METHOD FOR PREPARING GRAM-POSITIVE BACTERIAL GHOSTS BY HYDROCHLORIC ACID TREATMENT
The present invention refers to gram negative bacteria prepared by the number by the hydrochloric acid bath method are disclosed to ghost and number. Bacterial ghost (bacteria ghost, bacterial ghost) (cytoplasmic inclusions) number to a microbial cells of compositions is a stand-alone, but the form of the outer membrane (envelope) complete the interior case that the extracted cell membranes can be simply defining structural body can be maintained. Bacterial ghost which lack an intracellular DNA or dielectric material, substantially sterilising thickness as the gene recombinant organisms (GMO) is not considered. However, the bacterial ghost form passageways since maintaining a locus is canceled by the present hepatitis c (antigenic determinant) is judged to be the outer membrane, with lifestyle vaccine can be functionally similar effect. In particular, in the treatment of a bacterial infection, vaccines and immunotherapeutics using bacterial ghost (ghost vaccine) quantities of non-specific immune induction the inside due to the cytoplasmic free, the catabolic pathway (adjuvant) without adding foreign simultaneous auxiliary number can be easily stimulating immune and adaptive immune system. Due to such benefit, while bacterial ghost vaccine using low cost effective limit of the existing method overcomes the chemicals polynucleotide can be alternatively the pupil disclosed. In addition, the locus of the proliferation or pathogenic bacterial ghost as in memory, deactivated (inactivated) state of bacterial ghost animal, human or plant tissue or cell can be attached to be special. As well as, recombination can be introduced into plant cells or animal cells to target cells can be effectively transferred or nucleic acid antigen delivery system (delivery system) can be dissolved. The various method for the production of bacterial ghost developed in the nanometer range. The most general method for producing a bacterial ghost E protein mediated melting method as, a plasmid (plasmid) gene E φ X174 bacteriophage (bacteriophage) dissolved is useful in a method to gram-negative bacteria to transformed (transformation) followed by the expression thereof are disclosed. Transformed bacteria of a billion number E gene response fuel E which protein is expressed, synthesized E protein bacteria without damaging the bacterial cell membranes and cell-wall MLV physicochemical surface structure with excessive cell composition water tunnel release other (non-patent papers 0001). In this method, when a desired protein antigen gene can be various plasmid cloning change, mass spectrometry mass production through existing gene for diverter pin is accumulated. However, said E protein mediated dissolution method including, herpes simplex virus transformed bacterial ghosts through bath method is useful in number for the, multi-stage process involving molecular biological molecules of one of the pin is the disadvantage that cost and long-term number bath time. The, simple number bath technique, the low-cost production costs and time can be saved, etc. bacterial ghosts required mass production method. Recently, gram-negative bacteria is Escherichia coli ( Most bacterial ghost and e. coli or Salmonella vaccines ( However, non-intended to be of crystallizer antigen protein capable of liberating a process according minerals can occur since, when basic sodium hydroxide is processed by the bacterial ghosts as vaccines that is prepared by the number can be reduced is bigger disclosed. In order to overcome this, studies of bacterial ghosts even number tank supplying acidic environment but request, acid (acid) improves patent studies using bacteria is reported bar free. , the bacteria of the present invention the victims of the gram acid bath method to perform ghost number effort results, provided is a gram-positive bacteria Listeria to gap toe four [su ( , the present invention through bacterial ghosts to victims of the processing of the present invention high pressure liquid coolant hydrochloride Listeria to gap toe four [su number the arrears of work. Thus, the purpose of the invention is, a gram-positive bacterial ghosts a number [...] number bath method are disclosed. Of the present invention another object is to provide, a gram-positive bacterial ghosts or hypermetropia [...] number the number produced therewith said method. Of the present invention another object is to provide, a gram-positive bacterial ghost said active ingredient including, for prevention and treatment of infections gram number [...] vaccine compositions are disclosed. In order to achieve said purposes, the present invention refers to I) incubating perpendicularly installed gram-positive bacteria; Ii) said step i) are cultured in harvested from the gram to yield; Iii) said step ii) hydrochloric acid to form a bacterial ghost gram negative and positive bacteria obtained in processing; and Iv) said step iii) obtain including ghosts formed in bacteria, a gram-positive bacterial ghosts number bath method number [...] substrate. In the embodiment of the present invention according to one preferred, said step i) of gram-positive bacteria [...] ( In the embodiment of the present invention according to one preferred, said step iii) treatment with the treatment of the hydrochloric acid concentration (MIC) be a minimum billion number. In the embodiment of the present invention according to one preferred, said minimum billion number concentration hydrochloric acid 6. 25 mg/Ml can be one having an. In the embodiment of the present invention according to one preferred, said step iii) to which the valve is 10 to 60 minutes can be the treatment of the hydrochloric acid. In the embodiment of the present invention according to one preferred, said step iii) can be 30 to 40 °C to which the valve is in the treatment of the hydrochloric acid. The present invention refers to in addition, gram-positive bacterial ghosts number to said number tank method number produced therewith [...] substrate. The present invention refers to in addition, said active ingredient including a gram-positive bacterial ghost, vaccine compositions for prevention and treatment of infections gram number [...] substrate. In the embodiment of the present invention according to one preferred, said [...] gram-positive bacteria ( In the embodiment of the present invention according to one preferred, said carrier including bacterial ghosts as foreign antigen or vaccine compositions [...] scene may be disclosed. Thus, the present invention refers to a corrosion treatment gram bacterial ghost and manufacturing method number [...] substrate. The number of gram-positive bacterial ghost gram of the present invention can be hydrochloric acid concentration (MIC) minimum billion billion locus colony generation number processing when cultured, bacterial ghost can be formed effectively. In addition, formed said outer membrane of living bacteria form while preserving mammalian protein or DNA which controls the ghost distance because it has not, to produce a prolonged stress means such as when proliferating 2 difference infection risk. Thus, the gram-positive bacteria of the present invention for prevention or treatment of gram-positive infections ghost vaccines or foreign antigen carrier is useful as the can. Figure 1 Listeria to gap toe four [su ( Hereinafter, the present invention more detailed as follows. As above-mentioned, in bacterial ghost number number through treatment with a chemical bath method, basic processing includes the first number a method publicly known bacterial ghost gram sodium hydroxide bath is the bottom of the sea, acid processing on gram bacterial ghost manufacturing method achieved studies have no disclosed. The recombinant expression vector of the present invention according to hydrochloric acid through a number of gram-positive bacterial ghost gram produced therewith can be hydrochloric acid concentration (MIC) minimum billion billion colony generation locus number number processing when cultured, effectively bacterial ghost can be formed, said formed outer membrane of living bacteria form while preserving mammalian protein or DNA which controls the ghost distance because it has not, when proliferating 2 such as infections for a prolonged difference to produce a normal risk, of the present invention for prevention or treatment of a gram gram of bacterial ghost foreign antigen carrier to use efficient vaccines or infections. Thus, the present invention refers to I) incubating perpendicularly installed gram-positive bacteria; Ii) said step i) are cultured in harvested from the gram to yield; Iii) said step ii) hydrochloric acid to form a bacterial ghost gram negative and positive bacteria obtained in processing; and Iv) said step iii) obtain including ghosts formed in bacteria, a gram-positive bacterial ghosts number bath method number [...] substrate. Of the present invention step i) of the [...] "gram" ( Specifically, said [...] ( Of the present invention step i) of the "culture", preferably 30 to 40 °C culture in gram-positive bacteria, more preferably cultured in specifically 37 °C. In addition, preferably 65 to 75 hours culture of gram-positive bacteria, specifically 72 beany more preferably. Said gram in culture, bacterial growth step (exponential growth phase) hydrochloric acid sensitive organelles of a wall but the logarithm of dish use, use of hydrochloric acid (stationary phase) with respect to the stagnation of the dish after log for MIC is a high layer of the outer membrane, the cell wall have an elastic, dissolved effective tunnel structure are disclosed. Said step iii) hydrochloric acid concentration (MIC) treatment with the minimum number of "processing" is preferably billion but, is not limited. Specifically, hydrochloric acid concentration is preferably 6 to 7 mg/Ml number in said minimum billion, more specifically 6. Most preferably 25 mg/Ml mm.. The present invention according to a gram-positive bacterial ghosts in number bath method, minimum billion number concentration hydrochloric acid processing is important disclosed. MIC hydrochloric acid at a concentration of less than perfect dissolution rate of survival without treatment of gram can be a bacteria, bacterial ghosts [...] scene or foreign antigen carrier cannot used as the number produced therewith. In addition, treatment of the hydrochloric acid concentration greater than MIC gram of damage and increased levels of the passageways, intended to complete tunnel structure form of bacterial ghosts in a clock number cannot be is managed by high pressure liquid coolant, can reduce foreign antigen [...] scene or as a carrier. Said step iii) is 10 to 60 minutes of "processing" to which the valve is preferably hydrochloric acid, specifically 15 to 60 minutes or more preferably processing, is not limited. A gram-positive bacterial ghosts in number bath method of the present invention, the dissolution rate of 100% after about 15 minutes after an MIC of hydrochloric acid can be gram level. But, bacterial ghosts if considering safety in use as vaccines, most preferably 60 minutes to process said processing of the hydrochloric acid. Said step iii) of hydrochloric acid is preferably 30 to 40 °C the processing of "processing" but, not limited to, specifically 37 °C to which the valve is in most preferably. Said processing temperature can be less than 30 °C 40 °C is prevented, are non-MIC is corrosion, bacterial ghost number bath can be the emission efficiency of the present invention. In the embodiment of the present invention in particular, the present invention bacterial ghost number for the victims of the bacteria Listeria to gap toe four [su MIC results identify high pressure liquid coolant of hydrochloride, hydrochloric acid 6. Added 25 mg/Ml concentration when growth of Listeria bacteria is effectively billion number, determined when the same MIC corrosion, the degree of growth of the number has been confirmed that the locus effectively billion (also 1A and also 2). Controls comparisons of sodium hydroxide used for MIC is 6. 25 mg/Ml or, bacterial ghosts for gram-negative bacteria are capable of ammonium sulfate or calcium chloride concentration of 500 mg/Ml number reported high pressure liquid coolant even number didn't effect the growth billion (also 1B to 1D). In addition, the bacteria of Listeria bacteria ghost number in the bath, MIC processing result confirming the optimum condition of hydrochloric acid, hydrochloric acid processing about 15 minutes of MIC effect has been confirmed that the completed the bacterial ghosts (3 also). In addition, the recombinant expression vector of the present invention parties according of the present invention method can be used effectively determines number prepared by the Listeria bacteria ghost result, said bacterial ghost cells to efficiently produce dissolved wall tunnel and a twisted mammalian genome DNA to proteins and devoid of remaining, bacterial ghosts holding structure in the presence of only cell wall protein has been confirmed (also 4 and 5 also). While, in the case of mammalian sodium hydroxide processing number is the same as the high pressure liquid coolant remaining impurities through genome DNA is bacterial ghost, through processing method may be more effective corrosion has been confirmed (6 also). In addition, the present invention through a number of victims of the hydrochloride or sodium hydroxide prepared by the Listeria bacteria surface analysis results of bacterial ghosts, tunnel structure on the bacterial ghost dissolved in hydrochloric acid processing ports, to remain in the pathogenic effects related to somewhat dissolved form has been confirmed (also 7). The, of the present invention method number according to the number of gram-positive bacterial ghost gram produced therewith can be hydrochloric acid concentration (MIC) minimum billion billion locus colony generation number processing when cultured, effectively bacterial ghost can be formed, said formed outer membrane of living bacteria form while preserving mammalian protein or DNA which controls the ghost distance because it has not, when proliferating 2 such as infections for a prolonged difference to produce a normal risk, according to the number of the present invention method for prevention or treatment of infections of bacterial vaccines or ghost gram gram produced therewith foreign antigen carrier as can be used effectively. In addition, the present invention refers to of the present invention a gram-positive bacterial ghosts bath method number according to the number produced therewith, a gram-positive bacterial ghosts number [...] substrate. In addition, the present invention refers to said active ingredient including a gram-positive bacterial ghost, vaccine compositions for prevention and treatment of infections gram number [...] substrate. The [...] "gram" of the present invention ( Specifically, said [...] ( "Vaccine composition" of the present invention of the present invention is new or foreign antigen carrier preferably bacterial ghosts [...] gram as including but, not limited to. The number of gram-positive bacterial ghost gram of the present invention can be hydrochloric acid concentration (MIC) minimum number billion billion locus colony generation processing can be formed when cultured effectively bacterial ghost, formed said outer membrane of living bacteria form while preserving mammalian protein or DNA which controls the ghost distance because it has not, when proliferating 2 such as infections for a prolonged difference to produce a normal risk, gram-positive bacteria of the present invention for prevention or treatment of gram-positive infections as the ghost foreign antigen vaccine or carrier can be used effectively. Hereinafter, the present invention broadcast receiver through more detailed in the embodiment. In the embodiment for the present invention is only to exemplify these provided, in the embodiment of the present invention interprets these range by one number is not in the person with skill in the art will-case in the art. ListeriaMono to gap toe four [su( <1-1> Bacteria Ghost The minimum number jaws billion number concentration (minimum inhibition concentration, MIC) For identifying In the present invention in bacteria Listeria to gap toe four [su bacterial ghost number used in bath, MIC required corrosion therefrom. Specifically, BHI (brain heart infusion) provided is a gram-positive bacteria in a medium 7 °C, 200 rpm conditions because during night (overnight) culturing, Biochrom Libra S22 to O spectrophotometer (spectrophotometer). D. 600 nm measured in absorbance of microbial growth degree therefrom. Then, hydrochloric acid [table 1] new BHI medium such as dilution and concentration to step, said Listeria culture bacteria to gap toe four [su 106 The seeding CFU/Ml concentration, in 18 hours 37 °C him as. Culture nor post-incubation, hydrochloric acid concentration according to degree of microbial growth Listeria to gap toe four [su, O. D. 600 nm measured in absorbance has been confirmed. The positive controls for use, existing bacterial ghost number bath method reported in ammonium sulfate, calcium chloride and sodium hydroxide [table 1] to have a concentration of BHI contains, hydrochloric BHI and adhesive not regulated nothing control processing, said method identical to the Listeria to gap toe four [su the degree of microbial growth therefrom. As a result, as shown in Figure 1, BHI liquid medium 6 sodium hydroxide or hydrochloric acid. 25 mg/Ml concentration when added, each free medium pH pH 3. 16 and pH 10. 75 through a pulse, the growth of Listeria bacteria effectively billion number, Listeria bacteria of MIC 6 is vapourised hydrochloric acid and sodium hydroxide. About 25 mg/Ml (also 1A and 1B) has been confirmed. The compared, the number of e. coli bacterial ghosts to gram-negative bacteria to existing growth billion number number reported to ammonium sulfate and calcium chloride are effective for chemical bath, even concentration of 500 mg/Ml number effect against gram-positive bacteria Listeria billion shows growth has been confirmed (also 1C and 1D). <1-2> Listeria bacteria For bacteria Ghosts Locus Growth ability Confirmation For gram-positive bacteria Listeria bacteria since the MIC of hydrochloric acid and sodium hydroxide obtained by identifying, MIC of hydrochloric acid or sodium hydroxide is processed in a medium number bacterial ghosts standard analysis method (plating) locus has been confirmed that the legal entity to herpesvirus growth degree produced therewith. Specifically, said in the embodiment <1-1>In Listeria bacteria cultured in, such as herpesvirus BHI agar medium concentration for [table 2] then, derived by culturing colony formation of 37 °C 18 in time then, observed. As a result, as shown in Figure 2, nothing control controls a dilution 1/1000 dilution in culture medium to which the corrosion to disenable measurement drain braided with one another to such an extent that many colony herpesvirus, 3. 125 mg/Ml 1/10 dilution concentration of hydrochloric acid processing group to work range of colony formed medium herpesvirus drain dilution but making sure that the crushing, MIC in 6. 25 mg/Ml concentration of hydrochloric acid processing group without dilution medium has been confirmed that the little colony survival in human herpesvirus (also 2A) respectively. In addition, nothing control controls a drain dilution 1/1000 dilution even sodium hydroxide processing group characterized in that it has numerous colony is generated by human herpesvirus in a medium, 3. The concentration of sodium hydroxide solution processing group 125 mg/Ml medium at least one colony formed but making sure that the human herpesvirus work range transition, MIC in 6. 25 mg/Ml concentration of sodium hydroxide solution processing group to mark has been confirmed that the little colony survival in culture medium to which the human herpesvirus (also 2B). <1-3> Listeria bacteria For bacteria Ghost For numerical control machine number, determination of optimum time In hydrochloric acid and sodium hydroxide of MIC against gram-positive bacteria Listeria bacteria obtained by high pressure liquid coolant ghosts can be number checks, bacterial ghost number has been confirmed that the minimum time for numerical control machine. Specifically, 72 hours after culturing centrifuging Listeria bacteria inoculated BHI liquid medium (10, 000g, 10 minutes) and obtained by, phosphate buffer solution (PBS, pH 7. 0) after washing, 106 CFU/Ml Listeria bacteria concentration average value regulated so as to work through. Then, 12. 2 ml Listeria bacteria mixed with hydrochloric acid or sodium hydroxide concentration of 5 mg/Ml preparing final hydrochloride concentration 6. In culturing may be 37 °C was 25 mg/Ml to Listeria. After 15, 30, 45, 60 and 90 component Listeria bacteria culture disclosure in high purity, measured dissolution rate, said in the embodiment <1-2>The method identical to the method of microbial colony forming unit (Colony Forming Unit, CFU) human herpesvirus performed Listeria therefrom. After the culture, Listeria bacteria in high purity PBS solution then-wash 2, 10, 000g bacterial ghosts in 15 minutes centrifuging the resulting Listeria to gap toe four [su obtained. Disclosure 5, 10, 15, 30, 45 and 60 minutes after sodium hydroxide have your culture bacteria in high purity, Listeria bacteria bacterial ghosts number has been confirmed that the numerical control machine. As a result, as shown in Figure 3, MIC of 15 minutes in hydrochloric acid hydrochloric acid processing group processing bacterial ghost is effectively formed, processing 15 minutes BHI agar medium colony layer pattern since the bacteria prepared by the number ghosts herpesvirus (also 3A) has been confirmed. In addition, the MIC of bacterial ghosts about 10 minutes sodium hydroxide sodium hydroxide processing group completed the high-K dielectric oxide or number, referred to as a human herpesvirus BHI agar medium [...] prepared by the number 10 minutes processing since ghosts respectively colony has been confirmed (also 3B). ListeriaMono to gap toe four [su Bacteria Ghosts Identifying features <2-1> Bacteria Ghost Identification of protein amount remaining in Sodium hydroxide or hydrochloric acid number for identifying bacterial ghosts to Listeria prepared by the recombinant expression vector of features, has been confirmed that the amount of protein remaining in pak reel or ghost. Specifically, said in the embodiment <1-3>The method of 15, 30, 45 and 60 minutes after adding hydrochloric acid or sodium hydroxide Listeria bacteria in high purity, (Laemmli, 1970, Nature 227:680 - 685) modified buffer adding, heated 3 to 5 minutes, modified sample Royal. 4 loading time in preparing gel sample is 12% SDS-a PAGE 40 mA current conducting SDS-a PAGE deploy electrophoretic analysis. The brilliant blue R-a 250 including a complete deployment and use in coma of gel (methanol: acetic acid: water=5:1: 5 (v: v: v)) to solution, bacterial ghost in total protein amount has been confirmed. Nothing control controls include, e. coli method identical to the total protein amount in said performing electrophoresis SDS-a PAGE objects has been confirmed that the As a result, as shown in Figure 4, whilst remaining verify mainly hydrochloric acid processing device improves polymer protein in bacteria, mainly low molecular weight protein in sodium hydroxide processing device when the remaining verify, nothing control is shown in e. coli protein band as compared to small troop contrast by neutrophils (4 also). Through, and the mammalian protein in the present invention number prepared by the bacteria present in ghost lack, bacterial ghosts holding structure has been confirmed that the only remaining cell-wall protein present. <2-2> Bacteria Ghost Remaining inside the DNA Amount of Confirmation In the present invention number since protein loss in identifying bacterial ghost to gap toe four [su Listeria produced therewith, in bacterial ghost has been confirmed that the remaining amount of DNA. Specifically, said in the embodiment <1-3>The method of adding hydrochloric acid or sodium hydroxide 60 minutes after Listeria bacteria in high purity, commercial extraction kit (iNtRON Biotechnology yarn, Korean) number using a number of the radiation [...] according to a protocol which genome DNA extracted from the. Then, 0. 5 g/Ml [...] bromide (ethidium bromide, EtBr) ethynyl loading objects to a gel electrophoresis was extracted extract including 1% direct recovery system of said embodiment. In addition, liver PCR embodiment performed quantitatively than to analyze the amount genome DNA, said extracted from a 1 micro l (1:100 dilution), each having a forward primer or reverse primer [table 3] bio-sequence listing for a 1 micro l, cyber green II QPCR 2 × master mix (Agilent Technology yarn, U.S.) and chuck 7 micro l blended, Stratagene Mx3000P embodiment [table 4] conditions such as liver PCR analyzer is provided which performs, for amplifying a genome DNA PCR amplified product remove the above amount of fluorometric embodiment performing liver has been confirmed. Nothing control hydrochloric or sodium hydroxide regulated finish bacteria Listeria to gap toe four [su O2, solvent controls include TE buffer solution processing deflection Listeria bacteria, positive controls include ammonium sulfate processing using Listeria bacteria, said method identical to embodiment conducting liver PCR analysis. Said forward primer and reverse primer comprises a recombinant expression vector of Listeria to gap toe four [su 16S rRNA portion designed specifically a gene amplification method are disclosed. As a result, as shown in the 6 also 5 and also, hydroxide or hydrochloric acid 60 minutes processing bacteria are simultaneously output of anger [thu loom ghost DNA has been confirmed (5 also). The compared, when quantitative analysis to liver PCR embodiment, sodium hydroxide when a trace amount of DNA is amplified in a location trace amounts of carbon monoxide Listeria to gap toe four [su while confirming the presence of bacterial ghost DNA, DNA amplification as well as hydrochloric acid as an amorphous when not processing, TE buffer solution has an Ct lower than has been confirmed (6 also). In this way, the present invention through hydrochloride or sodium hydroxide is processed by the genome DNA and mammalian proteins where the number prepared by the bacterial ghost, cell wall only held by this formate, in particular sodium hydroxide hydrochloric acid bacteria prepared by the number more low levels of genome DNA processing than the amount remaining ghost, foreign antigen as used for bacterial ghosts [...] scene or carrier may have negative effects of safety has been confirmed. <1-3> Bacteria Ghost The topography of the surface analysis identifying In the present invention number [...] foreign antigen such as a bacteria or bacterial ghost is effective as ghost scene produced therewith to determine whether used in order to identify, bacterial ghosts surface by observation under a scanning electron microscope (Scanning Electron Mircroscopy, SEM) conducting morphogenetic activity analysis. Specifically, said in the embodiment <1-3>The method of adding hydrochloric acid or sodium hydroxide 60 minutes after Listeria bacterial ghosts in high purity, 2. 5% chitosan non-suspended in PBS solution (glutaraldehyde) after adding 4 °C and fixed during time 2, cleaning in solutions was the same. Then, 1% preparing method of osmium (osmium tetroxide, OsO4 ) Solution in 1 bacterial ghost separated 4 °C. 5 fixed during time after, dehydrating a bacterial ghost was the dilution step into ethanol. Dehydrated bacterial ghost may include liquefied carbon dioxide drying sample prepared using electrolytic gold (gold) polaron high-a resolution sputter coating, S-a 4800 FESEM [...] bacterial ghosts observed surface scanning electron microscope. A method identical to the locus nothing control controls include Listeria performed SEM observed. As a result, as shown in Figure 7, compared to controls nothing control, hydrochloric acid processing hole sized tunnel structure on the bacterial ghost cells (arrows) in the apparent wall dissolved the, outer membrane form having a structure somewhat dissolved has been confirmed (also 7A). The compared, and slightly less than the size of the tunnel dissolved sodium hydroxide processing device, in the form of hydrochloric acid so as to surround the pathogenic effects related to processing compared to remain in complete form (also 7B) has been confirmed. The present invention relates to gram-positive bacterial ghosts prepared by hydrochloric acid treatment and to a method for preparing the same, and more particularly, the gram-positive bacillus bacterial ghosts prepared according to the method of the present invention can be effectively formed as bacterial ghosts when treated and cultured with a minimal inhibitory concentration (MIC) of hydrochloric acid capable of inhibiting the production of live bacteria colonies of gram-positive bacillus. Also, since the formed bacterial ghosts are configured so that the shape of the envelope of a cell is intactly preserved and proteins or DNAs in the cytoplasm do not remain, there is little risk of side effects such as secondary infections due to proliferation, when administered into a body. Accordingly, the bacterial ghosts of gram-positive bacillus prepared according to the method of the present invention can be effectively used as a vaccine or a foreign antigen carrier for prevention or treatment of gram-positive bacillus infectious diseases. COPYRIGHT KIPO 2017 I) incubating perpendicularly installed gram-positive bacteria; ii) said step i) are cultured in harvested from the gram to yield; iii) said step ii) processing to form a bacterial ghost gram negative and positive bacteria obtained in hydrochloric acid; and iv) said step iii) obtain including ghosts formed in bacteria, a gram-positive bacterial ghosts number bath method. According to Claim 1, said step i) of gram-positive bacteria [...] ( According to Claim 1, said step iii) treatment of the hydrochloric acid concentration (MIC) characterized in that the treatment with the minimum number billion, a gram-positive bacterial ghosts number bath method. According to Claim 3, said minimum number concentration 6 billion. Characterized in a 25 mg/Ml, a gram-positive bacterial ghosts number bath method. According to Claim 1, said step iii) 60 minutes to 10 characterized in that the treatment of the hydrochloric acid processing, a gram-positive bacterial ghosts number bath method. According to Claim 1, said step iii) 20 to 30 °C characterized in the treatment of the hydrochloric acid processing, a gram-positive bacterial ghosts number bath method. The number produced therewith according to Claim 1 method, a gram-positive bacterial ghost. According to Claim 7 active ingredient including a gram-positive bacterial ghost, vaccine composition for prevention and treatment of gram-positive infections. According to Claim 8, said [...] gram-positive bacteria ( According to Claim 8, said foreign antigen or vaccine compositions characterized as including the ghost scene [...] carrier bacteria, gram vaccine composition for prevention and treatment of infections. In vitro No. Chemical number processing concentration (mg/Ml) Hydrochloric acid (HCl) Sodium hydroxide (NaOH) Ammonium sulfate (NH4 )2 SO4 Calcium chloride CaCl2 0 (nothing control controls) - - - - 1 50 50 500 500 2 25 25 250 250 3 12. 5 12. 5 125 125 4 6. 25 6. 25 62. 5 62. 5 5 3. 125 3. 125 31. 25 31. 25 6 1. 5625 1. 5625 15. 625 15. 625 7 0. 78125 0. 78125 7. 8125 7. 8125 8 0. 390625 0. 390625 3. 90625 3. 90625 9 0. 1953125 0. 1953125 1. 953125 1. 953125 In vitro No. Hydrochloric acid processing group Sodium hydroxide processing group Hydrochloric acid concentration (mg/Ml) Motility dilution drainage Sodium hydroxide concentration (mg/Ml) Motility dilution drainage 0 - 10-3 - 10-3 4 6. 25 100 6. 25 100 5 3. 125 10-1 3. 125 100 Primer name Sequence A forward primer 5 'a-GGAATTCCACGTGTAGCGGTGAAAT-a 3' Reverse primer 5 'a-GACTACCAGGGTATCTAATCCTGTTTG-a 3' Temperature Time Repeat (cycle) 95 °C 10 minutes 1 times 95 °C 10 seconds 40 times 55 °C 10 seconds 72 °C 30 seconds
