CN109182367A - MSMEG_6171 is adjusting the application in mycobacterium smegmatis Antibiotic Sensitivity - Google Patents
MSMEG_6171 is adjusting the application in mycobacterium smegmatis Antibiotic Sensitivity Download PDFInfo
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- CN109182367A CN109182367A CN201810907735.0A CN201810907735A CN109182367A CN 109182367 A CN109182367 A CN 109182367A CN 201810907735 A CN201810907735 A CN 201810907735A CN 109182367 A CN109182367 A CN 109182367A
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Abstract
The invention discloses a kind of methods for adjusting mycobacterium smegmatis Antibiotic Sensitivity, it is characterised in that: is realized by adjusting MSMEG_6171 expression quantity.Inhibit MSMEG_6171 expression enhancing mycobacterium smegmatis Antibiotic Sensitivity, promotes MSMEG_6171 to be overexpressed and weaken mycobacterium smegmatis Antibiotic Sensitivity.New method is provided to adjust mycobacterium smegmatis Antibiotic Sensitivity.
Description
Technical field
Field of biotechnology of the present invention, in particular to MSMEG_6171 are adjusting mycobacterium smegmatis Antibiotic Sensitivity
In application.
Background technique
Mycobacterium smegmatis (M.smegmatis) belongs to Grain-positive saprophytic bacteria, have rapid growth, no pathogenicity, with
Mycobacterium tuberculosis (M.tuberculosis) gene very high homology, the features such as eucaryotic cell structure is similar, more application is in branch bar
Bacterium infection and related immunological research, are a kind of experimental models of relative ideal.Meanwhile non-branch bacillus infection and other
Also there is the application of expansion in related immune research.
Mycobacterium smegmatis has the cell wall rich in lipid, and constituent is complicated, and function is also various, is mycobacteria
The pressure of external environment is resisted, the important barrier of host immune attack of escaping.Mycobacterium smegmatis cell wall consists of two parts: interior
Layer and outer layer.The soluble constituent for the cell wall that outer layer is made of lipid and protein, long-chain and short chain fatty acids are inlayed wherein.
Internal layer is made of peptide glycan (PG), arabogalactan (AG), mycolic acid (MA).The study found that more and more albumen
Participate in the synthesis of cell wall, these albumen by participate in the synthesis of fatty acid acyl chain, glycopeptidolipid (GPLs), peptide modification with
And assembling of various synzyme etc. is on cell membrane to influence the generation of cell wall, to influence bacterium to the resistance of drug.Perhaps
Mostly anti-mycobacteria drug is dedicated for this good protective barrier of confrontation.
MSMEG_6171 is the albumen of a Unknown Function in mycobacterium smegmatis, the amino acid sequence of MSMEG_6171
As shown in SEQ ID No.1, encoding gene is as shown in SEQ ID No.2, and there has been no reports for the research of MSMEG_6171 protein function
Road, the homologous protein in mycobacterium tuberculosis are Rv3660c.One of England.K in 2011 is research shows that Rv3660c
It can inhibit diaphragm when overexpression to generate and cause thallus fibrillation, which is analyzed by transcription group, and discovery is overexpressed the egg
Significant changes occur for the white mrna expression amount that can cause some cell regulatory factors, these regulatory factors include suspend mode regulator
It is considered the sigma factors that play a role in adaptability metabolism with some, while it is above-mentioned to find that Rv3660c missing can be offset
The expression quantity of regulatory factor changes, and thallus is caused to shorten.At present there is no MSMEG_6171 and its homologous protein Rv3660c to thin
The relevant report of the adjustment effect of the antibiotics sensitivity of bacterium.
Summary of the invention
The purpose of the present invention is to provide MSMEG_6171 to adjust answering in mycobacterium smegmatis Antibiotic Sensitivity
With.
The technical solution used in the present invention is:
A method of mycobacterium smegmatis Antibiotic Sensitivity is adjusted, it is real by adjusting MSMEG_6171 expression quantity
It is existing.
Further, inhibit MSMEG_6171 expression enhancing mycobacterium smegmatis Antibiotic Sensitivity, promote MSMEG_
6171 are overexpressed decrease mycobacterium smegmatis Antibiotic Sensitivity.
Further, MSMEG_6171 expression quantity is MSMEG_6171 expressing quantity.
Further, antibiotic is the antibiotic for acting on mycobacterium smegmatis Cell wall synthesis.
Further, antibiotic be vancomycin, cefotaxime, Cefoxitin and cephalo pyridine in any one or it is several
Kind.
The reagent for adjusting MSMEG_6171 expression quantity adjusts the reagent of mycobacterium smegmatis Antibiotic Sensitivity in preparation
Application in box.
Further, using the reagent preparation enhancing mycobacterium smegmatis for inhibiting MSMEG_6171 to express to antibiotic sensitive
Property kit, use the reagent preparation for promoting MSMEG_6171 to be overexpressed to weaken mycobacterium smegmatis Antibiotic Sensitivity
Kit.
Further, MSMEG_6171 expression quantity is MSMEG_6171 expressing quantity.
Further, antibiotic is the antibiotic for acting on mycobacterium smegmatis Cell wall synthesis.
Further, antibiotic be vancomycin, cefotaxime, Cefoxitin and cephalo pyridine in any one or it is several
Kind.
The beneficial effects of the present invention are:
The relationship between MSMEG_6171 and mycobacterium smegmatis Antibiotic Sensitivity is disclosed in the present invention, specifically
Ground inhibits MSMEG_6171 expression enhancing mycobacterium smegmatis Antibiotic Sensitivity, promotes MSMEG_6171 to be overexpressed and weaken
Mycobacterium smegmatis Antibiotic Sensitivity.The present invention provides new side to adjust mycobacterium smegmatis Antibiotic Sensitivity
Method.
Detailed description of the invention
Fig. 1 is the PCR primer agarose electrophoresis figure of MSMEG_6171 gene；
Fig. 2 is the mycobacterium smegmatis bacterium solution PCR verifying that MSMEG_6171 is overexpressed, and 1,2,3 swimming lanes are that verifying is positive
Bacterial strain；
Fig. 3 is the Western Blot qualification result of MSMEG_6171-3X Flag expressing fusion protein, swimming lane 1: albumen
marker；Swimming lane 2: zero load control；Swimming lane 3: it is overexpressed recombinant bacterial strain；
Fig. 4 is the mycobacterium smegmatis PCR primer agarose electrophoresis figure of MSMEG_6171 missing, swimming lane 1: deletion mycopremna；
Swimming lane 2: zero load control；M:D2000DNAmarker；
Fig. 5 is Pull Down experimental result；
Fig. 6 is fluorescent staining figure, and wherein A is wild type mycobacterium smegmatis, B is that the shame dirt that MSMEG_6171 is overexpressed divides
Branch bacillus, C are the mycobacterium smegmatis of MSMEG_6171 missing, D is covering bacterial strain.
Specific embodiment
Below in conjunction with specific embodiment, the present invention is further explained.
Embodiment 1
Utilize mycobacteria-E. coli shuttle plasmid pMV261-3X flag (New use of BCG for
recombinantvaccines,Nature,1991.351(6326):456-60；The public can be from Guangdong Province Tubercufosis control center
Obtain) expression MSMEG_6171 albumen, obtain the mycobacterium smegmatis for being overexpressed MSMEG_6171.
One, MSMEG_6171 expression plasmid is constructed
With mycobacterium smegmatis Mycobacterium tuberculosis mc2155 genomic DNA is template, is passed through
Forward primer 5 '-agaattcatgccgtcgagtcgtaagg-3 ' (SEQ ID No.3) and reverse primer 5 '-
Taaaagcttccgccgcctcccgcacag-3 ' (SEQ ID No.4) carries out PCR amplification, obtains MSMEG_6171 gene coding
Segment, detected through gel electrophoresis result is as shown in Figure 1, by EcoRI, HindIII and T4 ligase of NEB company by MSMEG_
6171 genes coding segment is connected to pMV261-3X Flag carrier, obtains MSMEG_6171-3X Flag fusion protein expression plasmid
pMV261-HtpG-3X Flag.Recombinant plasmid is sequenced, confirmation is correct.
Two, pMV261-MSMEG_6171-3X Flag mycobacterium smegmatis is prepared
(1) mycobacterium smegmatis Mycobacterium smegmatis mc2155 bacterial strains containing 0.5% glycerol and
37 DEG C shaken cultivation 3 days in the 7H9 fluid nutrient medium of 0.05% Tween 80,1% switching fresh culture, 37 DEG C of shaken cultivations are extremely
Logarithmic phase (OD600About 0.6), thallus is collected, 10% sterile glycerol (deionized water configuration) cleans 3 preparations of thallus electricity and turns sense
By state.
(2) take 300 μ L of competence be separately added into 10 μ L plasmids (100ng/ μ L) pMV261-HtpG-3X Flag and
PMV261-3X Flag is transferred to electric revolving cup, and setting electric converter parameter is 2.5kV, 25 μ F, 1000 Ω, adds after the completion of electric shock
Enter the fresh culture of 800 μ L pre-cooling, 37 DEG C of shaken cultivation 2h make bacteria resuscitation.
(3) take 1/3 coating containing the 7H10 solid medium of 30 μ g/mL of kanamycin the bacterium of recovery, 37 DEG C quiet
Culture 3 days is set, mycobacterium smegmatis recombinant bacterium pMV261-MSMEG_6171-3X Flag/mc is obtained2155 and pMV261-
3XFlag/mc2155。
(4) picking recombinant bacterium pMV261-MSMEG_6171-3X Flag/mc2155 and pMV261-3X Flag/mc2155
Single colonie, 7H9 fluid nutrient medium of the inoculation containing 30 μ g/mL of kanamycin, 0.5% glycerol and 0.05% Tween 80,37 DEG C
Shaken cultivation 3 days, draw 50 μ L bacterium solution centrifugation thallus；Thallus is resuspended in 10 μ L deionized waters, takes 1 μ L to carry out PCR reaction, draws
Object is the forward primer and reverse primer for constructing MSMEG_6171 expression plasmid and using, and whether verifying MSMEG_6171 gene succeeds
Mycobacterium smegmatis is imported, as a result as shown in Figure 2.
Three, whether expressed using Western Blot detection MSMEG_6171-3X Flag fusion protein
Take 20 μ g pMV261-MSMEG_6171-3X Flag/mc2155 cell pyrolysis liquids carry out Western Blot inspection
It surveys, as a result as shown in figure 3, the fluorescence signal that may be significantly, it was demonstrated that MSMEG_6171-3X Flag is expressed really.
Embodiment 2
MSMEG_6171 gene knock-out bacterial strain is constructed by the method that homologous gene recombinates.
1. homologous gene of table recombinates relevant recombinant plasmid
One, the building of suicide plasmid
(1) PCR amplification MSMEG_6171 upstream and downstream homology arm: by MSMEG_6171 gene and its upstream and downstream DNA sequence dna, if
Count primer 6171-UF:5 '-cccaagcttatgtagtcacgcatccggtcc-3 ' (SEQ ID No.5) (underlined sequences table
Showing indicates the site HindIII), 6171-UR:5 '-gagtgcctcgtcgagacccggttcgtcgcatccgatcac-3 ' (SEQ
ID No.6), using M.smegmatis genomic DNA be template amplification obtain MSMEG_6171 upstream region of gene homology arm (Up,
855bp).With 6171-DF:5 '-gtgatcggatgcgacgaaccgggtctcgacgaggcactc-3 ' (SEQ ID No.7) and
(underlined sequences indicate PacI to 6171-DR:5 '-ttaattaacggtgttgagcgcggcg-3 ' (SEQ ID No.8)
Point), using M.smegmatis genomic DNA be template amplification obtain MSMEG_6171 downstream of gene homology arm (Down,
849bp).Upstream and downstream homology arm segment distinguishes purification and recovery, using the method for overlap PCR, with upstream homology arm and downstream
Homology arm is template, using 6171-UF and 6171-DR as primer amplification overlapping fragments.
(2) HindIII and PacI restriction enzyme site is utilized, the upstream and downstream homology arm connected is inserted into p1NIL carrier, then
It will be recycled after the genetic fragment PacI digestion for being used to screen in pGOAL19 carrier, then this segment be connected into containing homology arm
P1NIL recombinant vector in.
(3) suicide plasmid is transferred to E.coli Top10, PCR and sequence verification, upgrading grain is spare.
Two, alkali process suicide plasmid
It takes a small amount of preservation plasmid to measure its concentration, and takes the plasmid of 20 μ g or so according to its concentration, add ddH2O to 100 μ
Then the NaOH of 100 μ L 0.4mol/L is added in L.37 DEG C, 30min.140 μ L isopropyls are added after the 3M sodium acetate of 20 μ L is added
Alcohol, -80 DEG C of precipitating 1h or overnight, supernatant is abandoned in centrifugation, adds 700 μ L, 75% ethyl alcohol, and supernatant is abandoned in centrifugation, and repeated washing is primary.Room temperature
After standing 5min, add 7 μ L ddH2O dissolution.
Three, single exchange strains are screened
Plasmid after 7 μ L alkali process is added in the competent cell of 350 μ L to (note: 200 μ L competent cells institutes are right
Plasmid volume is answered to be no more than 5 μ L), then bacterium solution is transferred in electric revolving cup by 37 DEG C of 10~20min of incubation.Confirm electric revolving cup outer wall
It after going up completely, shocks by electricity (voltage 2.5kV, 1000 Ω of resistance, 25 μ F of capacitor), the 7H9 fluid nutrient medium of 0.7mL is added immediately.It will
Thallus is sucked out, and is placed in 50mL centrifuge tube, and adding 7H9 fluid nutrient medium, (usual recovery used medium is competence to 4mL
10 times of volume).It recovers within shaken cultivation 2~3 days at 37 DEG C.After recovery, 6000rpm is centrifuged 15min, and 200 μ L is taken to cultivate base weight
Outstanding thallus, coating 7H10 plate (contain OADC, 50 μ g/mL hygromycin, 50 μ g/mL kanamycins and 50 μ g/mLXgal), are protected from light
Culture has seen whether locus coeruleus generation in 30~40 days, if so, then showing single-swap success.
Four, double crossing over bacterial strain is screened
It is protected from light culture 30~40 days and has seen whether locus coeruleus generation, if so, then showing single-swap success, locus coeruleus is chosen, in 7H9
It is cultivated in fluid nutrient medium.200 μ L are taken to apply 7H10 plate (containing OADC, X-Gal, 2% in 4h, 8h, 12h, 16h respectively
sucrose).Be protected from light in 37 DEG C of incubators culture 2-3 days, picking white colony, cultivated in 2mL 7H9 culture medium 2 days it is laggard
Row PCR verifying.
Five, mycobacterium smegmatis MSMEG_6171 deletion mycopremna covers
(1) covering plasmid is pMV361-MSMEG_6171.
(2) preparation of mycobacterium tuberculosis MSMEG_6171 deletion mycopremna competent cell: above-mentioned shame dirt branch bar is referred to
The preparation of bacterium competence cell.The mycobacterium smegmatis PCR qualification result of MSMEG_6171 missing is as shown in Figure 4.
(3) competent cell is electroporated: referring to above-mentioned step of converting.
Embodiment 3
Wild type mycobacterium smegmatis, the mycobacterium smegmatis of MSMEG_6171 missing, covering bacterial strain and MSMEG_6171
The mycobacterium smegmatis of overexpression detects the MIC of 10 kinds of antibiotic.
Using dilution method detection MSMEG_6171 to the sensibility of antibiotic.By the antibiotic of various concentration after doubling dilution
Solution (rifampin, isoniazid, streptomysin, vancomycin, 2-ethylisonicotinthionamide, seromycin, Meropenem, cefadole, cephalo
Western fourth, cefotaxime) it is added separately in 96 sterile hole polystyrene plates.1st to the 11st hole dosing, every 10 μ L of hole, the 12nd hole
Not dosing is used as growth control, 3 parallel samples.After bacterium solution is diluted to 0.5 maxwell reduced turbidity, then with 7H9 fluid nutrient medium
After 1: 1000 dilution, bacterium solution to after every 90 μ L of Kong Zhongjia dilution, 37 DEG C of shaking tables of sealing postposition are incubated for 2~3h, are able to suppress
The minimum concentration of bacterial growth is MICs.
It is different through the measurement result of the minimum inhibitory concentration (MIC) of 10 kinds of different antibiotic it is found that for all bacteriums
Cigarette hydrazine, rifampin, streptomysin, seromycin are in same level.After MSMEG_6171 gene delection, mycobacterium smegmatis pair
The sensibility enhancing of vancomycin, cefotaxime, Cefoxitin and cephalo pyridine, corresponding MIC value reduce；Meanwhile MSMEG_
6171 mycobacterium smegmatis being overexpressed weaken the sensibility of vancomycin, cefotaxime, Cefoxitin and cephalo pyridine, right
The MIC value answered increases.
Embodiment 4
The albumen that Pull Down experiment is fished in mycobacterium smegmatis with MSMEG_6171 interaction, as a result as shown in figure 5,
By the protein in Mass Spectrometric Identification compound with MSMEG_6171 interaction after differential band is digested, by MSMEG_
The analysis of 6171 interaction proteins finds that the cell walls such as MSMEG_6171 and fatty acid synthetase, glycerol-3-phosphate dehydrogenase close
Exist at relevant enzyme and interact, illustrates that MSMEG_6171 as a member in albumen composition, participates in bacteria cell wall
Synthesis.
Embodiment 5
Whether fluorescent staining verification experimental verification MSMEG_6171 will affect the formation of the nucleus and diaphragm of mycobacterium smegmatis.
After the vancomycin of BODIPY label and DAPI dyeing, carried out with Zeiss LSM880 confocal microscope
Observation, as a result as shown in Figure 6.By to wild type mycobacterium smegmatis, MSMEG_6171 overexpression mycobacterium smegmatis,
MSMEG_6171 missing mycobacterium smegmatis and cover bacterial strain fluorescent staining discovery, four kinds of bacterium cultivate in vitro lower diaphragm plate and
The no notable difference of the generation of nucleus illustrates that MSMEG_6171 does not have the generation of the diaphragm and nucleus of mycobacterium smegmatis
It influences.
SEQUENCE LISTING
<110>Guangdong Province's Tubercufosis control center
<120>MSMEG_6171 is adjusting the application in mycobacterium smegmatis Antibiotic Sensitivity
<130> 2018.8.10
<160> 8
<170> PatentIn version 3.5
<210> 1
<211> 316
<212> PRT
<213> MSMEG_6171
<400> 1
Met Pro Ser Ser Arg Lys Ala Trp Leu Ser Ala Ala Ala Val Leu Leu
1 5 10 15
Asp Ala Ser Ala Ala Arg Arg Cys Ala Asp Arg Gly Leu Pro Arg Arg
20 25 30
Asp Arg Val Leu Val Ile Gly Cys Asp Glu Pro Gly Pro Ala Asp Trp
35 40 45
Gln Ala Ala Ile Ala Val Gly Ala Gln His Val Val Thr Leu Pro Arg
50 55 60
Gln Asp Thr Asp Leu Val Ala Ala Leu Ser Val Val Asp Glu Gly Gly
65 70 75 80
Gly His Arg Gly Pro Val Val Ala Val Val Ala Ala Lys Gly Gly Ala
85 90 95
Gly Ala Ser Val Phe Ala Ala Ala Leu Ala Leu Ser Ala Pro Gly Ala
100 105 110
Leu Leu Val Asp Ala Asp Pro Trp Ser Gly Gly Ile Asp Leu Val Leu
115 120 125
Gly Ser Glu Asp Gln Pro Gly Leu Arg Trp Ala Asp Leu Ala Leu Gln
130 135 140
Gly Gly Arg Leu Gly Tyr Gly Ala Leu Arg Asp Ala Leu Pro Arg Arg
145 150 155 160
Gly Glu Ile Ser Val Leu Ser Gly Gly Arg Ala Gly Val Asp Ile Thr
165 170 175
Ala Ala Ala Leu His Ala Val Ile Asp Ala Gly Cys Arg Gly Ala Thr
180 185 190
Leu Val Val Cys Asp Val Pro Arg Arg Ser Thr Asp Ala Ala Glu Ala
195 200 205
Ala Leu Glu Ala Ala Asp Leu Val Val Val Val Ala Arg Ala Asp Val
210 215 220
Arg Ser Cys Ala Ala Ala Ala Ala Ala Gly Ser Trp Ile Ala Thr Cys
225 230 235 240
Asn Pro Asn Ile Gly Val Val Val Arg Gly Pro Ala Pro Gly Gly Leu
245 250 255
Arg Ala Ala Glu Val Ala Asp Ile Val Asp Leu Pro Leu Leu Ala Ser
260 265 270
Met Arg Pro Gln Pro Gly Leu Asp Glu Ala Leu Glu Arg Gly Gly Leu
275 280 285
Arg Leu Thr Arg Arg Ser Pro Leu Ala Thr Ala Ala Arg Arg Val Leu
290 295 300
Gly Val Leu Ala Gln His Pro Val Arg Glu Ala Ala
305 310 315
<210> 2
<211> 812
<212> DNA
<213> MSMEG_6171
<400> 2
atgccgtcga gtcgtaaggc ctggctgtcc gcggcggccg tgctgctcga cgcgtcggcc 60
gcgcggcggt gcgccgaccg gggactgccg cgccgcgacc gggtgctcgt gatcggatgc 120
gacgaaccgg gtccggccga ttggcaggcc gcgatcgccg tcggtgcgca acacgtcgtg 180
acgctgcccc gccaggacac cgatcttgtg gcggcgctgt ctgtcgtcga cgagggcggt 240
ggacaccgcg gaccggtggt tgcggtggtc gcggcgaaag gcggcgccgg ggcatcggtg 300
ttcgccgccg cgcttgccct gtcggcgccg ggagcgttgc tcgtcgacgc cgacccgtgg 360
agcggcggca tcgacctcgt gctcggcagt gaggatcagc cgggtctgcg gtgggcggat 420
cttgcgctgc agggcggcag gctcgggtac ggcgcgttgc gcgacgcgct gccgcgccgc 480
ggcgagatca gtgtgctctc gggtggacgc gcgggcgtcg acatcaccgc ggcggcactg 540
cacgccgtga tcgacgcggg ttgtcgcggc gcgactttgg tggtctgcga cgtcccccgg 600
cgcagcaccg acgcggccga ggccgcgctc gaggcagccg acctggtggt cgtggtcgcc 660
cgcgccgatg tgcgttcgtg tgccgcggca gcggccgccg gatcctggat cgccacgtgc 720
aatcccaaca tcggtgtggt ggtgcgcggt ccggcaccgg gaggtctgcg tgccgccgag 780
gtcgccgaca tcgtcgacct gccgctgctg gc 812
<210> 3
<211> 26
<212> DNA
<213>artificial synthesized
<400> 3
agaattcatg ccgtcgagtc gtaagg 26
<210> 4
<211> 27
<212> DNA
<213>artificial synthesized
<400> 4
taaaagcttc cgccgcctcc cgcacag 27
<210> 5
<211> 30
<212> DNA
<213>artificial synthesized
<400> 5
cccaagctta tgtagtcacg catccggtcc 30
<210> 6
<211> 39
<212> DNA
<213>artificial synthesized
<400> 6
gagtgcctcg tcgagacccg gttcgtcgca tccgatcac 39
<210> 7
<211> 39
<212> DNA
<213>artificial synthesized
<400> 7
gtgatcggat gcgacgaacc gggtctcgac gaggcactc 39
<210> 8
<211> 25
<212> DNA
<213>artificial synthesized
<400> 8
ttaattaacg gtgttgagcg cggcg 25
Claims (10)
1. a kind of method for adjusting mycobacterium smegmatis Antibiotic Sensitivity, it is characterised in that: by adjusting MSMEG_6171
Expression quantity is realized.
2. according to the method described in claim 1, it is characterized by: inhibiting MSMEG_6171 expression enhancing mycobacterium smegmatis pair
Antibiotics sensitivity promotes MSMEG_6171 to be overexpressed and weakens mycobacterium smegmatis Antibiotic Sensitivity.
3. according to the method described in claim 1, it is characterized by: MSMEG_6171 expression quantity is MSMEG_6171 protein expression
Amount.
4. according to the method described in claim 1, it is characterized by: antibiotic is to act on mycobacterium smegmatis Cell wall synthesis
Antibiotic.
5. according to the method described in claim 1, it is characterized by: antibiotic be vancomycin, cefotaxime, Cefoxitin and
Any one or a few in cephalo pyridine.
6. the kit that the reagent for adjusting MSMEG_6171 expression quantity adjusts mycobacterium smegmatis Antibiotic Sensitivity in preparation
In application.
7. application according to claim 6, it is characterised in that: enhanced using inhibiting the reagent of MSMEG_6171 expression to prepare
The kit of mycobacterium smegmatis Antibiotic Sensitivity weakens shame dirt using the reagent preparation for promoting MSMEG_6171 to be overexpressed
The kit of mycobacteria Antibiotic Sensitivity.
8. application according to claim 6, it is characterised in that: MSMEG_6171 expression quantity is MSMEG_6171 protein expression
Amount.
9. application according to claim 6, it is characterised in that: antibiotic is to act on mycobacterium smegmatis Cell wall synthesis
Antibiotic.
10. application according to claim 6, it is characterised in that: antibiotic is vancomycin, cefotaxime, Cefoxitin
With any one or a few in cephalo pyridine.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101923091A (en) * | 2010-05-18 | 2010-12-22 | 青岛瑞杰生物科技有限公司 | Method for detecting high-sensitivity mycobacterium tuberculosis |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101923091A (en) * | 2010-05-18 | 2010-12-22 | 青岛瑞杰生物科技有限公司 | Method for detecting high-sensitivity mycobacterium tuberculosis |
Non-Patent Citations (2)
| Title |
|---|
| PERRODOU,E ET AL: "ACCESSION NO.AFP42444，hypothetical protein MSMEI_6012 [Mycolicibacterium smegmatis MC2 155]", 《GENBANK》 * |
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