening.
3. the streptomycete that a strain obtains with the mutagenesis screening method mutagenesis screening described in claim 1 or 2
(Streptomyces linzensis)bacterial strain SPC6-50-1, is characterized in that, the preserving number of described streptomycete is CGMCC No.8372.
4. streptomycete claimed in claim 3
(Streptomyces linzensis)bacterial strain SPC6-50-1 is in the application of preparing in microbiotic.
5. the streptomycete described in claim 1 or 2
(Streptomyces linzensis)the mutagenesis screening method of bacterial strain SPC6-50-1 is in the application activating in streptomycete silencer.
6. the streptomycete described in claim 1 or 2
(Streptomyces linzensis)the mutagenesis screening method of bacterial strain SPC6-50-1 is in the application of preparing in microbiotic.
7. application as claimed in claim 6, it is characterized in that, in the flat lining out of Gause I by the bacterial strain after Streptomycin sulphate, Rifampin screening, collect Conidia preservation, be connected in the triangular flask that liquid nutrient medium YEME or TSBY are housed, at 30 DEG C, on the shaking table of 200 r/min, cultivate 3-7d, centrifugal 1 min of 12000 r/min, gets supernatant.
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| CN106497981A (en) * | 2016-10-31 | 2017-03-15 | 广东省微生物研究所 | A kind of method of activating microorganisms recessiveness secondary metabolite biological synthesis gene cluster expression |
| CN107475241A (en) * | 2017-09-30 | 2017-12-15 | 上海上药新亚药业有限公司 | A kind of chemical mutation method of streptomysin strain |
| CN109613162A (en) * | 2018-12-05 | 2019-04-12 | 海南省海洋与渔业科学院（海南省海洋开发规划设计研究院） | A method of induction marine actinomycete cryptiogene expression generates novel substance |
| CN114686540A (en) * | 2022-01-21 | 2022-07-01 | 中国科学院西北生态环境资源研究院 | Efficient fermentation preparation process of streptomyces thermalii secondary metabolite |
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| CN107475241A (en) * | 2017-09-30 | 2017-12-15 | 上海上药新亚药业有限公司 | A kind of chemical mutation method of streptomysin strain |
| CN109613162A (en) * | 2018-12-05 | 2019-04-12 | 海南省海洋与渔业科学院（海南省海洋开发规划设计研究院） | A method of induction marine actinomycete cryptiogene expression generates novel substance |
| CN114686540A (en) * | 2022-01-21 | 2022-07-01 | 中国科学院西北生态环境资源研究院 | Efficient fermentation preparation process of streptomyces thermalii secondary metabolite |
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