CN1590405A - White candidas transcription factor gene and its use - Google Patents
White candidas transcription factor gene and its use Download PDFInfo
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Abstract
A Candida albicans transcription factor gene CaFLO8, its coding protein CaFlo8, and the application of said protein and its coding sequence are disclosed. Said protein CaFlo8 is an important toxic factor, taking part in the growth and pathogenic procedure of candida albicans.
Description
Technical field
The present invention relates to Candida albicans mycelial growth genes involved and Disease-causing gene.Particularly, relate to and from the genomic library of Candida albicans, be cloned into Candida albicans transcription factor gene CaFLO8 and uses thereof.The product C aFlo8 albumen of this genes encoding is important virulence factor, participates in the mycelial growth and the pathogenic course of Candida albicans.
Background technology
Candida albicans (Candida albicans) is a kind of a kind of opportunistic human body cause illness fungi that is separated to clinically, special in the patient of immune down regulation, as the organ transplantation patient, HIV patient etc., can cause superficial part and the infection of deep system widely, infection site comprises the oral cavity, and vagina etc. cause white mouth, vaginitis etc., also can invade epidermis and endotheliocyte and enter blood and arrive internal organs, as kidney, brain etc., cause septicemia, seriously can cause death (Odds, F.C.1994.J.Am.Acad, Dermatol.31:S2-S5.).
In recent years, Candida albicans has become No. second fungi killer that China is only second to red tinea disease, and oidiomycetic research has important medical and biological significance to white.
At present thorough not enough to the research of Candida albicans mechanism of causing a disease, it is pathogenic relevant with it that many factors are considered to, as adhesive capacity to epidermic cell, the proteolytic enzyme that secretion produces, colonial morphology and thalline are to (the Cutler JE.Annu Rev Microbiol such as conversion of mycelia form, 1991,45:187-218; KobayashiSC, Cutler JE.Microbiol.1998,6:92-94; Odds FC.J Am AcadDermatol, 1994,31 (pt2): s2-5). Candida albicans can exist with thalline (yeast form), mycelia, (hyphae), three kinds of forms of pseudohypha (pseudohyphae).Thalli morphology is one elliptical erythrocyte; The pseudohypha form can be thought the cell that a string form prolongs, but visible the hanging in separation site between cell contracted; The cell of mycelia form is very long, does not hang and contract in the separated place between the cell, and separation is not a complete closed.The thalline of Candida albicans is closely related to modality and its pathogenicity bo of mycelia, it is generally acknowledged that the easier adhesion of the Candida albicans of mycelia state and intrusion epithelium and endothelial layer increase the weight of to infect.The Candida albicans modality is subjected to the (Odds that induces of a lot of environmental factorss, F.C.1985. Morphogenesis in Candidaalbicans.Crit Rev Microbiol.12:45-93), comprise pH value, temperature, serum, N-acetyl ceramide, L-proline(Pro), experimental results show that pathogenic can the decline much or (the Lo that disappears of bacterial strain of mycelial growth defective, H.J.et al, Cell 90:939-949).
The amphiploid yeast saccharomyces cerevisiae can take place from the conversion of thalli morphology to the pseudohypha form when nitrogenous source lacks; Intussusception can take place in the monoploid yeast saccharomyces cerevisiae when carbon source lacks.The process of these metamorphosis comprises MAPK approach and cAMP/PKA approach by the mediation of many barss transduction pathway.Yeast saccharomyces cerevisiae and Candida albicans have certain similarity in the cell vital process, by the filamentous growth defective of complementary yeast saccharomyces cerevisiae mutant strain, obtain a series of and homologous gene each cascade component of yeast saccharomyces cerevisiae MAPK approach.CPH1 (STE12) (Liu H, Kohler J, Fink GR.Sience, 1994,266:1723-26, HST7 (STE7), CST20 (STE20) (Kohler JR, Fink GR.Proc Natl Acad Sci USA, 1996,93:13223-28 for example; Leberer E, Harcus D, Broadbent ID, Clark KL, et al.Proc Natl Acad Sci USA, 1996,93:13217-22), these genes knock out the defective that can cause Candida albicans mycelial growth on some solid medium.The modality of Candida albicans is by the mediation of many barss transduction pathway, and wherein MAPK and cAMP/PKA approach play important regulation.The transcription factor PHD1 homology of EFG1 and yeast saccharomyces cerevisiae, its coded product are transcription factors with HLH structural domain.EFG1 may be work in the cAMP/PKA approach (Sonneborn A, Bockmuhl DP, Gerads M, Kurpanek K, Sanglard D, ErnstJF.Mol Microbiol 2000 Jan; 35 (2): 386-96).The cAMP approach of the MAPK approach of Cph1 mediation and Efg1 mediation has obtained more research, Ras1 then work in the upstream of these two approach (FengQ, Summers E, Guo B, Fink G.J.Bacteriol.181:6339-6346).The bHLH transcription factor Cph2 of the homologous gene CaTEC1 of yeast saccharomyces cerevisiae TEC1 and Myc subfamily forms the influence that also has in various degree to the oidiomycetic mycelia of white, and CaTEC1 transcribe the regulation and control (SchweizerA that is subjected to Efg1 and Cph2, Rupp S, Taylor BN, Rollinghoff M, Schroppel K.Mol Microbiol 2000,38:435-445; Lane S, Zhou S, Pan T, Dai Q, Liu H, Mol Cell Biol 2001,21:6418-6428).Negative regulatory factor Tup1 and Rfg1 or Nrg1 suppress formation (the Kadosh D et al.Mol.Cell.Biol.21:2496-2505 of mycelia; Braun BR et al EMBO J.20:4753-4761.).
In sum, the disease that causes in view of Candida albicans gets more and more, so this area presses for exploitation and pathogenic relevant gene of Candida albicans and albumen.
Summary of the invention
The purpose of this invention is to provide a kind of new Candida albicans transcription factor gene and proteins encoded thereof that participates in the Candida albicans pathogenic effects.
Another object of the present invention provides the purposes of this polypeptide and encoding sequence thereof.
In a first aspect of the present invention, a kind of isolating Candida albicans CaFLO8 polypeptide is provided, this polypeptide is selected from down group:
(a) has the polypeptide of SEQ ID NO:2 aminoacid sequence;
(b) replacement, disappearance or the interpolation of SEQ ID NO:2 aminoacid sequence through one or more (1-15, preferably 1-10) amino-acid residue formed, and have transcriptional activation function by (a) polypeptides derived.Preferably, described transcriptional activation function refers to activate ECE1, and HWP1 or ALS1 transcribe.
In another preference, described polypeptide has the aminoacid sequence shown in the SEQ ID NO:2.
In a second aspect of the present invention, a kind of isolating polynucleotide are provided, it comprises a nucleotide sequence, and this nucleotide sequence is selected from down group:
(a) polynucleotide of code book invention aforementioned polypeptides;
(b) with polynucleotide (a) complementary polynucleotide.
In another preference, described polynucleotide encoding has the polypeptide of aminoacid sequence shown in the SEQ ID NO:2.
In another preference, the sequence of described polynucleotide has 298-2748 position among the SEQ ID NO:1.
In a third aspect of the present invention, a kind of carrier is provided, it contains the polynucleotide of the above-mentioned polynucleotide of the present invention, and a kind of host cell, and it contains above-mentioned carrier.
In fourth aspect present invention, provide a kind of can with Candida albicans CaFLO8 protein-specific bonded antibody of the present invention.
In a third aspect of the present invention, provide the carrier that contains polynucleotide of the present invention.
In a fourth aspect of the present invention, provide a kind of can with Candida albicans CaFlo8 protein-specific bonded antibody of the present invention.
In a fifth aspect of the present invention, preparation CaFlo8 is provided proteic method, this method comprises: (a) under expression condition, cultivate the above-mentioned host cell that is transformed or transduce; (b) from culture, isolate CaFlo8 albumen.
In a sixth aspect of the present invention, the nucleic acid molecule that can be used for detecting is provided, it contains a successive 15-3428 Nucleotide in the above-mentioned polynucleotide.
In a seventh aspect of the present invention, provide and whether had the proteic method of CaFlo8 in the test sample, it comprises: sample is contacted with the proteic specific antibody of CaFlo8, observe whether form antibody complex, formed antibody complex and just represented to exist in the sample CaFlo8 albumen.
In a eighth aspect of the present invention, provide the purposes of polypeptide of the present invention and encoding sequence.Polypeptide for example of the present invention can be used to screen the agonist that promotes the CaFlo8 polypeptide active, and perhaps screening suppresses the antagonist of CaFlo8 polypeptide active.The proteic encoding sequence of CaFlo8 of the present invention or its fragment can be used as primer and be used for pcr amplification reaction, perhaps are used for hybridization as probe, perhaps are used to make gene chip or microarray.
Description of drawings
Fig. 1 has shown the nucleotide sequence of Candida albicans CaFLO8 gene.
The reading frame nucleotide sequence that Fig. 2 has shown Candida albicans CaFLO8 gene is its deduced amino acid.
Fig. 3 has shown Candida albicans CaFlo8 and other proteic N end homologys relatively.
Fig. 4 has shown the strategy of CaFLO8 gene knockout.
Fig. 5 has shown the influence to the Candida albicans speed of growth of knocking out of CaFLO8 gene, and culture condition is the YPD solid medium, 25 ℃ or 42 ℃, and 3 days.
Fig. 6 has shown that the blocking-up Candida albicans mycelia that knocks out of CaFLO8 gene forms.Solid serum and Lee ' s substratum, 37 ℃ of cultivations.Liquid YPD adds 10% serum and Lee ' s substratum, 37 ℃ of cultivations.
Fig. 7 has shown the influence to the Candida albicans virulence of knocking out of CaFLO8 gene, the infection experiment of mouse system, and High is 1 * 10
7/ ml high-concentration bacterial liquid; Low is 1 * 10
6/ ml lower concentration bacterium liquid, every mouse tail vein injection 100 μ l bacterium liquid.
Fig. 8 has shown the regulation and control of CaFLO8 gene pairs mycelia and toxicity genes involved, carries out the Northern hybridization analysis with HWP1, ECE1, ALS1, CaFLO8 and ACT1 as probe.
Embodiment
Flo8 is an activating transcription factor, and growth of yeast saccharomyces cerevisiae pseudohypha and intussusception are played an important role, and the strain of flo8 disappearance can not be carried out intussusception, can not form pseudohypha.The flo8 disappearance causes the defective of yeast saccharomyces cerevisiae throwing out (flocculation).PKA may work by the Flo8 factor.
The inventor is through extensive and deep research, the chromogene storehouse of Candida albicans is changed over to the flo8 disappearance strain of yeast saccharomyces cerevisiae, the principle that utilization has complementary functions, screening obtains the homologous gene of FLO8, called after CaFLO8 (SEQ ID NO:1), its open reading frame contains 3428 Nucleotide, 817 the amino acid whose protein (SEQ ID NO:2) of encoding.The expression of CaFLO8 and knock out the mycelia that influences Candida albicans and form, the strain of caflo8 disappearance does not have toxicity to mouse.Therefore CaFLO8 is an important Disease-causing gene.
Test shows that (1) CaFlo8 can lack caused pseudohypha growth defect and intussusception defective by the complementary yeast saccharomyces cerevisiae Flo8 of part.(2) knocking out of CaFLO8 is non-lethal, but the disappearance of CaFLO8 gene has been blocked the formation of Candida albicans mycelia really, and this shows that the CaFLO8 gene is required for the Candida albicans mycelial growth really.(3) find that by the infection experiment of mouse system the strain of CaFLO8 gene knockout does not have toxicity.(4) CaFLO8 gene regulating mycelia forms the expression of genes involved and virulence associated gene.Therefore, CaFlo8 albumen is virulence factor and activating transcription factor important in the Candida albicans, participates in the pathogenic course of Candida albicans.Finished the present invention on this basis.
In the present invention, term " CaFlo8 albumen ", " CaFlo8 polypeptide " or " Candida albicans CaFlo8 polypeptide " are used interchangeably, all refer to the to have Candida albicans CaFlo8 aminoacid sequence albumen or the polypeptide of (SEQ ID NO:2), and SEQ ID NO:2 aminoacid sequence formed through replacement, disappearance or the interpolation of one or more amino-acid residues, and have transcriptional activation function by (a) polypeptides derived.
As used herein, " isolating " is meant that material separates (if natural substance, primal environment promptly is a natural surroundings) from its primal environment.Do not have separation and purification as polynucleotide under the native state in the active somatic cell and polypeptide, but same polynucleotide or polypeptide as from native state with in other materials that exist separately, then for separation and purification.
As used herein, " isolating Candida albicans CaFlo8 albumen or polypeptide " is meant that Candida albicans CaFlo8 polypeptide is substantially free of natural relative other albumen, lipid, carbohydrate or other material.Those skilled in the art can use the purified technology of protein purifying Candida albicans CaFlo8 albumen of standard.Basically pure polypeptide can produce single master tape on non-reduced polyacrylamide gel.
In the present invention, term " Candida albicans CaFlo8 polypeptide " refers to have the SEQ ID NO.2 polypeptide of sequence of Candida albicans CaFlo8 protein-active.This term also comprises having and variant form Candida albicans CaFlo8 albumen identical function, SEQ ID NO.2 sequence.These variant forms comprise (but being not limited to): several (are generally 1-50, preferably 1-30, more preferably 1-20,1-10 best) amino acid whose disappearance, insertion and/or replacement, and add one or several at C-terminal and/or N-terminal and (be generally in 20, preferably being in 10, more preferably is in 5) amino acid.For example, in the art, when replacing, can not change proteinic function usually with the close or similar amino acid of performance.Again such as, add one or several amino acid at C-terminal and/or N-terminal and also can not change proteinic function usually.This term also comprises proteic active fragments of Candida albicans CaFlo8 and reactive derivative.
In the present invention, " Candida albicans CaFlo8 albumen conservative property variation polypeptide " refers to compare with the aminoacid sequence of SEQ ID NO:2, there are 10 at the most, preferably at the most 8, more preferably at the most 5,3 amino acid is replaced by similar performance or close amino acid and is formed polypeptide at the most best.These conservative property variation polypeptide preferably carry out the amino acid replacement according to table 1 and produce.
Table 1
| Initial residue | Representational replacement | The preferred replacement |
| Ala(A) | Val；Leu；Ile | Val |
| Arg(R) | Lys；Gln；Asn | Lys |
| Asn(N) | Gln；His；Lys；Arg | Gln |
| Asp(D) | Glu | Glu |
| Cys(C) | Ser | Ser |
| Gln(Q) | Asn | Asn |
| Glu(E) | Asp | Asp |
| Gly(G) | Pro；Ala | Ala |
| His(H) | Ash；Gln；Lys；Arg | Arg |
| Ile(I) | Leu；Val；Met；Ala；Phe | Leu |
| Leu(L) | Ile；Val；Met；Ala；Phe | Ile |
| Lys(K) | Arg；Gln；Asn | Arg |
| Met(M) | Leu；Phe；Ile | Leu |
| Phe(F) | Leu；Val；Ile；Ala；Tyr | Leu |
| Pro(P) | Ala | Ala |
| Ser(S) | Thr | Thr |
| Thr(T) | Ser | Ser |
| Trp(W) | Tyr；Phe | Tyr |
| Tyr(Y) | Trp；Phe；Thr；Ser | Phe |
| Val(V) | Ile；Leu；Met；Phe；Ala | Leu |
Polynucleotide of the present invention can be dna form or rna form.Dna form comprises the DNA of cDNA, genomic dna or synthetic.DNA can be strand or double-stranded.DNA can be coding strand or noncoding strand.The coding region sequence of encoding mature polypeptide can be identical with the coding region sequence shown in the SEQ ID NO:1 or the varient of degeneracy.As used herein, " varient of degeneracy " is meant that in the present invention coding has the protein of SEQ ID NO:2, but with the differentiated nucleotide sequence of coding region sequence shown in the SEQ ID NO:1.
The polynucleotide of the mature polypeptide of coding SEQ ID NO:2 comprise: the encoding sequence of an encoding mature polypeptide; The encoding sequence of mature polypeptide and various additional code sequence; Encoding sequence of mature polypeptide (with optional additional code sequence) and non-coding sequence.
The invention still further relates to and above-mentioned sequence hybridization and two sequences between have at least 50%, preferably at least 70%, more preferably at least 80%, the polynucleotide of at least 90% homogeny best.The present invention be more particularly directed under stringent condition and the interfertile polynucleotide of polynucleotide of the present invention.In the present invention, " stringent condition " is meant: (1) than hybridization under low ionic strength and the comparatively high temps and wash-out, as 0.2 * SSC, and 0.1%SDS, 60 ℃; Or (2) hybridization the time is added with denaturing agent, as 50% (v/v) methane amide, 0.1% calf serum/0.1%Ficoll, 42 ℃ etc.; Or (3) only at the homogeny between the two sequences at least more than 90%, be more preferably 95% and just hybridize when above.And the polypeptide of interfertile polynucleotide encoding has identical biological function and activity with the mature polypeptide shown in the SEQ ID NO:2.
The invention still further relates to nucleic acid fragment with above-mentioned sequence hybridization.As used herein, the length of " nucleic acid fragment " contains 15 Nucleotide at least, better is at least 30 Nucleotide, is more preferably at least 50 Nucleotide, preferably more than at least 100 Nucleotide.Nucleic acid fragment can be used for the amplification technique (as PCR) of nucleic acid to determine and/or the proteic polynucleotide of separation coding Candida albicans CaFlo8.
Candida albicans CaFlo8 Nucleotide full length sequence of the present invention or its fragment can obtain with the method for pcr amplification method, recombination method or synthetic usually.For the pcr amplification method, can be disclosed according to the present invention about nucleotide sequence, especially open reading frame sequence designs primer, and with commercially available dna library or by the prepared dna library of ordinary method well known by persons skilled in the art as template, amplification and must relevant sequence.When sequence is longer, usually needs to carry out twice or pcr amplification repeatedly, and then the fragment that each time amplifies is stitched together by proper order.
In case obtained relevant sequence, just can obtain relevant sequence in large quantity with recombination method.This normally is cloned into carrier with it, changes cell again over to, separates obtaining relevant sequence then from the host cell after the propagation by ordinary method.
In addition, also the method for available synthetic is synthesized relevant sequence, especially fragment length more in short-term.Usually, by first synthetic a plurality of small segments, and then connect and to obtain the very long fragment of sequence.
At present, can be fully obtain the dna sequence dna of code book invention albumen (or its fragment, or derivatives thereof) by chemosynthesis.This dna sequence dna can be introduced in various existing dna moleculars as known in the art (or as carrier) and the cell then.In addition, also can will suddenly change and introduce in the protein sequence of the present invention by chemosynthesis.
Use method (Saiki, the et al.Science1985 of round pcr DNA amplification/RNA; 230:1350-1354) be optimized for acquisition gene of the present invention.The primer that is used for PCR can suitably be selected according to sequence information of the present invention disclosed herein, and available ordinary method is synthetic.Available ordinary method is as the DNA/RNA fragment by gel electrophoresis separation and purifying amplification.
The present invention also relates to comprise the carrier of polynucleotide of the present invention, and the host cell that produces through genetically engineered with carrier of the present invention or Candida albicans CaFlo8 albumen coded sequence, and the method that produces polypeptide of the present invention through recombinant technology.
Recombinant DNA technology (Science, 1984 by routine; 224:1431), can utilize polymerized nucleoside acid sequence of the present invention to can be used to express or produce the CaFlo8 polypeptide of reorganization.In general following steps are arranged:
(1). with the polynucleotide (or varient) of coding Candida albicans CaFlo8 polypeptide of the present invention, or transform or the transduction proper host cell with the recombinant expression vector that contains these polynucleotide;
(2). the host cell of in suitable medium, cultivating;
(3). separation, protein purification from substratum or cell.
Among the present invention, Candida albicans CaFlo8 polynucleotide sequence can be inserted in the recombinant expression vector.Term " recombinant expression vector " refers to that bacterial plasmid well known in the art, phage, yeast plasmid, vegetable cell virus, mammalian cell virus are as adenovirus, retrovirus or other carriers.In a word, as long as can duplicate in host and stablize, any plasmid and carrier can be used.A key character of expression vector is to contain replication orgin, promotor, marker gene and translation controlling elements usually.
Method well-known to those having ordinary skill in the art can be used to make up and contains Candida albicans CaFlo8 DNA sequences encoding and suitable transcribing/the translate expression vector of control signal.These methods comprise extracorporeal recombinant DNA technology, DNA synthetic technology, the interior recombinant technology of body etc.Described dna sequence dna can effectively be connected on the suitable promotor in the expression vector, and is synthetic to instruct mRNA.The representative example of these promotors has: colibacillary lac or trp promotor; Lambda particles phage PL promotor; Eukaryotic promoter comprises LTRs and some other known may command gene expression promoter in protokaryon or eukaryotic cell or its virus of CMV immediate early promoter, HSV thymidine kinase promoter, early stage and late period SV40 promotor, retrovirus.Expression vector also comprises ribosome bind site and the transcription terminator that translation initiation is used.
Host cell can be a prokaryotic cell prokaryocyte, as bacterial cell; Or eukaryotic cell such as low, as yeast cell; Or higher eucaryotic cells, as mammalian cell.Representative example has: intestinal bacteria, streptomyces; The bacterial cell of Salmonella typhimurium; Fungal cell such as yeast; Vegetable cell; The insect cell of fruit bat S2 or Sf9; The zooblast of CHO, COS, 293 cells or Bowes melanoma cells etc.
Can carry out with routine techniques well known to those skilled in the art with the recombinant DNA transformed host cell.When the host was prokaryotic organism such as intestinal bacteria, the competent cell that can absorb DNA can be used CaCl in exponential growth after date results
2Method is handled, and used step is well-known in this area.Another kind method is to use MgCl
2If desired, transforming also the method for available electroporation carries out.When the host is an eukaryote, can select following DNA transfection method for use: coprecipitation of calcium phosphate method, conventional mechanical method such as microinjection, electroporation, liposome packing etc.
The transformant that obtains can be cultivated with ordinary method, expresses the polypeptide of coded by said gene of the present invention.According to used host cell, used substratum can be selected from various conventional substratum in the cultivation.Under the condition that is suitable for the host cell growth, cultivate.After host cell grows into suitable cell density, induce the promotor of selection with suitable method (as temperature transition or chemical induction), cell is cultivated for some time again.
The extracellular can be expressed or be secreted into to recombinant polypeptide in the above methods in cell or on cytolemma.If desired, can utilize its physics, the separating by various separation methods with other characteristic and the albumen of purification of Recombinant of chemistry.These methods are well-known to those skilled in the art.The example of these methods includes, but are not limited to: conventional renaturation handles, with protein precipitant handle (salt analysis method), centrifugal, the broken bacterium of infiltration, superly handle, the combination of super centrifugal, sieve chromatography (gel-filtration), adsorption chromatography, ion exchange chromatography, high performance liquid chromatography (HPLC) and other various liquid chromatography (LC) technology and these methods.
The Candida albicans CaFlo8 albumen or the polypeptide of reorganization are of use in many ways.These purposes include, but is not limited to: be used to screen and promote or to antibody, polypeptide or other part of anti-candida albicans CaFlo8 protein function.The peptide molecule that can suppress or stimulate Candida albicans CaFlo8 protein function that can be used for seeking therapeutic value with the reorganization Candida albicans CaFlo8 protein screening peptide library of expressing.
On the other hand, the present invention also comprises Candida albicans CaFlo8 DNA or the polypeptide of its fragment coding has specific polyclonal antibody and monoclonal antibody, especially monoclonal antibody.Here, " specificity " is meant that antibody capable is incorporated into Candida albicans CaFlo8 gene product or fragment.Preferably, refer to that those can combine with Candida albicans CaFlo8 gene product or fragment but nonrecognition and be incorporated into the antibody of other irrelevant antigen molecule.
The present invention not only comprises complete mono-clonal or polyclonal antibody, but also comprises having immunocompetent antibody fragment, as Fab ' or (Fab)
2Fragment; Heavy chain of antibody; Light chain of antibody; Genetically engineered strand Fv molecule; Or chimeric antibody.Antibody of the present invention can be prepared by the known various technology of those skilled in that art.For example, the Candida albicans CaFlo8 gene product of purifying or its have antigenic fragment, can be applied to animal to induce the generation of polyclonal antibody.Similarly, expressing Candida albicans CaFlo8 albumen or its has antigenic segmental cell and can be used to immune animal and produce antibody.Antibody of the present invention also can be monoclonal antibody.This type of monoclonal antibody can utilize hybridoma technology to prepare.
The proteic antibody of anti-candida albicans CaFlo8 can be used in the immunohistochemistry technology, detects the Candida albicans CaFlo8 albumen in the biopsy specimen.
Utilize albumen of the present invention,, can filter out with Candida albicans CaFlo8 albumen interactional material takes place, as acceptor, inhibitor, agonist or antagonist etc. by various conventional screening methods.
The invention still further relates to the diagnostic testing process of quantitative and detection and localization Candida albicans CaFlo8 protein level.These tests are known in the art.
Whether having the proteic method of Candida albicans CaFlo8 in a kind of detection test sample is to utilize the proteic specific antibody of Candida albicans CaFlo8 to detect, and it comprises: sample is contacted with Candida albicans CaFlo8 protein specific antibody; Observe whether form antibody complex, formed antibody complex and just represented to exist in the sample Candida albicans CaFlo8 albumen.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, people such as Sambrook for example, molecular cloning: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
Universal method:
Southern analyzes
Candida albicans gene group DNA method for extracting and Southern hybridization analysis method are according to document operation (Chen et al.J.Bacteriol.174:5624-56312.).Genomic dna PvuII (available from GIBCO company) complete degestion, agarose gel after electrophoresis is finished soaks 45min with sex change liquid and neutralizer respectively, and nylon membrane soaks into distilled water or 10 * SSC, puts up and changes the film platform, glue and intermembranous with the Parafilm sealing adds a 500g counterweight., serve as to change the transfer of film liquid to spend the night with 10 * SSC, rinsing in second day is dried crosslinked.Film after the crosslinked hybrid pipe of packing into adds 10ml prehybridization solution (6 * SSC, 5 * denhardt ' sReagent, 0.5%SDS, 100 μ g/ml milt DNA are in 42 ℃ of prehybridization 12h, and probe adds 42 ℃ of hybridization of 150 μ l (probe of about 1/3 mark) 10-16h at 100 ℃ of sex change 5min.0.1 * SSC, 0.1%SDS wash two to three times, each 40min takes out film, dries room temperature or-70 ℃ of compressing tablets.Probe mark is operated with the random primer labelling test kit of invitrogene and according to its explanation.The solution that will contain 25ng DNA is in 100 ℃ of heat denatured 5-10min, place ice bath, add Random Primers Buffer Mixture more successively, 2 μ l dCTP, 2 μ l dGTP, 2 μ l dTTP, 3 μ l[α-32P]-dATP (10 μ Ci/ μ l), add 1 μ l Klenow enzyme behind the mixing, in 25 ℃ of insulation 1h, with the centrifugal 4min of 3000rpm, collect centrifugate with Sephadex G-25 post, be the probe solution behind the purifying.Probe adds in the hybridization system after 100 ℃ of sex change 5min cooled on ice.
Northern analyzes
With the total RNA of hot phenol method extracting Candida albicans, and Northern hybridization analysis method reference literature (Greenberg, M.E.1987, Current protocols in molecular biology, vol.1.WileyInterscience, New York, N.Y.).Each bacterial strain is at YPD, is cultured to the logarithmic phase later stage in 30 ℃, is forwarded to YPD+10% serum, and perhaps Lee ' s substratum is 1, initial OD=0.05, and under relevant temperature, cultivate the fixed time, with the total RNA of hot phenol method extracting Candida albicans.20 μ l applied sample amounts comprise RNA sample 12 μ l (25 μ g), 4 μ l formaldehyde, 2 μ l, 10 * MOPS, 2 μ l sample-loading buffers, 65 ℃ of heating 10min, 0 ℃ of cooling 2min, centrifugal 5s, last sample.Electrophoresis carries out (87ml DEPC treating water, the cold slightly back of heating and melting adds 2.7ml 37% formaldehyde for 1g agarose, 10ml 10 * MOPS) containing on the sex change glue of formaldehyde.Deposition condition is 100mA, and 50-70V changes film equally after the 5-7h electrophoresis is finished with the capillary method, changes film liquid with 5 * SSC.After the commentaries on classics film spends the night, take out film, film is parched, UV-crosslinked with Bio-Ra GS Gene Linker C3 protocol, add the 10ml prehybridization solution, 65 ℃, be replaced by the 10ml hybridization solution behind the 2h, add 65 ℃ of hybridization of probe and spend the night, with 2 * SSC, 0.1%SDS washes film twice for 65 ℃, each 30min.The film that hybridization is good does not dry, and wraps-70 ℃ of compressing tablets in back with preservative film.Heavily film 0.5%SDS is washed in hybridization, and 90-100 ℃ of heating 5-10min, or recommend use 0.5%SDS according to Clontech takes out standbyly behind 90-100 ℃ of its naturally cooling of heating 10min relief 10min, as if a usefulness immediately not, wrap with preservative film and to be placed on-20 ℃ of preservations.
The conversion of Candida albicans and yeast saccharomyces cerevisiae
Prepare PEG/LiAc solution (pH 7.5) 10ml; In 1.5mI Eppendof pipe, add plasmid (if change yeast saccharomyces cerevisiae, plasmid 0.1 μ g is if change Candida albicans, the about 5 μ g of linearization plasmid) successively, 10 μ l10mg/ml milt DNA, mixing; Add 0.1ml competent cell and 0.6ml PEG/LiAc, the votex mixing; 30 ℃ of 200rpm cultivate 30min; Add 70 μ l DMSO, mixings gently; 42 ℃ of thermal shocking 15min, during shake up gently frequently; Ice bath～2min: high speed centrifugation 15s sops up supernatant, with 0.2ml TE re-suspended cell; Coat on the suitable nutrition screening SD flat board.
The infection experiment of mouse system
Is experimental subjects with body weight at 16-18g ICR male mice, by tail vein injection 100 each bacterial strain of μ l Candida albicans, and cultivates observation 25 days.Two concentration of each injection height of each bacterial strain, high-concentration bacterial liquid is 5 * 10
7Cell/ml, lower concentration bacterium liquid is 5 * 10
6Cell/ml.
The extracting of Candida albicans total protein, immunoprecipitation and Western Blot detect.
Albicans strain is cultured to OD=1 in YPD, collecting cell is resuspended in 400 μ l yeast cell lysates with after the 400 μ l yeast cell lysates washing 3 times, add 0.5mm pearl 0.2g, in 0 ℃, on Minibead-beater, vibrate twice, each 30 seconds with 4800rpm.The centrifugal 15min of 12000rpm draws supernatant, centrifugal again 15min, and the collection supernatant is a cell lysate.Get 500 μ l cell lysates, add anti-FLAG monoclonal antibody (Sigma) (2 μ g/ reaction), at 4 ℃ of vibration 1h.The centrifugal 2min of 12000rpm adds 50% Protein G-agarose (Roche MolecularBiochemicals) suspension that 30 μ l cross with yeast cell lysate balance in supernatant, 4 ℃ of vibration 2h.Immunoprecipitate is given a baby a bath on the third day after its birth inferior with IP damping fluid (20mM Tris-HCl pH 7.5,2mM EGTA, 150mM NaCl, 1%NP-40,1mM DTT).Be resuspended in then in 1 * protein electrophorese sample-loading buffer, carry out 10% SDS-PAGE electrophoresis.With wet method protein band is transferred on the nitrocellulose filter, with the TBS-T that contains 5% skim-milk (10mM Tris-HCl (pH 7.5), 150mMNaCl, 0.1%Tween-20) solution closing membrane 2-3 hour, 4 ℃ of combinations of anti-FLAG (Sigma) (antibody dilution is 1: 1000) are spent the night, wash film four times with the TBS-T room temperature, each 15 minutes.Resist in room temperature in conjunction with 2 hours with two, wash film four times with TBS-T again, each 15 minutes.Exhaust excess liquid with thieving paper, use the ECL reagent colour development.
Embodiment 1:
The clone of CaFLO8 gene
Will be with ordinary method (Liu H, K hler J, Fink GR.Science, 1994,266:1723-1726) the chromogene storehouse of the Candida albicans of Gou Jianing changes yeast saccharomyces cerevisiae monoploid flo8 disappearance strain (Liu H over to, Styles CA, Fink GR.Genetics, 1996,144:967-978, and can be available from Yeast Genetics StockCulture Center of the American Type Culture Collection), on the SD-ura flat board, grow, obtain 30,000 bacterium colonies.Water washes these transformed bacterias, and the bacterium colony that is not rinsed is chosen as the clone of complementary intussusception defective, obtains 75 positive colonies altogether.
Extracting yeast plasmid by the following method: inoculate single bacterium colony in 5ml SD ura-liquid nutrient medium, 30 ℃ of shaking culture 20h, the centrifugal 3min of room temperature 5000r/min, abandon supernatant, add 200 μ l yeast lysate (2%Triton X-100,1%SDS, 100mmol/L NaCl, 10mmol/L Tris-HCl pH 8.0,1mM EDTA), resuspended yeast vibrates, add 0.3mg pickling glass pearl (425～600 μ m), 200 μ l phenol: chloroform: primary isoamyl alcohol (25: 24: 1), the 5min that fully vibrates, liquid nitrogen freezes 10min, room temperature is melted, the 5min that fully vibrates again, the centrifugal 10min of room temperature 12000r/min is transferred to new 1.5ml centrifuge tube with supernatant liquor, add 20 μ l 3mol/L NaAc, 500 μ l dehydrated alcohols ,-70 ℃ of frozen 1h, 4 ℃ of centrifugal 10min of 12500r/min, abandon supernatant, 70% washing with alcohol precipitation is drained, and adds 20 μ l TE (pH 8.0) dissolving.
Plasmid that will extracting goes out from these bacterium colonies, electricity changes intestinal bacteria DH12S over to, will carry out restriction analysis by the plasmid of extractive purifying from intestinal bacteria.Behind the restriction enzyme mapping preliminary classification, again plasmid is changed again over to the strain of monoploid and amphiploid flo8 disappearance respectively, observe the intussusception and the filamentous growth phenotype of bacterium colony, find that most of plasmid can both complementary to some extent flo8 intussusception and filamentous growth defective.With No. 56 plasmid order-checkings, and analyze comparison by tBlast n and the Blast p of NCBI, obtain inserting segmental complete sequence, 5 ' and 3 ' the end non-coding region that comprises open reading frame, total length has 3428 Nucleotide (Fig. 1 and SEQ IDNO:1), its ORF is positioned at the 298-2748 position, the albumen (Fig. 2 and SEQ ID NO:2) of coding total length 817aa.Because the N of its N end protein sequence and yeast saccharomyces cerevisiae Flo8 factor end has homology, and the flo8 defective strain of the yeast saccharomyces cerevisiae that can have complementary functions again, be this unnamed gene CaFLO8 therefore, the albumen of CaFLO8 genes encoding is called CaFlo8.We deposit gene order in the GenBank database, and sequence number is AF414113 (this sequence is underground before the application submits to).
Embodiment 2
Candida albicans CaFLO8 gene can the complementary ScFlo8 functional defect of part
CaFlo8 can complementary ScFlo8 lacks the defective of caused pseudohypha growth and intussusception, so Candida albicans CaFLO8 gene and yeast saccharomyces cerevisiae FLO8 have certain similarity at sequential structure and function aspects.
The CaFLO8 total length is 3428bp (SEQ ID NO:1), is inferred the albumen (SEQ ID NO:2) of a 817aa of its coding by sequence.This albumen is highly hydrophilic, and middle part (126-396) is rich in glutamine, and C holds the acidic region of the 130aa that has an appointment.The 62aa (32-94aa) and Flo8 (Sacchromyces cerevisiae) of Caflo8 N end, Leunig (Arabidopsis thaliana), Leuni (Oryza sativa) and putative WD-40 repeat protein (Oryza sativa), the N terminal sequence of putative flower development regulator LEUNI protein (Oryza sativa) has homology (Fig. 3).
The structural similitude of CaFlO8 and Flo8 and Leunig, glutamine enrichment region (Q-rich) zone (the 112-395aa) (Conner that all has N end LUFS structural domain (LUFS domain) and middle part, J.and Liu, Z.Proc.Natl.Acad.Sci.U.S.A.2000,97,23,12902-12907).
The N end of finding CaFlo8 with the SMART inquiry is LisH (LIS1 homology) structural domain (31-63aa), and the coincidence of the first half of LisH structural domain and LUFS structural domain, and there is spiral (coiledcoil) structural domain of spiral at the middle part.Some eukaryotic cell intracellular proteins that relate to microtubule kinetics, cell migration and chromosome segregation contain Lis H structural domain.The LisH structural domain has conservative protein binding function, and the LisH structural domain acts on by mediating dimerization or being attached on born of the same parents' internally-powered albumen (dynein) heavy chain or the microtubule the dynamic (dynamical) regulation and control of microtubule according to inferring.The secondary structure of LisH structural domain is made up of two α spirals, and coiled coil structural domain is considered to relevant with the multimerization of subunit.Therefore, N end LUFS structural domain may be relevant with the function of the CaFlo8 factor, and perhaps this effect relates to proteic multimerization.
Embodiment 4
Knocking out of Candida albicans CaFLO8 gene
In order to study the function of Candida albicans CaFLO8 gene, in the present embodiment, utilize principle of homologous recombination to knock out the CaFLO8 gene at Candida albicans.
The coding region of the one section about 0.9kb in centre of CaFLO8 gene is replaced by the hisG-URA3-hisG fragment, the CaFLO8 gene that changes over to after the linearizing on bacterial strain and the karyomit(e) carries out homologous recombination (Fig. 4), conversion obtains the strain of CaFLO8 gene knockout through two-wheeled, and the disappearance strain is identified with the method for Southern hybridization.
The result:
1.CaFLO8 gene knock out the influence that the oidiomycetic growth of white and mycelia are formed
Knocking out of CaFLO8 is non-lethal, and the disappearance strain can both be grown under the condition of low temperature (22 ℃) and high temperature (42 ℃), but influential to the oidiomycetic speed of growth of white.Under several growth conditionss (25 ℃, 30 ℃, 37 ℃, 42 ℃), the caflo8 disappearance strain speed of growth is all slower.CaFLO8 is imported again, then can reply the defective of bacterium colony slow growth, illustrate that this growth defect is (Fig. 5) that the disappearance by the CaFLO8 gene causes.
In multiple mycelia inducing culture, wild type strain can form long mycelia, and the mycelia formation of Candida albicans is all blocked in the strain of caflo8 disappearance under various mycelia inductive conditions, even add at YPD under the inductive condition of 10% calf serum, can not form mycelia.In solid medium, the strain of caflo8 disappearance all is to form slick bacterium colony, and wild type strain can form long mycelia.CaFLO8 is imported again, and the mycelia that then can reply Candida albicans forms, and illustrates that the CaFLO8 gene is Candida albicans mycelial growth required (Fig. 6) really.
2.CaFLO8 gene knock out influence to the Candida albicans virulence
Detect of the influence of the disappearance of CaFLO8 gene by the infection experiment of mouse system to the Candida albicans virulence.With the positive contrast of wild type strain, detected the toxicity of caflo8 disappearance strain.Wild type strain is all dead in 4 days under the high density condition, and also all death in 10 days under low consistency conditions.The strain of caflo8 disappearance does not then have toxicity, does not have death in experiment, and the mouse after the injection still all survived and weight increase (Fig. 7) after 30 days.CaFLO8 genetically deficient is consistent with it to the influence that mycelia forms to the influence of Candida albicans virulence.
3.CaFLO8 the gene regulating mycelia forms the expression of genes involved and virulence associated gene
The mycelial growth that knocks out the blocking-up Candida albicans of CaFLO8 gene, the strain of proof disappearance does not have toxicity in the mouse toxicity experiment, and therefore the method with Northern hybridization detects it to mycelia and toxicity genes involved ECE1, HWP1, the transcriptional control of ALS1.ECE1 (Birse et al.Infect.Immun.1993,61:3648-3655) and HWP1 (Staab et al.J.Biol.Chem.1996 271:6298-6305) be the mycelia specific gene, great expression under the mycelia inductive condition; ALS1 and Candida albicans are to relevant (the Yue F et al of epithelial adhesion.Mol?Microl?2002?44：61-72)。
The result shows that ECE1 has been blocked in knocking out under the mycelia inductive condition of CaFLO8 gene, HWP1, and the expression of ALS1 (Fig. 8), the influence that the generation of CaFLO8 gene pairs mycelia is described is by regulating and control to work to the mycelia Expression of Related Genes.Prompting CaFLO8 has transcriptional activation function.
Embodiment 4
The expression of CaFLO8
The CaFLO8 encoder block obtains from the wild type strain pcr amplification with pfu archaeal dna polymerase (available from Promega company), cuts 1 hour with BamH I and 37 ℃ of enzymes of Sph I (available from GIBCO company).Candida albicans high-expression vector pFLAG-Act1 (Umeyama T et al.Yeast 2002; 19:611-618) handled back 1 hour for 37 ℃ with BamH I and Sph I, be connected 1 hour with the section of PCR enzyme is disconnected for 22-26 ℃ with T4 dna ligase (available from GIBCO company), transformed into escherichia coli, the picking mono-clonal, the extraction plasmid is cut evaluation with BamH I and Sph I enzyme and is obtained the CaFLO8 expression plasmid.The used primer of amplification CaFLO8 encoder block is primer 1:5 '-CTGGGATTCATGAATCATAAACAAGTACTA-3 ' (SEQ ID NO:3) and primer 2: 5 '-CTGCGCATGCATCGCCATTTTCAATTGGATC-3 ' (SEQ ID NO:4).
PFLAG-Act1-CaFLO8 is cut with the NcoI enzyme, transform Candida albicans, with glass bead method extracting yeast total protein, with antibody (available from the sigma company) detection of Anti-FLAG, the result has obtained the FLAG-CaFlo8 expressing protein that molecular weight is about 90Kda, and molecular weight conforms to predictor.
All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Sequence table
＜110〉Shanghai Inst. of Life Science, CAS
＜120〉Candida albicans transcription factor gene and uses thereof
<130>034756
<160>4
<170>PatentIn?version?3.1
<210>1
<211>3428
<212>DNA
＜213〉Candida albicans (Candida albicans)
<220>
<221>CDS
<222>(298)..(2748)
<223>
<400>1
tcacactggc?tatcttaccc?acttttatca?acgaacagta?tatcaaactg?cacttttttt?????60
atacaaattt?gaaaagttca?acaaaaaaca?acaatattac?tttatatata?tccatttgac????120
aattttcata?catttttttc?attaaatctg?ctatattttg?ttgcttttat?taatttattg????180
gctttttatt?ttgttttggt?ttgttttgtt?tcatttgttt?tattttcctt?taaagaaatt????240
tcattattat?cttcatatac?atacatatat?aactcataac?tacaattatt?gctggca???????297
atg?aat?cat?aaa?caa?gta?cta?cca?gaa?aat?gaa?ccg?ata?tca?aca?aca??????345
Met?Asn?His?Lys?Gln?Val?Leu?Pro?Glu?Asn?Glu?Pro?Ile?Ser?Thr?Thr
1???????????????5???????????????????10??????????????????15
aca?gca?act?cca?tca?tca?tcc?aat?act?atg?gtt?ccc?aac?aca?act?aaa??????393
Thr?Ala?Thr?Pro?Ser?Ser?Ser?Asn?Thr?Met?Val?Pro?Asn?Thr?Thr?Lys
20??????????????????25??????????????????30
cag?gtt?tta?aac?tcg?ttg?att?ttg?gat?ttt?ttg?gtc?aaa?cat?caa?ttt??????441
Gln?Val?Leu?Asn?Ser?Leu?Ile?Leu?Asp?Phe?Leu?Val?Lys?His?Gln?Phe
35??????????????????40??????????????????45
caa?gat?aca?gca?aaa?gca?ttt?tct?aaa?gaa?agt?ccc?aat?ttg?cct?tct??????489
Gln?Asp?Thr?Ala?Lys?Ala?Phe?Ser?Lys?Glu?Ser?Pro?Asn?Leu?Pro?Ser
50??????????????????55??????????????????60
att?cct?cct?ttg?atg?gat?tgt?tct?caa?gga?ttt?tta?ttg?gaa?tgg?tgg??????537
Ile?Pro?Pro?Leu?Met?Asp?Cys?Ser?Gln?Gly?Phe?Leu?Leu?Glu?Trp?Trp
65??????????????????70??????????????????75??????????????????80
caa?gtt?ttc?ttt?gat?tta?ttt?caa?gtc?aga?tat?gga?gac?ggt?aac?agt?????585
Gln?Val?Phe?Phe?Asp?Leu?Phe?Gln?Val?Arg?Tyr?Gly?Asp?Gly?Asn?Ser
85??????????????????90??????????????????95
aat?aat?aac?cct?aac?aac?aag?ctt?tat?cat?gat?tat?ctc?aga?gtc?caa?????633
Asn?Asn?Asn?Pro?Asn?Asn?Lys?Leu?Tyr?His?Asp?Tyr?Leu?Arg?Val?Gln
100?????????????????105?????????????????110
gaa?act?caa?aaa?cat?ctt?ttc?agt?caa?ctt?cct?ctt?ata?cag?cag?cag?????681
Glu?Thr?Gln?Lys?His?Leu?Phe?Ser?Gln?Leu?Pro?Leu?Ile?Gln?Gln?Gln
115?????????????????120?????????????????125
caa?caa?caa?caa?cat?cac?ttt?caa?caa?caa?cag?caa?caa?caa?ggg?caa?????729
Gln?Gln?Gln?Gln?His?His?Phe?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gly?Gln
130?????????????????135?????????????????140
cag?gga?cag?cct?ttc?ctg?cag?caa?cag?caa?aga?gga?atc?ggt?gtt?gct?????777
Gln?Gly?Gln?Pro?Phe?Leu?Gln?Gln?Gln?Gln?Arg?Gly?Ile?Gly?Val?Ala
145?????????????????150?????????????????155?????????????????160
agt?ggt?atg?caa?aat?caa?caa?cat?caa?ttt?gcc?cca?cag?cat?caa?ggc?????825
Ser?Gly?Met?Gln?Asn?Gln?Gln?His?Gln?Phe?Ala?Pro?Gln?His?Gln?Gly
165?????????????????170?????????????????175
caa?cct?caa?gga?cca?ggt?caa?aca?cct?caa?ccg?cca?ggt?tct?gca?act?????873
Gln?Pro?Gln?Gly?Pro?Gly?Gln?Thr?Pro?Gln?Pro?Pro?Gly?Ser?Ala?Thr
180?????????????????185?????????????????190
aac?gct?aat?ttc?cct?atc?aat?atg?cca?cca?aat?ctg?aat?cct?caa?caa?????921
Asn?Ala?Asn?Phe?Pro?Ile?Asn?Met?Pro?Pro?Asn?Leu?Asn?Pro?Gln?Gln
195?????????????????200?????????????????205
caa?atg?ttc?ccc?att?aat?caa?caa?ttt?gct?cag?atg?cca?aat?ggt?caa?????969
Gln?Met?Phe?Pro?Ile?Asn?Gln?Gln?Phe?Ala?Gln?Met?Pro?Asn?Gly?Gln
210?????????????????215?????????????????220
aat?cag?cct?tca?atg?gaa?caa?caa?caa?aga?atg?gca?atg?atg?atg?aaa????1017
Asn?Gln?Pro?Ser?Met?Glu?Gln?Gln?Gln?Arg?Met?Ala?Met?Met?Met?Lys
225?????????????????230?????????????????235?????????????????240
caa?caa?gca?atg?gct?gca?caa?aga?caa?caa?atc?cca?atg?aat?ggt?tta????1065
Gln?Gln?Ala?Met?Ala?Ala?Gln?Arg?Gln?Gln?Ile?Pro?Met?Asn?Gly?Leu
245?????????????????250?????????????????255
gat?cca?caa?caa?caa?cag?caa?atg?atg?aat?gct?gta?ggt?ggt?gga?cct????1113
Asp?Pro?Gln?Gln?Gln?Gln?Gln?Met?Met?Asn?Ala?Val?Gly?Gly?Gly?Pro
260?????????????????265?????????????????270
ggt?aat?ttg?aat?ttg?caa?caa?caa?cta?ttt?tta?caa?caa?caa?cag?cag????1161
Gly?Asn?Leu?Asn?Leu?Gln?Gln?Gln?Leu?Phe?Leu?Gln?Gln?Gln?Gln?Gln
275?????????????????280?????????????????285
cag?cag?cag?caa?ccc?aaa?act?act?ttc?cag?caa?caa?gca?caa?aat?caa????1209
Gln?Gln?Gln?Gln?Pro?Lys?Thr?Thr?Phe?Gln?Gln?Gln?Ala?Gln?Asn?Gln
290?????????????????295?????????????????300
atg?aac?aat?ttg?cgt?caa?caa?gct?gca?atg?gtt?gct?cag?cag?cag?caa????1257
Met?Asn?Asn?Leu?Arg?Gln?Gln?Ala?Ala?Met?Val?Ala?Gln?Gln?Gln?Gln
305?????????????????310?????????????????315?????????????????320
cag?caa?caa?caa?cag?caa?caa?caa?cag?ggt?cag?ttg?caa?ggc?aat?ttg????1305
Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gly?Gln?Leu?Gln?Gly?Asn?Leu
325?????????????????330?????????????????335
gca?tca?gca?atg?ggt?gat?tca?tct?ctg?aag?aat?aac?tct?cct?gtt?ggt????1353
Ala?Ser?Ala?Met?Gly?Asp?Ser?Ser?Leu?Lys?Asn?Asn?Ser?Pro?Val?Gly
340?????????????????345?????????????????350
gca?aga?tca?aat?caa?cag?ctg?act?cca?caa?caa?aat?gct?gca?cca?gct????1401
Ala?Arg?Ser?Asn?Gln?Gln?Leu?Thr?Pro?Gln?Gln?Asn?Ala?Ala?Pro?Ala
355?????????????????360?????????????????365
cca?ctg?cca?cat?cct?tct?caa?caa?ggt?caa?gca?caa?gct?caa?cat?aat????1449
Pro?Leu?Pro?His?Pro?Ser?Gln?Gln?Gly?Gln?Ala?Gln?Ala?Gln?His?Asn
370?????????????????375?????????????????380
ttc?cag?agc?caa?caa?caa?caa?caa?caa?cag?caa?atg?act?aag?atg?gct????1497
Phe?Gln?Ser?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Met?Thr?Lys?Met?Ala
385?????????????????390?????????????????395?????????????????400
ggg?tct?caa?gga?atg?aaa?aag?aat?ggt?cag?atg?tca?aac?ggc?act?agt????1545
Gly?Ser?Gln?Gly?Met?Lys?Lys?Asn?Gly?Gln?Met?Ser?Asn?Gly?Thr?Ser
405?????????????????410?????????????????415
aat?aac?agt?agt?ggc?aga?aac?aat?aat?gct?cta?cga?gat?tat?caa?aat????1593
Asn?Asn?Ser?Ser?Gly?Arg?Asn?Asn?Asn?Ala?Leu?Arg?Asp?Tyr?Gln?Asn
420?????????????????425?????????????????430
caa?tta?atg?tta?tta?gaa?aga?cag?aat?aaa?gag?aga?tta?gaa?ttt?gct????1641
Gln?Leu?Met?Leu?Leu?Glu?Arg?Gln?Asn?Lys?Glu?Arg?Leu?Glu?Phe?Ala
435?????????????????440?????????????????445
aga?aac?act?ggt?aat?tcc?gac?tcc?aac?cca?ttg?agt?aat?gga?atg?atg????1689
Arg?Asn?Thr?Gly?Asn?Ser?Asp?Ser?Asn?Pro?Leu?Ser?Asn?Gly?Met?Met
450?????????????????455?????????????????460
ttt?gcc?ggt?caa?aat?caa?tat?tca?aat?tca?aat?caa?aat?caa?aat?caa????1737
Phe?Ala?Gly?Gln?Asn?Gln?Tyr?Ser?Asn?Ser?Asn?Gln?Asn?Gln?Asn?Gln
465?????????????????470?????????????????475?????????????????480
ctt?cct?cct?aat?caa?caa?caa?cca?act?cca?gca?act?ttc?cac?cca?cca????1785
Leu?Pro?Pro?Asn?Gln?Gln?Gln?Pro?Thr?Pro?Ala?Thr?Phe?His?Pro?Pro
485?????????????????490?????????????????495
cct?ccg?cca?aca?act?gca?aat?ggt?cct?cag?gga?caa?ttt?aat?caa?aaa????1833
Pro?Pro?Pro?Thr?Thr?Ala?Asn?Gly?Pro?Gln?Gly?Gln?Phe?Asn?Gln?Lys
500?????????????????505?????????????????510
cca?tca?cct?gca?acg?tca?aac?aat?tca?cct?gca?tta?ggc?aat?aaa?tca????1881
Pro?Ser?Pro?Ala?Thr?Ser?Asn?Asn?Ser?Pro?Ala?Leu?Gly?Asn?Lys?Ser
515?????????????????520?????????????????525
tca?cca?gca?atg?ggc?aat?aag?aaa?tcg?aag?aaa?gaa?tcc?aat?agt?aaa????1929
Ser?Pro?Ala?Met?Gly?Asn?Lys?Lys?Ser?Lys?Lys?Glu?Ser?Asn?Ser?Lys
530?????????????????535?????????????????540
aag?ggt?aaa?aag?gcg?aat?tca?aat?gct?tct?aca?aca?gca?aac?aac?aaa????1977
Lys?Gly?Lys?Lys?Ala?Asn?Ser?Asn?Ala?Ser?Thr?Thr?Ala?Asn?Asn?Lys
545?????????????????550?????????????????555?????????????????560
aca?tct?gga?caa?aca?aca?cca?aac?atg?tca?caa?cct?cct?agt?gct?ggc????2025
Thr?Ser?Gly?Gln?Thr?Thr?Pro?Asn?Met?Ser?Gln?Pro?Pro?Ser?Ala?Gly
565?????????????????570?????????????????575
acg?gag?cca?aag?cag?cct?caa?cca?aca?gag?caa?atg?cgt?caa?tta?caa????2073
Thr?Glu?Pro?Lys?Gln?Pro?Gln?Pro?Thr?Glu?Gln?Met?Arg?Gln?Leu?Gln
580?????????????????585?????????????????590
gac?aag?caa?cag?cgt?cca?ggt?tca?aat?act?cca?agt?atg?gga?aag?aag????2121
Asp?Lys?Gln?Gln?Arg?Pro?Gly?Ser?Asn?Thr?Pro?Ser?Met?Gly?Lys?Lys
595?????????????????600?????????????????605
gat?ttc?cag?cca?ttg?aca?cct?cgc?tct?gaa?cca?att?agt?ggt?gaa?act????2169
Asp?Phe?Gln?Pro?Leu?Thr?Pro?Arg?Ser?Glu?Pro?Ile?Ser?Gly?Glu?Thr
610?????????????????615?????????????????620
acg?aaa?aag?aag?cgt?aaa?tca?ggt?aaa?ttg?aat?gac?aat?aat?gaa?aat????2217
Thr?Lys?Lys?Lys?Arg?Lys?Ser?Gly?Lys?Leu?Asn?Asp?Asn?Asn?Glu?Asn
625?????????????????630?????????????????635?????????????????640
agt?aat?ggc?aat?tct?cca?aag?aag?caa?gcc?aaa?acc?aat?gca?aac?tcc????2265
Ser?Asn?Gly?Asn?Ser?Pro?Lys?Lys?Gln?Ala?Lys?Thr?Asn?Ala?Asn?Ser
645?????????????????650?????????????????655
aaa?aac?tta?gat?cct?ata?ata?aaa?gaa?gaa?gag?aat?gga?gta?tta?tct????2313
Lys?Asn?Leu?Asp?Pro?Ile?Ile?Lys?Glu?Glu?Glu?Asn?Gly?Val?Leu?Ser
660?????????????????665?????????????????670
ttg?aag?aaa?gaa?tct?tca?act?tca?ttg?caa?gat?caa?gat?cta?gat?tta????2361
Leu?Lys?Lys?Glu?Ser?Ser?Thr?Ser?Leu?Gln?Asp?Gln?Asp?Leu?Asp?Leu
675?????????????????680?????????????????685
aac?ccc?cca?ttg?gca?cca?act?caa?gcc?act?gct?atg?tcc?aat?aca?ttt????2409
Asn?Pro?Pro?Leu?Ala?Pro?Thr?Gln?Ala?Thr?Ala?Met?Ser?Asn?Thr?Phe
690?????????????????695?????????????????700
aac?gac?gat?cca?ttt?gat?gtt?cat?tta?tta?gat?aca?caa?cat?cat?cac????2457
Asn?Asp?Asp?Pro?Phe?Asp?Val?His?Leu?Leu?Asp?Thr?Gln?His?His?His
705?????????????????710?????????????????715?????????????????720
caa?caa?aat?agc?aac?aac?agc?aat?cat?aat?cgt?ggg?caa?aat?ctt?tca????2505
Gln?Gln?Asn?Ser?Asn?Asn?Ser?Asn?His?Asn?Arg?Gly?Gln?Asn?Leu?Ser
725?????????????????730?????????????????735
aat?gga?agt?aat?aat?ctc?agt?gta?agt?ggc?cca?gga?atg?gga?atg?aat????2553
Asn?Gly?Ser?Asn?Asn?Leu?Ser?Val?Ser?Gly?Pro?Gly?Met?Gly?Met?Asn
740?????????????????745?????????????????750
aat?ctg?gta?ttt?ggt?gat?tcg?act?cat?gca?ttt?gac?att?aat?ttc?aac????2601
Asn?Leu?Val?Phe?Gly?Asp?Ser?Thr?His?Ala?Phe?Asp?Ile?Asn?Phe?Asn
755?????????????????760?????????????????765
att?gat?agt?ctt?gat?gat?ata?tgg?aca?act?act?gga?cca?gga?ggt?gat????2649
Ile?Asp?Ser?Leu?Asp?Asp?Ile?Trp?Thr?Thr?Thr?Gly?Pro?Gly?Gly?Asp
770?????????????????775?????????????????780
att?act?ggc?act?ggt?tcg?ggt?tca?gga?ggt?gct?ggc?ggt?acc?gat?gat????2697
Ile?Thr?Gly?Thr?Gly?Ser?Gly?Ser?Gly?Gly?Ala?Gly?Gly?Thr?Asp?Asp
785?????????????????790?????????????????795?????????????????800
gat?aat?ttc?atg?gga?atg?aat?tgg?gct?gca?gat?cca?att?gaa?aat?ggc????2745
Asp?Asn?Phe?Met?Gly?Met?Asn?Trp?Ala?Ala?Asp?Pro?Ile?Glu?Asn?Gly
805?????????????????810?????????????????815
gat?tagaggagtt?tttgaatttt?tttgtattaa?tttttttggt?caagtaaggt?????????2798
Asp
acacttgttt?ggttccaccc?cacccgaccc?gaccccattt?cgccccccac?tggtactggt??2858
ttctatttgt?tccattgtta?tattccttgt?gtttgtttat?ttttcgtttt?ctttattaaa??2918
aattcttttt?ttatttttat?tttcattttc?aattttccat?tttccatttt?tgattttcga??2978
ttttttgttt?tgtatttgta?tttattctta?tcctttattc?cttattcctt?attcttggtc??3038
attttaattt?gaagtttcgg?ttggcttatt?aatttttaat?tgaattatat?tgttaaatta??3098
tattatatta?tattttaatt?tatactatat?atcatttgtt?atattattat?gctattttgt??3158
tttattttaa?atattcaaga?ttttatatct?atatacatac?aattactaat?cattgtttat??3218
agagaaattt?acgaccacat?attgatcctt?ttatcattta?tcaaattgtt?cataatacga??3278
ttgttatgtc?tattgtttgt?aattatttat?aactgatatc?catatatgac?atgtctggtc??3338
cggtattaaa?cttttttatt?cttgtttctt?tttattgttc?ttgttttcat?tagaagtaga??3398
tgataaattg?actgatggtg?ggaacacaat???????????????????????????????????3428
<210>2
<211>817
<212>PRT
＜213〉Candida albicans (Candida albicans)
<400>2
Met?Asn?His?Lys?Gln?Val?Leu?Pro?Glu?Asn?Glu?Pro?Ile?Ser?Thr?Thr
1???????????????5???????????????????10??????????????????15
Thr?Ala?Thr?Pro?Ser?Ser?Ser?Asn?Thr?Met?Val?Pro?Asn?Thr?Thr?Lys
20??????????????????25??????????????????30
Gln?Val?Leu?Asn?Ser?Leu?Ile?Leu?Asp?Phe?Leu?Val?Lys?His?Gln?Phe
35??????????????????40??????????????????45
Gln?Asp?Thr?Ala?Lys?Ala?Phe?Ser?Lys?Glu?Ser?Pro?Asn?Leu?Pro?Ser
50??????????????????55??????????????????60
Ile?Pro?Pro?Leu?Met?Asp?Cys?Ser?Gln?Gly?Phe?Leu?Leu?Glu?Trp?Trp
65??????????????????70??????????????????75??????????????????80
Gln?Val?Phe?Phe?Asp?Leu?Phe?Gln?Val?Arg?Tyr?Gly?Asp?Gly?Asn?Ser
85??????????????????90??????????????????95
Asn?Asn?Asn?Pro?Asn?Asn?Lys?Leu?Tyr?His?Asp?Tyr?Leu?Arg?Val?Gln
100?????????????????105?????????????????110
Glu?Thr?Gln?Lys?His?Leu?Phe?Ser?Gln?Leu?Pro?Leu?Ile?Gln?Gln?Gln
115?????????????????120?????????????????125
Gln?Gln?Gln?Gln?His?His?Phe?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gly?Gln
130?????????????????135?????????????????140
Gln?Gly?Gln?Pro?Phe?Leu?Gln?Gln?Gln?Gln?Arg?Gly?Ile?Gly?Val?Ala
145?????????????????150?????????????????155?????????????????160
Ser?Gly?Met?Gln?Asn?Gln?Gln?His?Gln?Phe?Ala?Pro?Gln?His?Gln?Gly
165?????????????????170?????????????????175
Gln?Pro?Gln?Gly?Pro?Gly?Gln?Thr?Pro?Gln?Pro?Pro?Gly?Ser?Ala?Thr
180?????????????????185?????????????????190
Asn?Ala?Asn?Phe?Pro?Ile?Asn?Met?Pro?Pro?Asn?Leu?Asn?Pro?Gln?Gln
195?????????????????200?????????????????205
Gln?Met?Phe?Pro?Ile?Asn?Gln?Gln?Phe?Ala?Gln?Met?Pro?Asn?Gly?Gln
210?????????????????215?????????????????220
Asn?Gln?Pro?Ser?Met?Glu?Gln?Gln?Gln?Arg?Met?Ala?Met?Met?Met?Lys
225?????????????????230?????????????????235?????????????????240
Gln?Gln?Ala?Met?Ala?Ala?Gln?Arg?Gln?Gln?Ile?Pro?Met?Asn?Gly?Leu
245?????????????????250?????????????????255
Asp?Pro?Gln?Gln?Gln?Gln?Gln?Met?Met?Asn?Ala?Val?Gly?Gly?Gly?Pro
260?????????????????265?????????????????270
Gly?Asn?Leu?Asn?Leu?Gln?Gln?Gln?Leu?Phe?Leu?Gln?Gln?Gln?Gln?Gln
275?????????????????280?????????????????285
Gln?Gln?Gln?Gln?Pro?Lys?Thr?Thr?Phe?Gln?Gln?Gln?Ala?Gln?Asn?Gln
290?????????????????295?????????????????300
Met?Asn?Asn?Leu?Arg?Gln?Gln?Ala?Ala?Met?Val?Ala?Gln?Gln?Gln?Gln
305?????????????????310?????????????????315?????????????????320
Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gly?Gln?Leu?Gln?Gly?Asn?Leu
325?????????????????330?????????????????335
Ala?Ser?Ala?Met?Gly?Asp?Ser?Ser?Leu?Lys?Asn?Asn?Ser?Pro?Val?Gly
340?????????????????345?????????????????350
Ala?Arg?Ser?Asn?Gln?Gln?Leu?Thr?Pro?Gln?Gln?Asn?Ala?Ala?Pro?Ala
355?????????????????360?????????????????365
Pro?Leu?Pro?His?Pro?Ser?Gln?Gln?Gly?Gln?Ala?Gln?Ala?Gln?His?Asn
370?????????????????375?????????????????380
Phe?Gln?Ser?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Gln?Met?Thr?Lys?Met?Ala
385?????????????????390?????????????????395?????????????????400
Gly?Ser?Gln?Gly?Met?Lys?Lys?Asn?Gly?Gln?Met?Ser?Asn?Gly?Thr?Ser
405?????????????????410?????????????????415
Asn?Asn?Ser?Ser?Gly?Arg?Asn?Asn?Asn?Ala?Leu?Arg?Asp?Tyr?Gln?Asn
420?????????????????425?????????????????430
Gln?Leu?Met?Leu?Leu?Glu?Arg?Gln?Asn?Lys?Glu?Arg?Leu?Glu?Phe?Ala
435?????????????????440?????????????????445
Arg?Asn?Thr?Gly?Asn?Ser?Asp?Ser?Asn?Pro?Leu?Ser?Asn?Gly?Met?Met
450?????????????????455?????????????????460
Phe?Ala?Gly?Gln?Asn?Gln?Tyr?Ser?Asn?Ser?Asn?Gln?Asn?Gln?Asn?Gln
465?????????????????470?????????????????475?????????????????480
Leu?Pro?Pro?Asn?Gln?Gln?Gln?Pro?Thr?Pro?Ala?Thr?Phe?His?Pro?Pro
485?????????????????490?????????????????495
Pro?Pro?Pro?Thr?Thr?Ala?Asn?Gly?Pro?Gln?Gly?Gln?Phe?Asn?Gln?Lys
500?????????????????505?????????????????510
Pro?Ser?Pro?Ala?Thr?Ser?Asn?Asn?Ser?Pro?Ala?Leu?Gly?Asn?Lys?Ser
515?????????????????520?????????????????525
Ser?Pro?Ala?Met?Gly?Asn?Lys?Lys?Ser?Lys?Lys?Glu?Ser?Asn?Ser?Lys
530?????????????????535?????????????????540
Lys?Gly?Lys?Lys?Ala?Asn?Ser?Asn?Ala?Ser?Thr?Thr?Ala?Asn?Asn?Lys
545?????????????????550?????????????????555?????????????????560
Thr?Ser?Gly?Gln?Thr?Thr?Pro?Asn?Met?Ser?Gln?Pro?Pro?Ser?Ala?Gly
565?????????????????570?????????????????575
Thr?Glu?Pro?Lys?Gln?Pro?Gln?Pro?Thr?Glu?Gln?Met?Arg?Gln?Leu?Gln
580?????????????????585?????????????????590
Asp?Lys?Gln?Gln?Arg?Pro?Gly?Ser?Asn?Thr?Pro?Ser?Met?Gly?Lys?Lys
595?????????????????600?????????????????605
Asp?Phe?Gln?Pro?Leu?Thr?Pro?Arg?Ser?Glu?Pro?Ile?Ser?Gly?Glu?Thr
610?????????????????615?????????????????620
Thr?Lys?Lys?Lys?Arg?Lys?Ser?Gly?Lys?Leu?Asn?Asp?Asn?Asn?Glu?Asn
625?????????????????630?????????????????635?????????????????640
Ser?Asn?Gly?Asn?Ser?Pro?Lys?Lys?Gln?Ala?Lys?Thr?Asn?Ala?Asn?Ser
645?????????????????650?????????????????655
Lys?Asn?Leu?Asp?Pro?Ile?Ile?Lys?Glu?Glu?Glu?Asn?Gly?Val?Leu?Ser
660?????????????????665?????????????????670
Leu?Lys?Lys?Glu?Ser?Ser?Thr?Ser?Leu?Gln?Asp?Gln?Asp?Leu?Asp?Leu
675?????????????????680?????????????????685
Asn?Pro?Pro?Leu?Ala?Pro?Thr?Gln?Ala?Thr?Ala?Met?Ser?Asn?Thr?Phe
690?????????????????695?????????????????700
Asn?Asp?Asp?Pro?Phe?Asp?Val?His?Leu?Leu?Asp?Thr?Gln?His?His?His
705?????????????????710?????????????????715?????????????????720
Gln?Gln?Asn?Ser?Asn?Asn?Ser?Asn?His?Asn?Arg?Gly?Gln?Asn?Leu?Ser
725?????????????????730?????????????????735
Asn?Gly?Ser?Asn?Asn?Leu?Ser?Val?Ser?Gly?Pro?Gly?Met?Gly?Met?Asn
740?????????????????745?????????????????750
Asn?Leu?Val?Phe?Gly?Asp?Ser?Thr?His?Ala?Phe?Asp?Ile?Asn?Phe?Asn
755?????????????????760?????????????????765
Ile?Asp?Ser?Leu?Asp?Asp?Ile?Trp?Thr?Thr?Thr?Gly?Pro?Gly?Gly?Asp
770?????????????????775?????????????????780
Ile?Thr?Gly?Thr?Gly?Ser?Gly?Ser?Gly?Gly?Ala?Gly?Gly?Thr?Asp?Asp
785?????????????????790?????????????????795?????????????????800
Asp?Asn?Phe?Met?Gly?Met?Asn?Trp?Ala?Ala?Asp?Pro?Ile?Glu?Asn?Gly
805?????????????????810?????????????????815
Asp
<210>3
<211>30
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(30)
＜223〉primer
<400>3
ctgggattca?tgaatcataa?acaagtacta??????????????????????????????????????30
<210>4
<211>31
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(31)
＜223〉primer
<400>4
ctgcgcatgc?atcgccattt?tcaattggat?c????????????????????????????????????31
Claims (8)
1. isolating Candida albicans CaFL08 polypeptide is characterized in that this polypeptide is selected from down group:
(a) has the polypeptide of SEQ ID NO:2 aminoacid sequence;
(b) SEQ ID NO:2 aminoacid sequence is formed through replacement, disappearance or the interpolation of one or more amino-acid residues, and have transcriptional activation function by (a) polypeptides derived.
2. polypeptide as claimed in claim 1 is characterized in that, it has the aminoacid sequence shown in the SEQ ID NO:2.
3. isolating polynucleotide is characterized in that, it comprises a nucleotide sequence, and this nucleotide sequence is selected from down group:
(a) polynucleotide of polypeptide as claimed in claim 1 or 2 of encoding;
(b) with polynucleotide (a) complementary polynucleotide.
4. polynucleotide as claimed in claim 3 is characterized in that this polynucleotide encoding has the polypeptide of aminoacid sequence shown in the SEQ IDNO:2.
5. polynucleotide as claimed in claim 4 is characterized in that, the sequence of these polynucleotide has 298-2748 position among the SEQID NO:1.
6. a carrier is characterized in that, it contains the described polynucleotide of claim 3.
7. a host cell is characterized in that, it contains the described carrier of claim 6.
8. energy and the described Candida albicans CaFL08 of claim 1 protein-specific bonded antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB03150552XA CN100519578C (en) | 2003-08-25 | 2003-08-25 | White candidas transcription factor gene and its use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB03150552XA CN100519578C (en) | 2003-08-25 | 2003-08-25 | White candidas transcription factor gene and its use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1590405A true CN1590405A (en) | 2005-03-09 |
| CN100519578C CN100519578C (en) | 2009-07-29 |
Family
ID=34597582
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB03150552XA Expired - Fee Related CN100519578C (en) | 2003-08-25 | 2003-08-25 | White candidas transcription factor gene and its use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100519578C (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101712935B (en) * | 2008-10-08 | 2013-04-10 | 中国科学院上海生命科学研究院 | Candida albicans toxicity related gene CaMSS11 and application thereof |
| CN110229221A (en) * | 2019-06-27 | 2019-09-13 | 上海交通大学医学院附属仁济医院 | It is a kind of for detecting the antigen and application thereof of aggressive candidiasis |
-
2003
- 2003-08-25 CN CNB03150552XA patent/CN100519578C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101712935B (en) * | 2008-10-08 | 2013-04-10 | 中国科学院上海生命科学研究院 | Candida albicans toxicity related gene CaMSS11 and application thereof |
| CN110229221A (en) * | 2019-06-27 | 2019-09-13 | 上海交通大学医学院附属仁济医院 | It is a kind of for detecting the antigen and application thereof of aggressive candidiasis |
| CN110229221B (en) * | 2019-06-27 | 2022-04-12 | 上海交通大学医学院附属仁济医院 | Antigen for detecting invasive candidiasis and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100519578C (en) | 2009-07-29 |
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