KR102518785B1 - Phosphatase that modulates cell membrane stability of Cryptococcus neoformans, and method for screening antifungal agents using the same - Google Patents

Abstract

VPS29, YMR1, SSU72, NEM1, YVH1, SIT4, GUA1, INP5201, PHS1, DBR1, SIW14, SDP102, OCA1, GDA1, FBP26, PSR1, CDC1, INP5202, and YND1 are associated with the pathogenicity of Cryptococcus neoformans and phosphatase genes whose functions of regulating cell membrane stability have been newly identified, and can be used for screening new antifungal agents targeting these genes.

Description

Phosphatase that modulates cell membrane stability of Cryptococcus neoformans, and method for screening antifungal agents using the same}

The present invention relates to a phosphatase that regulates cell membrane stability of Cryptococcus neoformans and a method for screening antifungal agents using the same.

All living organisms dynamically respond to changing environments, and these responses are achieved by complex signal transduction pathways. When an environmental cue occurs, the organism receives the signal through sensors such as receptor proteins and activates downstream effectors to respond to the ensuing stress. When an environmental signal disappears or an organism adapts to the environment, the corresponding signaling pathway is usually desensitized in time and can be reactivated later. The process of activation and inactivation of these signal transduction pathways is in many cases achieved by phosphorylation by kinase and dephosphorylation by phosphatase, which turns signal transduction on and off. It is a post-translational modification. Therefore, regulation of kinases and phosphatases in signal transduction pathways is important to maintain cellular homeostasis.

Pathogenic fungi utilize these signaling pathways to adapt to environmental changes that occur during infection, colonization, proliferation, and dissemination in various tissues within the host.

Cryptococcus neoformans causes meningitis in immunocompromised patients and causes more than 180,000 deaths worldwide each year. A large-scale functional analysis of Cryptococcus neoformans transcription factors (TFs) and kinases identified 178 TFs and 183 kinases. Of these, 155 TFs and 129 kinases were functionally characterized in vitro and in vivo, and 45 TFs and 63 kinases were known to be involved in pathogenicity. Nevertheless, it has not been clearly clarified how the signaling components are regulated and coordinated to exhibit pathogenicity.

In order to clearly identify TF and kinase networks, joint analysis of their corresponding phosphatase networks is essential. In the past, phosphatases have not received attention because kinases with high substrate specificity have been considered more suitable as drug targets.

However, recent functional phosphatome data revealed in several types of pathogenic fungi revealed that phosphatase plays an important role in maintaining homeostasis by regulating growth, cell cycle, differentiation, stress response and metabolism of fungi. lost.

In ascomycete, 32 phosphatases of Aspergillus fumigatus were identified by genome-wide analyses, and 24 of them were functionally characterized. , it has been identified that most of them contribute to stress response, iron assimilation, toxin production, and resistance.

In addition, 82 phosphatase genes were identified in Fusarium graminearum, which causes wheat scab, and 71 of them were destroyed and functionally analyzed through phenotypic characteristics of 15 species. It was confirmed that phosphatase is involved in the virulence of plant pathogenic fungi. (Yun, Y. et al. Functional analysis of the Fusarium graminearum phosphatome. New Phytol. 207, 119-134 (2015).)

However, in Cryptococcus neoformans, nothing is known at the genomic level except for some few genes in phosphatase associated with pathogenicity.

(Non-Patent Document 0001) Yun, Y. et al. Functional analysis of the Fusarium graminearum phosphatome. New Phytol. 207, 119-134 (2015).

According to one embodiment, a phosphatase gene whose relevance to pathogenicity has been newly identified as a phosphatase that regulates cell membrane stability of Cryptococcus neoformans and an antifungal agent screening method using the same are provided.

In one aspect , VPS29, YMR1, SSU72, NEM1, YVH1, SIT4, GUA1, INP5201, PHS1, DBR1, SIW14, SDP102, OCA1, GDA1, contacting mRNA of at least one gene selected from the group consisting of FBP26, PSR1, CDC1, INP5202, and YND1 or a protein expressed therefrom with a candidate material; Measuring the expression level of the mRNA or protein, or measuring the activity of the protein; and determining that the candidate material has antifungal activity when the measured expression level or activity is significantly reduced compared to the control group.

The present inventors selected 31 genes whose virulence to Galleria mellonella larvae was reduced or infectivity to murine was reduced when inactivated among 139 candidate phosphatase genes of the pathogenic fungus Cryptococcus neoformans. Among them, 21 showed changes in 7 types of phenotypes related to pathogenicity ( O -sugar chain formation, cell membrane stability, DNA damage response, blood-brain barrier passage, capsule formation, melanin biosynthesis, and thermolerance for mammalian body temperatures). Phosphatases were selected. The final selected gene is a phosphatase gene that has been newly identified as regulating expression traits closely related to the virulence and infectivity of Cryptococcus neoformans, and can be a new target for antifungal agents. The 21 phosphatase genes finally selected are disclosed in Table 1 below.

SEQ ID H99 lD Name One 00182 VPS29 2 00390 YMR1 3 01054 SSU72 4 01177 NEM1 5 01203 YVH1 6 01254 OCA101 7 01436 SIT4 8 01877 GUA1 9 02109 INP5201 10 02487 PHS1 11 02490 MRE11 12 03222 DBR1 13 03296 SIW14 14 03893 SDP102 15 03963 OCA1 16 03996 GDA1 17 04221 FBP26 18 04224 PSR1 19 06647 CDC1 20 07317 INP5202 21 07692 YND1

The Cryptococcus fungus is, for example, a pathogenic fungus Cryptococcus neoformans , Cryptococcus gattii , Cryptococcus albidus , or Cryptococcus uniguttulatus ) can be.

The antifungal activity may be against a fungus belonging to the genus Cryptococcus or a pathogenic fungus belonging to the genus Cryptococcus.

The VPS29 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 1 or a gene composed of the nucleotide sequence of SEQ ID NO: 22.

The YMR1 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 2 or a gene having the nucleotide sequence of SEQ ID NO: 23.

The SSU72 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 3 or a gene composed of the nucleotide sequence of SEQ ID NO: 24.

The NEM1 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 4 or a gene composed of the nucleotide sequence of SEQ ID NO: 25.

The YVH1 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 5 or a gene composed of the nucleotide sequence of SEQ ID NO: 26.

The SIT4 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 7 or a gene composed of the nucleotide sequence of SEQ ID NO: 28.

The GUA1 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 8 or a gene composed of the nucleotide sequence of SEQ ID NO: 29.

The INP5201 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 9 or a gene having the nucleotide sequence of SEQ ID NO: 30.

The PHS1 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 10 or a gene composed of the nucleotide sequence of SEQ ID NO: 31.

The DBR1 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 12 or a gene having the nucleotide sequence of SEQ ID NO: 33.

The SIW14 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 13 or a gene having the nucleotide sequence of SEQ ID NO: 34.

The SDP102 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 14 or a gene having the nucleotide sequence of SEQ ID NO: 35.

The OCA1 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 15 or a gene composed of the nucleotide sequence of SEQ ID NO: 36.

The GDA1 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 16 or a gene having the nucleotide sequence of SEQ ID NO: 37.

The FBP26 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 17 or a gene composed of the nucleotide sequence of SEQ ID NO: 38.

The PSR1 gene may be a gene encoding a phosphatase composed of the amino acid sequence of SEQ ID NO: 18 or a gene composed of the nucleotide sequence of SEQ ID NO: 39.

The CDC1 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 19 or a gene having the nucleotide sequence of SEQ ID NO: 40.

The INP5202 gene may be a gene encoding a phosphatase having the amino acid sequence of SEQ ID NO: 20 or a gene having the nucleotide sequence of SEQ ID NO: 41.

The expression reduction candidate material may include, but is not limited to, a small molecule compound, antibody, antisense nucleotide, siRNA, dsRNA, nucleic acid, protein, peptide, extract, or natural product.

The contacting step may include contacting the isolated mRNA or protein with the candidate substance, or contacting a Cryptococcus genus fungus expressing the gene with the candidate substance. In the case of contacting a fungus of the genus Cryptococcus with a candidate substance, it may be performed by culturing for a time sufficient for the candidate substance to penetrate into the fungus. The contacting step may be performed in vitro.

Measurement of the mRNA expression level can be performed by a method well known to those skilled in the art, for example, reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction, real-time reverse transcription polymerase reaction, RNase protection assay (RPA) , Northern blotting, DNA chip (Microarray) or RNA sequencing analysis.

The expression level of the protein can be measured by a method well known to those skilled in the art, for example, Western blot, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, tissue Immunoprecipitation Assay, Complement Fixation Assay, Fluorescence Activated Cell Sorter (FACS), or protein chip.

The candidate substance for inhibiting activity may include, but is not limited to, a small molecule compound, antibody, antisense nucleotide, siRNA, dsRNA, nucleic acid, protein, peptide, extract, or natural product.

To measure the activity of the protein, an enzyme activity measurement method known to those skilled in the art may be used, for example, measuring the degree of phosphorylation of a phosphatase substrate using labeled phosphoric acid, or using an antibody specific for the phosphorylation site of a substrate. A method for quantitatively analyzing the degree of binding can be used.

The screening method according to one embodiment uses a phosphatase gene whose relevance to the pathogenicity of Cryptococcus neoformans is newly identified, so that a new antifungal agent can be screened.

Figure 1 shows in vitro phenotypic changes and in vivo expression profiling for each cluster in vitro by phosphatase deletion of C. neoformans. The abbreviations used in FIG. 1 are as follows. 25: 25° C.; 30: 30° C.; 37: 37° C.; 39: 39° C.; CAP: capsule production; MEL: melanin production; URE: urease production; MAT: mating; HPX: hydrogen peroxide; TBH: tert-butyl hydroperoxide; MD: menadione; DIA: diamide; MMS: methyl methanesulphonate; HU: hydroxyurea; 5FC: 5-flucytosine; AMB: amphotericin B; FCZ: fluconazole; FDX: fludioxonil; TM: tunicamycin; DTT: dithiothreitol; CDS: cadmium sulfate; SDS: sodium dodecyl sulfate; CR: Congo red; CFW: calcofluor white; KCR: YPD + 1.5M KCl; NCR: YPD + 1.5 M NaCl; SBR: YPD + 2 M sorbitol; KCS: YP + 1 M KCl; NCS: YP + 1 M NaCl; SBS: YP + 2M sorbitol.
Figure 2 shows the results of the identification of virulence-regulated phosphatase by Galleria mellonella insect lethality assay. (n ≥ 15)
Figure 3 shows the results of identification of infectivity-regulated phosphatase by the STM-based murine infection model.
Figure 4 shows the growth rate at 30 ℃ and 37 ℃ of phosphatase variants.
Figure 5 shows the results of measuring the melanin production of the mutant using three different melanin induction media (Niger seed, dopamine, epinephrine media).
6 is a result of confirming the gene expression level of LAC1, a melanin expression regulating gene, under nutritional conditions (R) and deficient conditions (S).
7 is a result of confirming the gene expression levels of BZP4 and HOB1, which are genes related to melanin expression, under nutrient deficiency conditions (R) and deficiency conditions (S).
8 is a result of capsule production analysis using a capsule induction medium (FBS agar medium). Capsule thickness (total diameter - cell body diameter) was measured in WT cells (n = 50) and each phosphatase variant (n = 50).
9 shows a phenotypic heatmap of retromeric complex-related gene mutations. Deletion genes and strains are vps29 Δ (YSB4881, YSB4882), vps26 Δ (YSB5671, YSB5672), vps35 Δ (YSB5615, YSB5616), vps5 Δ (YSB5683, YSB5684), and vps17 Δ (YSB5724).
10 is a result of insect lethality analysis of retromer complex mutations.
11 shows the results of an STM-based murine infection assay of retromeric complex mutations.
12 shows HPLC profiles of O -linked glycans bound to cell wall mannoproteins of C. neoformans wild type (WT), gda1 Δ (YSB4750), and ynd1 Δ (YSB4856).
Figure 13 shows the O- glycan components of C. neoformans wild type (WT), gda1 Δ (YSB4750), and ynd1 Δ (YSB4856).
14 is a schematic diagram of the functions of Gda1p and Ynd1p predicted from the experimental results.
Figure 15 shows the BBB migration ability of C. neoformans wild type and mutants in an in vitro Blood-Brain Barrier-passing assay (BBB-crossing assay).
16 shows the results of a BBB-adhesion assay using a human brain microvascular endothelial cell line (hCMEC/D3 cell line).
17 shows the expression levels of brain infection-related genes whose expression was induced under host-mimic conditions (HMC) in wild-type and phosphatase mutants of C. neoformans.
18 is a result of generating a functional protein association network by STRING analysis in order to comprehensively understand the signal network related to brain infection by C. neoformans.
19 is a result of analyzing pathogenicity-related phosphatase of pathogenic fungi.
20 and 21 show VPS29, YMR1, SSU72, NEM1, YVH1, SIT4, GUA1, INP5201, PHS1, DBR1, SIW14, SDP102, OCA1, GDA1, FBP26, PSR1, CDC1, INP5202, or YND1 gene deleted C. This is a result confirming that neoformans growth is inhibited in SDS-added medium.

Hereinafter, one or more specific examples will be described in more detail through examples. However, these examples are intended to illustrate one or more specific examples, and the scope of the present invention is not limited to these examples.

The present inventors target 109 of 139 genes presumed to be phosphatase in Cryptococcus neoformans ( Cryptococcus neoformans ), and 219 types of signature-tagged gene-deletion mutant strains ) library was built. Here, 11 signature tag mutants for 6 phosphatases constructed in a previous study (Jung, KW et al. Systematic functional profiling of transcription factor networks in Cryptococcus neoformans . Nat. Commun. 6, 6757 (2015).) used Using 230 types of signature tag mutants constructed for a total of 114 phosphatases, their phenotypes were analyzed in 30 types of in vitro conditions, and toxicity analysis was performed using two experimental models (insect and murine). performed Phosphatase Phenome Data sets are published in the Phosphatase Phenome Database ( http://phosphatase.cryptococcus.org ).

Experiment method

1. Compliance with research ethics regulations (Ethnic statement)

Animal care and all experiments were performed in accordance with the ethical guidelines of the Institutional Animal Care and Use Committee (IACUC) of Yonsei University.

2. Construction of C. neoformans phosphatase and retromer mutants

C. neoformans through homologous recombination using a NAT-split marker and double joint PCR (DJ-PCR), and a gene disruption cassette containing a norseothricin acetyl transferase (NAT) marker Phosphatase and retromer mutant strains were constructed from the S. serotype A H99S strain.

Primers used to construct phosphatase and retromer mutant strains, for example, L1 Primer (5' flanking region primer 1), L2 Primer (5' flanking region primer 2), R1 Primer (3' flanking region primer 1) for each gene For information on region primer 1), R2 Primer (3' flanking region primer 2), SO primer (diagnostic screening primer, pairing with B79), PO primer (Southern blot probe primer), STM primer, and STM common primer, SEQ ID NO: 43 to SEQ ID NO: 211.

Other primers, B1026 (M13 Forward extended), B1027 (M13 Reverse extended), B79 (Screening primer), B1454 (NAT split marker primer 1), B1455 (NAT split marker primer 2), B1886 (NEO split marker primer 1) ), B1887 (NEO split marker primer 2), B679 (qRT-PCR primer for ACT1), B680 qRT-PCR primer for ACT1), B8953 (qRT-PCR primer for LAC1), B8954 (qRT-PCR primer for LAC1), B3737 (qRT-PCR primer for BZP4), B8521 (qRT-PCR primer for BZP4), B6394 (qRT-PCR primer for HOB1), B8519 (qRT-PCR primer for HOB1), B8654 (qRT-PCR primer for ITR1A), B8655 (qRT-PCR primer for ITR1A), B8658 (qRT-PCR primer for ITR3C), B8659 (qRT-PCR primer for ITR3C), B8656 (qRT-PCR primer for MPR1), B8657 (qRT-PCR primer for MPR1), B8598 (qRT-PCR primer for FZC31), B4951 (qRT-PCR primer for FZC31), B9243 (qRT-PCR primer for GAT201), B9422 (qRT-PCR primer for GAT201), B9061 (qRT-PCR primer for PDR802), B4913 (qRT-PCR primer for PDR802) is disclosed in SEQ ID NO: 212 to SEQ ID NO: 238.

Detailed information on the primers and probes is in supplementary data 4 of the present inventor's paper Genome-wide functional analysis of phosphatases in the pathogenic fungus Cryptococcus neoformans , Bahn YS et al., Nature Communications volume 11, Article number: 4212 (2020) can refer to. (https://www.nature.com/articles/s41467-020-18028-0#Sec28)

From H99S genomic DNA, PCR was performed with primer pairs L1/L2 (5' flanking region primer set) and R1/R2 (3' flanking region primer set) to amplify the 5' flanking region and 3' flanking region of the target gene.

The signature tagged NAT markers were amplified by PCR using primers M13Fe (M13 forward extended) and M13Re (M13 reverse extended) from the pNAT-STM plasmid containing the NAT gene with each unique signature tag sequence.

In the first PCR, template DNA was generated by amplifying the 5' flanking region, the 3' flanking region, and the NAT marker, and in the second PCR, primer pairs L1/NSL (B1455 NAT split marker primer 2) and R2/NSR (B1454 The 5'-region and 3'-region of the NAT-split gene-disruption cassette were constructed with NAT split marker primer 1).

Biological transformation introducing the NAT gene disruption cassette was performed in the following manner. The H99S strain was cultured in 50ml YPD medium at 30° C. for 16 hours, followed by spin-down. It was resuspended in 5 ml of distilled water, applied to YPD agar medium containing 1M sorbitol, and further incubated at 30°C for 3 hours. The gene disruption cassette was coupled with 600 μg of 0.6 μm gold microcarrier beads (Bio - Rad Laboratories, Hercules, CA, USA) and introduced into cells using a particle delivery system (PDS-100, Bio - Rad). After culturing at 30° C. for 4 hours to restore cell membrane integrity, the cells were scraped and spread on YPD agar medium containing norseotricin (100 μg/ml). NAT-positive transformants were confirmed by diagnostic PCR. The genotype of each screened transformant was confirmed by Southern blot analysis. At least two independent mutant strains were constructed for each phosphatase gene.

3. nCounter in vivo gene expression profiling analysis.

By performing nCounter gene expression analysis (NanoString) using RNA samples reported in previous studies (Lee, KT et al. Fungal kinases and transcription factors regulating brain infection in Cryptococcus neoformans . Nat. Commun. 11, 1521 (2020)) The in vivo expression levels of 139 phosphatases were quantified.

Total RNA samples were obtained from 6-week-old female A/J mice infected with 5 × 10 ⁵ cells via nasal aspiration. Groups of three mice were each sacrificed at 3, 7, 14, and 21 dpi (day post-infection), and lungs, brains, spleens, and kidneys were obtained and freeze-dried. The dried organs were homogenized and total RNA was extracted with a commercial RNA extraction kit (easy-BLUE, Intron Biotechnology).

A sample containing 10 ng of total RNA isolated from C. neoformans grown in in vitro basal conditions (30 ° C; YPD medium) or 10 μg of total RNA isolated from C. neoformans infected mouse tissues The nCounter multiplex platform (NanoString, Seattle, WA, USA) was set up according to the manufacturer's standard protocol and reacted with a custom probe code set. High resolution (600 fields) was scanned with an optional digital analyzer and normalized with nSolver software (NanoString).

Eight house-keeping genes (mitochondrial protein, CNAG_00279; microtubule-binding protein, CNAG_00816; aldose reductase, CNAG_02722; cofilin, CNAG_02991; actin, CNAG_00483; tubulin β chain, CNAG_01840; tubulin α-1A chain, CNAG_03787; histone H3, CNAG_04828) was used for expression normalization.

Normalized data were transformed into log ₂ scores to represent fold-change and averaged concatenation by Morpheus (Broad Institute, Cambridge, MA, USA, http://software.broadinstitute.org/morpheus ). clustering was performed using average linkage) and one minus Pearson correlation.

4. Growth and Chemosusceptibility Testing

To analyze the susceptibility of each phosphatase mutant to stress conditions in vitro, C. neoformans was grown at 30° C. for 16 h, serially diluted 10-fold (1-10 ⁴ ), and subjected to the following environmental stressors. Spot inoculation was performed on YPD agar medium containing inducing chemicals. (Jung, KW et al. Systematic functional profiling of transcription factor networks in Cryptococcus neoformans . Nat. Commun. 6, 6757 (2015) method reference)

[Environmental Stress Inducing Chemicals]

osmotic stress (sorbitol) and cation/salt stress (NaCl and KCl) under glucose excess (YPD) or glucose deficiency (yeast extract-peptone; YP) conditions;

Oxidative stress [hydrogen peroxide (H ₂ O ₂ ), tert-butyl hydroperoxide (organic peroxide), menadione (superoxide anion generator), diamide (thiolspecific oxidant)];

toxic heavy metal stress [cadmium sulfate (CdSO4)]; genotoxic stress (methyl methanesulphonate and hydroxyurea);

membrane destabilizing stress [sodium dodecyl sulfate (SDS)];

cell wall destabilizing stress (calcofluor white and Congo red);

ER stress [tunicamycin and dithiothreitol (DTT)];

and antifungal drug susceptibility (fludioxonil, fluconazole, amphotericin B, and flucytosine).

Cells were cultured at 30° C. for 1 to 5 days and photographed daily. To investigate the growth of C. neoformans strains at various temperatures, cells serially diluted in YPD agar medium were spot inoculated and cultured at 25°C, 37°C and 39°C, and culture pictures were taken daily. To quantitatively investigate the growth rate of the phosphatase mutants, the WT strain (H99S) and the phosphatase mutants were cultured overnight at 30 °C and subcultured in fresh liquid YPD medium [optical density OD ₆₀₀ = 0.2]. Cells were cultured at 30 °C or 37 °C in a multi-channel bioreactor (Biosan Laboratories, Inc., Warren, MI, USA), and OD 600 nm was automatically measured for 40 to 90 hours.

5. Mating assay

To measure the unilateral mating efficiency, each serotype A MAT α phosphatase mutant constructed with the H99S strain and the MAT a KN99a strain was incubated in YPD medium at 30 ° C. for 16 hours, twice with PBS. Washed. The cells were mixed at the same concentration (10 ⁷ cells/ml), spot-inoculated in V8 mating medium (pH 5), and cultured at 25°C in the dark for 7 to 14 days. Filamentous growth was observed and photographed weekly.

6. In vitro virulence factor production assay

To test capsule production efficiency, each mutant was cultured at 30 °C, spot inoculated on Dulbecco's modified Eagle agar medium and incubated at 37 °C for 2 days. The cultured cells were scrapped, washed with distilled water, fixed with 10% formalin and washed again with distilled water. The fixed cells were adjusted to a concentration of 3×10 ⁸ cells/ml, and 50 μl of the cell suspension was injected into a microhaematocrit capillary tube (Kimble Chase, Rockwood, TN, USA). Capillary tubes were placed vertically for 10 days to pack the cells by gravity. The volume ratio of packed cells (packed cell phase/total phase) was measured, and each ratio was normalized with the packed cell volume of the wild-type H99S strain to determine the relative packed cell volume (packed cell phase) of each mutant. cell volume) was calculated. Statistical differences in relative enriched cell volume proportions were determined by one-way analysis of variance (ANOVA) using Bonferroni's multiple comparison test in Prism 8 (GraphPad, San Diego, USA).

To measure the efficiency of capsule production in different media, 5 μl of each culture medium was spot inoculated into Littman's agar medium and FBS agar medium (10% fetal bovine serum and 90% PBS), cultured at 37°C for 2 days, and scraped cells. and resuspended in distilled water. The resuspended cells were stained with India ink (BactiDrop; Remel, San Diego, CA, USA) and observed under a DIC (Differential Interference Contrast) microscope (BX51, Olympus, Tokyo, Japan). Capsule thickness was determined by subtracting the cell diameter from the capsule diameter (total diameter minus cell body diameter). For quantitative measurement of capsule thickness, 50 cells of the H99S strain and each phosphatase mutant were measured.

To investigate melanin production efficiency, each phosphatase mutant was incubated in YPD medium at 30 °C for 16 hours, washed with PBS, and Niger seed, dopamine, or epinephrine medium containing 0.1% or 0.2% glucose (1g L per liter). - 3 μl of asparagine, 3 g KH ₂ PO ₄ , 250 mg MgSO ₄ , 1 mg thiamine, 5 μg biotin, 100 mg L-DOPA or epinephrine hydrochloride) was spot inoculated. Spot-inoculated cells were cultured at 37° C. and photographs were taken after 1 to 3 days. Phosphatase mutants with growth defects at 37 °C were examined for melanin and capsule production efficiency at 30 °C.

To confirm urease production, each phosphatase mutant was incubated at 30 ° C for 16 hours, washed with PBS, and then inoculated into liquid Christensen's media contained in a 10ml medical tube (SPL Life Sciences, Gyeonggi-do, Korea) (cells 10 ⁶ ), then incubated in a shaking incubator at 30°C for 1 to 3 days, and pictures were taken every day.

7. Expression analysis

To measure the expression levels of melanin regulatory genes ( LAC1, HOB1, MBS1, BZP4 ), the H99S strain and phosphatase mutants ( mre11 Δ, ccr4 Δ, vps29 Δ, yvh1 Δ, fbp26 Δ, inp5201 Δ, cac1 Δ, ptp2 Δ, ptc2 Δ, dbr1 Δ, ppg1 Δ, nem1 Δ and gua1 Δ) were cultured in YPD liquid medium at 30° C. for 16 hours and subcultured into fresh liquid YPD medium (OD ₆₀₀ = 0.2).

A baseline sample was prepared by sampling half of the cell culture when the cells reached the initial logarithmic phase (OD ₆₀₀ = 0.6 to 0.8). The remaining cell cultures were washed three times with PBS and incubated in nutrient starvation conditions (YNB medium containing ammonium sulfate without glucose) for 2 hours, then immediately pelleted in liquid nitrogen and lyophilized.

To measure the expression levels of BBB crossing-related genes ( ITR1A, ITR3C, MPR1, FZC31, GAT201 and PDR802 ), strain H99S and phosphatase mutants [ sit4 Δ (YSB4094), siw14 Δ (YSB4570) , ssu72 Δ (YSB4242), gda1 Δ (YSB4750) and xpp1 Δ (YSB5941)] were cultured in YPD broth at 30° C. for 16 h, subcultured in 50 ml fresh YPD broth, and OD ₆₀₀ reached 0.8. It was further cultured until Then, the culture medium was separated into two 25 ml tubes, centrifuged, and washed three times with sterile distilled water.

One tube was kept in liquid nitrogen to monitor basal expression levels, and the other tube was resuspended in an equal volume of RPMI1640 medium containing 10% FBS. After incubation at 37°C for 3 hours in a CO ₂ incubator with horizontal shaking at 120 rpm, the cells were centrifuged and freeze-dried overnight. Total RNA was extracted from each sample using a commercially available RNA extraction kit (easy-BLUE, iNtRON Biotechnology, Gyeonggi, Korea), and cDNA was synthesized using RTase (Thermo Scientific, Waltham, MA, USA). Quantitative reverse transcription-PCR (qRT-PCR) was performed using target gene-specific primer pairs of SEQ ID NOs: 219 to 238. For the target gene-specific primer pair, reference may be made to Genome-wide functional analysis of phosphatases in the pathogenic fungus Cryptococcus neoformans , Bahn YS et al., Nature Communications volume 11, Article number: 4212 (2020).

8. HPLC analysis of cwMPs (cell wall mannoproteins) O- linked glycans

Structural analysis of N-/O- glycans assembled on proteins in yeasts. J. Microbiol. 56, 11-23 (2018).

O- linked oligosaccharides were released from purified cwMP by modified hydrazinolysis. Dried cwMP (50 μg) was resuspended in hydrazine monohydrate and incubated at 60° C. for 4 hours. The reaction was cooled and desiccation, the pellet was dissolved in NaHCO ₃ , mixed with (CH ₃ CO) ₂ O and incubated on ice for 30 minutes.

O- glycans were purified with Dowex 50WX8-400 resins (Sigma-Aldrich, St. Louis, MO, USA) and labeled with 2-AA. Purified O- glycans were purified using TSKgel Amide-80 using 90% solvent A (2% acetic acid and 1% tetrahydrofuran in acetonitrile) and 10% solvent B (5% acetic acid, 3% triethylamine and 1% tetrahydrofuran in water). Analysis was performed using HPLC on a column (0.46 × 25 cm, Tosoh Corp., Tokyo, Japan). After sample injection, the proportion of solvent B was increased to 90% over 60 minutes at a flow rate of 1.0 ml/min, and O- glycans were detected with a fluorescence detector (2475, Waters Corp., Milford, MA, USA). The excitation and emission wavelengths are 360 nm and 425 nm, respectively. Data were analyzed using chromatography software (Empower 2, Waters).

9. In vivo virulence analysis based on insect model

A minimum of 15 G. mellonella larvae (Vanderhorst Wholesale, Inc., Saint Marys, OH, USA) reaching a body weight of 200-300 mg and in the final larval instar were used. Each phosphatase variant and wild-type H99S strain was incubated overnight at 30° C., pelleted, washed three times with PBS, and resuspended in PBS at a concentration of 10 ⁶ cells/ml. For each larva, 4000 C. neoformans cells were harvested from the second to last prolegs using a 100 μl syringe (PB600-1, Hamilton Company, Reno, NV, USA) equipped with a 10 μl needle and repeat dispenser. ) was injected between Negative control G. mellonella was injected with only PBS. Infected larvae were placed in Petri dishes and monitored daily while incubated at 37°C in a humidified environment. The larvae were judged to be dead when they reached a state in which they turned black and showed no movement when touched. Larvae that pupate during the experiment were screened for statistical analysis. Survival curves were delineated using Prism 8 (GraphPad, San Diego, CA, USA) and analyzed with the log-rank test (Mantel-Cox). Two independent strains were investigated for each phosphatase mutant.

10. STM-based murine infectivity assay

A set of phosphatase mutants with 41 unique signature-tagged NAT selection markers were incubated at 30°C for 16 hours. The ste50 Δ (STM # 282) and ire1 Δ (STM # 169) mutants were used as virulent and avirulent control strains, respectively.

The mutant and control strains were pelleted, washed three times with PBS, resuspended in PBS, and then pooled in equal amounts (5 x 10 ⁵ cells).

7-week-old female A/J mice (Jackson Laboratory, Bar Harbor, ME, USA) anesthetized with intraperitoneal injection of Avertin (2,2,2-tribromoethanol, T48402, Sigma-Aldrich, St. Louis, MO, USA) were infected with pooled mutants (50 μl PBS) of 5×10 ⁵ cells via intranasal inhalation.

To prepare the input phosphatase genomic DNA library, 200 μl of the pooled strains were spread on YPD medium containing 100 μg/ml chloramphenicol and incubated at 30 ° C for 3 days. Collected by scraping.

Infected mice were sacrificed by overdose of anesthetic (Avertin) for humane experiment termination when they reached 14 dpi. Lungs and brains of infected mice were harvested and homogenized with 5 ml of PBS. Then, the homogenized tissues were spread on YPD medium containing 100 μg/ml chloramphenicol, incubated at 30° C. for 3 days, and collected by scraping. Genomic DNA was extracted from the collected input and output cells using the cetyl trimethylammonium bromide (CTAB) method.

Quantitative PCR was performed with tag-specific primers using a qRT-PCR system (CFX96, Bio - Rad, Hercules, CA, USA). The STM score was calculated using the 2 ^-ΔΔCt method, and the relative change in genomic DNA amount was calculated. The average fold-change of the input versus output samples was calculated as a log score (log ₂ 2 ^{-(Ct, Target-Ct, Actin) output- (Ct, Target-Ct, Actin)input)} .

11. In vitro BBB-crossing and BBB-adhesion assay

Human brain microvascular endothelial cells (hCMEC/D3 cell line, Merck & Co., Kenilworth, NJ, USA) were cultured as follows based on a known method.

Briefly, 5 × 10 ⁴ hCMEC/D3 cells were prepared in EGM-2 medium (Lonza Group, Basel, Switzerland) and 8 μm porous membranes (BD) coated with collagen (Corning, Inc.) were prepared for the BBB-passage assay. Biosciences) and seeded in 12-well plates (BD Biosciences) for BBB-adhesion assay. The day after seeding, the medium was replaced with EGM-2 medium supplemented with 2.5% human serum and further cultured for 4 days. One day before C. neoformans inoculation, the medium was replaced with 0.5x diluted EGM-2 medium and the cells were maintained at 37°C and 5% CO ₂ . The integrity of the tight junction between cells was verified by confirming that a trans-endothelial electrical resistance (TEER) of ~200 Ω/cm ² was measured by an epithelial volt/ohm meter (EVOM2 device, World Precision Instruments) It became.

For the BBB-passage assay, 5 x 10 ⁵ cells of C. neoformans WT (H99), mpr1 Δ mutant and phosphatase deletion mutant were added to 500 μl of PBS and inoculated into a porous membrane. After culturing for 24 hours at 5% CO ₂ and 37° C., the number of cells passing through the porous membrane was measured by counting the CFU. Before and after cell inoculation, tight junction integrity was measured by TEER as described above. The BBB migration ratio was calculated by dividing the output CFU of each test strain by the input CFU of WT.

For the BBB adhesion assay, 100 μl PBS containing 5 x 10 ⁵ cells was inoculated onto a monolayer of hCMEC/D3 cells grown in a 12-well plate and incubated for 24 hours at 5% CO ₂ and 37°C. After culturing, the cultures were washed three times with PBS, and incubated with sterile distilled water at 37° C. for 30 minutes to burst and collect the host cells for CFU quantification. The BBB-adhesion ratio was calculated by dividing the attached CFU of each test strain by the CFU of WT C. neoformans.

12. Construction of database for C. neoformans phosphatase data

To facilitate access to phenotypic and genomic data of C. neoformans phosphatase mutant libraries, the Cryptococcus neoformans Phosphatase Phenome Database (http://phosphatase.cryptococcus.org) and the Cryptococcus neoformans Phenome Developed Gateway Database (http://www.cryptococcus.org/).

13. Statistical Analysis

Statistical analysis was performed using GraphPad Prism version 8. ANOVA with Bonferroni's multiple comparison test was used for capsule production analysis and expression analysis. Statistical analysis of pesticidal assays used the log-rank (Mantel-Cox) test. Statistical significance between ste50 Δ (positive control) and mutants in the murine STM assay was calculated by one-way ANOVA with Bonferroni's multiple comparison test.

Example 1: Identification of phosphatase gene of C. neoformans

Genes annotated as phosphatase were searched in the FungiDB C. neoformans (strain H99) genome database ( http://fungidb.org/fungidb ), and their protein sequences were analyzed for the presence of phosphatase-related domains. has been verified. Through this, 139 putative phosphatase genes of C. neoformans were selected. (See Table 2 below)

number H99 lD Gene name number H99 lD Gene name One CNAG_00057 FPP1 71 CNAG_03262 INM101 2 CNAG_01519 EEP1 72 CNAG_03296 SIW14 3 CNAG_02681 APH3 73 CNAG_03396 NPY1 4 CNAG_02944 APH1 74 CNAG_03491 NDX3 5 CNAG_03673 PPZ1 75 CNAG_03541 PTC7 6 CNAG_04354 XPP1 76 CNAG_03657 LDP1 7 CNAG_04453 IPC1 77 CNAG_03679 ACY1 8 CNAG_04782 PPU1 78 CNAG_03706 GLC7 9 CNAG_06115 APH4 79 CNAG_03765 TPS2 10 CNAG_06967 APH2 80 CNAG_03871 LTP1 11 CNAG_07629 EPP1 81 CNAG_03893 SDP102 12 CNAG_00076 NDX6 82 CNAG_03949 PHO13 13 CNAG_00146 PBY101 83 CNAG_03957 - 14 CNAG_00175 - 84 CNAG_03963 OCA1 15 CNAG_00182 VPS29 85 CNAG_03984 YCH1 16 CNAG_00263 PTP4 86 CNAG_03996 GDA1 17 CNAG_00265 IDI1 87 CNAG_04049 PPP5 18 CNAG_00316 HAM1 88 CNAG_04104 AKP3 19 CNAG_00390 YMR1 89 CNAG_04113 PTP3 20 CNAG_00408 PPH3 90 CNAG_04120 FCP1 21 CNAG_00427 PTC5 91 CNAG_04221 FBP26 22 CNAG_00498 CDC14 92 CNAG_04224 PSR1 23 CNAG_00500 PTC1 93 CNAG_04236 - 24 CNAG_00533 TIM50 94 CNAG_04268 APN2 25 CNAG_00554 ISC1 95 CNAG_04475 HPP3 26 CNAG_00790 DPP1 96 CNAG_04718 PCD101 27 CNAG_00810 HAD4 97 CNAG_04731 PCD102 28 CNAG_01054 SSU72 98 CNAG_04732 PCD1 29 CNAG_01177 NEM1 99 CNAG_04796 CNA1 30 CNAG_01203 YVH1 100 CNAG_04833 TEP1 31 CNAG_01254 OCA101 101 CNAG_04852 NDX4 32 CNAG_01314 PPP1 102 CNAG_05080 CWH43 33 CNAG_01346 HPP2 103 CNAG_05155 PTP2 34 CNAG_01357 SDP101 104 CNAG_05286 LDP2 35 CNAG_01412 PAH1 105 CNAG_05301 CRN1 36 CNAG_01436 SIT4 106 CNAG_05306 PTC701 37 CNAG_01496 PPT1 107 CNAG_05529 HAD5 38 CNAG_01498 AKP2 108 CNAG_05567 PPP3 39 CNAG_01518 PMU1 109 CNAG_05568 PRY1 40 CNAG_01532 INP52 110 CNAG_05617 GPI13 41 CNAG_01572 CDC25 111 CNAG_05639 PPS1 42 CNAG_01604 - 112 CNAG_05692 LCB3 43 CNAG_01744 HAD1 113 CNAG_05892 SHB17 44 CNAG_01823 INM1 114 CNAG_06064 PTP1 45 CNAG_01864 NDX5 115 CNAG_06065 SAC101 46 CNAG_01877 GUA1 116 CNAG_06080 SAC1 47 CNAG_01900 NDX1 117 CNAG_06122 HAD6 48 CNAG_01901 NDX2 118 CNAG_06132 HAD8 49 CNAG_02001 INP5203 119 CNAG_06141 DUT101 50 CNAG_02109 INP5201 120 CNAG_06232 HPP1 51 CNAG_02161 ASP1 121 CNAG_06236 NGL3 52 CNAG_02177 PPH22 122 CNAG_06251 PPP4 53 CNAG_02236 PPG1 123 CNAG_06418 PTC6 54 CNAG_02375 FIG4 124 CNAG_06499 DPP101 55 CNAG_02453 PTP5 125 CNAG_06549 CET1 56 CNAG_02470 PPP2 126 CNAG_06587 OCA2 57 CNAG_02487 PHS1 127 CNAG_06617 - 58 CNAG_02490 MRE11 128 CNAG_06647 CDC1 59 CNAG_02545 IPP1 129 CNAG_06698 HAD9 60 CNAG_02740 RPP1 130 CNAG_06900 - 61 CNAG_02772 PMU101 131 CNAG_06966 AKP1 62 CNAG_02861 HIS2 132 CNAG_06969 HAD7 63 CNAG_02878 GEP4 133 CNAG_07317 INP5202 64 CNAG_02986 YSA1 134 CNAG_07354 MET22 65 CNAG_03014 CAX4 135 CNAG_07372 OXK1 66 CNAG_03052 PTC2 136 CNAG_07434 HAD3 67 CNAG_03078 NPP1 137 CNAG_07547 GPM1 68 CNAG_03190 CCR4 138 CNAG_07692 YND1 69 CNAG_03202 CAC1 139 CNAG_07740 HAD2 70 CNAG_03222 DBR1

As a result of gene analysis of phosphatase of C. neoformans, the following three facts were found. (1) The degree of conservation of phosphatase in C. neoformans is lower than that of kinase but higher than that of transcription factor (TF), (2) There are only three types of tyrosine kinases in C. neoformans, which are insufficient. Despite containing 21 protein tyrosine phosphatases (PTPs), (3) C. neoformans contains a similar number of putative phosphatase genes as other fungi.

Example 2: Construction of C. neoformans phosphatase mutant library

Deletion mutants for the 139 putative phosphatase genes of Example 1 were prepared and their in vitro and in vivo phenotypic traits were analyzed.

Fifteen of the 139 putative phosphatase genes (PTP1, PTP2, YSA1, CNA1, CAC1, TPS2, CCR4, HAD1, EPP1, XPP1, APH1, ASP1, ISC1, PPG1, PPH3) have been functionally characterized in previous studies. In addition, in previous studies, two phosphatase genes with kinase domains ( OXK1 and FBP26 ) deletion strains were prepared for construction of kinase mutant libraries, and phosphatase genes with DNA binding domains ( APN2 ) were deleted for construction of TF mutant libraries. strains were made.

In addition to the 11 signature tag mutant strains constructed in previous studies (2 each for PTP1 , PTP2 , OXK1 , FBP26 , and APN2 ; 1 for CAC1 ), norseothricin containing a series of unique oligonucleotide signature tags We performed large-scale homologous recombination-based gene deletion using a nourseothricin resistance marker. In order to construct a high-quality phosphatase mutant library, two or more independent mutants were prepared for each gene, and the genotype was confirmed through diagnostic PCR and Southern blot analysis.

As a result, 219 mutant strains representing 109 phosphatases were newly prepared. Analysis of 230 mutant strains representing 114 phosphatases was performed along with 11 previously constructed mutant strains. (25 were excluded and the reason will be described later) In Table 3 below, 3741 and 2744 are controls, and 4341, 4342, 1429, 1430, 4288, 4289, 275, 277, 1704, 1705, and 42 were produced in previous studies. 11 strains, and the rest are 219 newly created mutant strains.

Gene ID Gene name Strain names
(YSB#) Genotypes Parents CNAG_07507 STE50 3741 MAT α ste50Δ::NAT-STM#234
(STM positive control) H99S CNAG_03670 IRE1 2744 MAT α ire1Δ::NAT-STM#169 (STM negative control) H99S CNAG_04221 FBP26 4341, 4342 MAT α fbp26Δ::NAT-STM#146 (previously constructed) H99S CNAG_04268 APN2 1429, 1430 MAT α apn2Δ::NAT-STM#102 (previously constructed) H99S CNAG_07372 OXK1 4288, 4289 MAT α oxk1Δ::NAT-STM#122
(previously constructed) H99S CNAG_05155 PTP2 275, 277 MAT α ptp2Δ::NAT-STM#184
(previously constructed) H99S CNAG_06064 PTP1 1704, 1705 MAT α ptp1Δ::NAT-STM#125 (previously constructed) H99S CNAG_03202 CAC1 42 MAT α cac1Δ::NAT-STM#159 (previously constructed) H99S CNAG_02986 YSA1 5013, 5014 MAT α ysa1Δ::NAT-STM#116 H99S CNAG_00057 FPP1 4596, 4597 MAT α fpp1Δ::NAT-STM#184 H99S CNAG_01519 EEP1 4659, 4662 MAT α eep1Δ::NAT-STM#191 H99S CNAG_02681 APH3 4051, 4052 MAT α aph3Δ::NAT-STM#150 H99S CNAG_02944 APH1 3478, 3479 MAT α aph1Δ::NAT-STM#191 H99S CNAG_03673 PPZ1 5788, 5789 MAT α ppz1Δ::NAT-STM#295 H99S CNAG_04354 XPP1 5941, 5942, 6628 MAT α xpp1Δ::NAT-STM#234 H99S CNAG_04782 PPU1 6630, 6631 MAT α ppu1Δ::NAT-STM#212 H99S CNAG_06967 APH2 3484, 3485 MAT α aph2Δ::NAT-STM#159 H99S CNAG_07629 EPP1 4922, 4924 MAT α epp1Δ::NAT-STM#240 H99S CNAG_00076 NDX6 5140, 5141 MAT α ndx6Δ::NAT-STM#159 H99S CNAG_00146 PBY101 5153, 5154 MAT α pby101Δ::NAT-STM#5 H99S CNAG_00182 VPS29 4881, 4882 MAT α vps29Δ::NAT-STM#295 H99S CNAG_00263 PTP4 5387, 5388 MAT α ptp4Δ::NAT-STM#123 H99S CNAG_00316 HAM1 5184, 5186 MAT α ham1Δ::NAT-STM#212 H99S CNAG_00390 YMR1 4262, 4554 MAT α ymr1Δ::NAT-STM#5 H99S CNAG_00408 PPH3 3776, 3778 MAT α pph3Δ::NAT-STM#146 H99S CNAG_00427 PTC5 5943, 5944 MAT α ptc5Δ::NAT-STM#43 H99S CNAG_00498 CDC14 3620, 5359 MAT α cdc14Δ::NAT-STM#116 H99S CNAG_00500 PTC1 289, 290 MAT α ptc1Δ::NAT-STM#146 H99S CNAG_00554 ISC1 4264, 4265 MAT α isc1Δ::NAT-STM#43 H99S CNAG_00810 HAD4 4194, 4197 MAT α had4Δ::NAT-STM#116 H99S CNAG_01054 SSU72 4242, 4243 MAT α ssu72Δ::NAT-STM#119 H99S CNAG_01177 NEM1 4771, 4772 MAT α nem1Δ::NAT-STM#177 H99S CNAG_01203 YVH1 4590, 4591 MAT α yvh1Δ::NAT-STM#5 H99S CNAG_01254 OCA101 4075, 4077 MAT α oca101Δ::NAT-STM#210 H99S CNAG_01314 PPP1 5058, 5059 MAT α ppp1Δ::NAT-STM#184 H99S CNAG_01346 HPP2 4728, 4730 MAT α hpp2Δ::NAT-STM#204 H99S CNAG_01357 SDP101 3613, 3614 MAT α sdp101Δ::NAT-STM#6 H99S CNAG_01436 SIT4 4094, 4095 MAT α sit4Δ::NAT-STM#232 H99S CNAG_01496 PPT1 5554, 5555 MAT α ppt1Δ::NAT-STM#184 H99S CNAG_01498 AKP2 5064, 5069 MAT α akp2Δ::NAT-STM#6 H99S CNAG_01518 PMU1 4026, 4027 MAT α pmu1Δ::NAT-STM#205 H99S CNAG_01532 INP52 4793, 4794 MAT α inp52Δ::NAT-STM#201 H99S CNAG_01744 HAD1 4665, 4666 MAT α had1Δ::NAT-STM#204 H99S CNAG_01823 INM1 4608, 4609 MAT α inm1Δ::NAT-STM#191 H99S CNAG_01864 NDX5 5466, 5467 MAT α ndx5Δ::NAT-STM#150 H99S CNAG_01877 GUA1 6161, 6163 MAT α gua1Δ::NAT-STM#213 H99S CNAG_01900 NDX1 5248, 5249 MAT α ndx1Δ::NAT-STM#210 H99S CNAG_01901 NDX2 5504, 5505 MAT α ndx2Δ::NAT-STM#211 H99S CNAG_02001 INP5203 4692, 4693 MAT α inp5203Δ::NAT-STM#205 H99S CNAG_02109 INP5201 4607, 4929 MAT α inp5201Δ::NAT-STM#122 H99S CNAG_02161 ASP1 5250, 5251 MAT α asp1Δ::NAT-STM#240 H99S CNAG_02236 PPG1 5772, 5940 MAT α ppg1Δ::NAT-STM#123 H99S CNAG_02375 FIG4 4574, 4575 MAT α fig4Δ::NAT-STM#150 H99S CNAG_02453 PTP5 5469, 5470 MAT α ptp5Δ::NAT-STM#191 H99S CNAG_02470 PPP2 4819, 4822 MAT α ppp2Δ::NAT-STM#290 H99S CNAG_02487 PHS1 4022, 4023 MAT α phs1Δ::NAT-STM#225 H99S CNAG_02490 MRE11 4694, 5050 MAT α mre11Δ::NAT-STM#242 H99S CNAG_02772 PMU101 4670, 4671 MAT α pmu101Δ::NAT-STM#208 H99S CNAG_03014 CAX4 4790, 4791 MAT αcax4Δ::NAT-STM#211 H99S CNAG_03052 PTC2 1772, 1773 MAT α ptc2Δ::NAT-STM#224 H99S CNAG_03078 NPP1 5019, 5021 MAT α npp1Δ::NAT-STM#119 H99S CNAG_03190 CCR4 5591, 5594 MAT α ccr4Δ::NAT-STM#210 H99S CNAG_03202 CAC1 5650 MAT α cac1Δ::NAT-STM#159 H99S CNAG_03222 DBR1 4963, 4964 MAT α dbr1Δ::NAT-STM#288 H99S CNAG_03262 INM101 4529, 4530 MAT α inm101Δ::NAT-STM#201 H99S CNAG_03296 SIW14 4570, 4571 MAT α siw14Δ::NAT-STM#225 H99S CNAG_03396 NPY1 5105, 5107 MAT α npy1Δ::NAT-STM#122 H99S CNAG_03491 NDX3 5192, 5193 MAT α ndx3Δ::NAT-STM#201 H99S CNAG_03541 PTC7 1860, 1861 MAT α ptc71Δ::NAT-STM#53 H99S CNAG_03657 LDP1 4382, 4383 MAT α ldp1Δ::NAT-STM#231 H99S CNAG_03679 ACY1 5838, 5839 MAT α acy1Δ::NAT-STM#43 H99S CNAG_03765 TPS2 4373, 4433 MAT α tps2Δ::NAT-STM#232 H99S CNAG_03871 LTP1 4044, 4045 MAT α ltp1Δ::NAT-STM#230 H99S CNAG_03893 SDP102 3467, 3468 MAT α sdp102Δ::NAT-STM#119 H99S CNAG_03949 PHO13 4374, 4376 MAT α pho13Δ::NAT-STM#234 H99S CNAG_03963 OCA1 4054, 4055 MAT α oca1Δ::NAT-STM#230 H99S CNAG_03984 YCH1 4637, 4638 MAT α ych1Δ::NAT-STM#208 H99S CNAG_03996 GDA1 4750, 5821 MAT α gda1Δ::NAT-STM#6 H99S CNAG_04049 PPP5 4298, 4299 MAT α ppp5Δ::NAT-STM#288 H99S CNAG_04104 AKP3 5484, 5485 MAT α akp3Δ::NAT-STM#58 H99S CNAG_04113 PTP3 4680, 4754 MAT α ptp3Δ::NAT-STM#231 H99S CNAG_04224 PSR1 5003, 5005 MAT α psr1Δ::NAT-STM#125 H99S CNAG_04475 HPP3 4003, 4004 MAT α hpp3Δ::NAT-STM#211 H99S CNAG_04718 PCD101 5416, 5417 MAT α pcd101Δ::NAT-STM#146 H99S CNAG_04731 PCD102 5422, 5424 MAT α pcd102Δ::NAT-STM#125 H99S CNAG_04732 PCD1 5045, 5053 MAT α pcd1Δ::NAT-STM#123 H99S CNAG_04796 CNA1 4067, 4068 MAT α cna1Δ::NAT-STM#290 H99S CNAG_04833 TEP1 4622, 4623 MAT α tep1Δ::NAT-STM#205 H99S CNAG_04852 NDX4 5046, 5047 MAT α ndx4Δ::NAT-STM#204 H99S CNAG_05080 CWH43 4508, 4509 MAT α cwh43Δ::NAT-STM#213 H99S CNAG_05286 LDP2 4721, 4722 MAT α ldp2Δ::NAT-STM#218 H99S CNAG_05301 CRN1 5095, 5096 MAT α crn1Δ::NAT-STM#218 H99S CNAG_05306 PTC701 1867, 1868 MAT α ptc701Δ::NAT-STM#58 H99S CNAG_05529 HAD5 5098, 5099 MAT α had5Δ::NAT-STM#205 H99S CNAG_05567 PPP3 4396, 4399 MAT α ppp3Δ::NAT-STM#288 H99S CNAG_05568 PRY1 5101, 5102 MAT α pry1Δ::NAT-STM#211 H99S CNAG_05692 LCB3 4755, 5051, 5052 MAT α lcb3Δ::NAT-STM#219 H99S CNAG_05892 SHB17 4060, 4061 MAT α shb17Δ::NAT-STM#212 H99S CNAG_06065 SAC101 4470, 4718 MAT α sac101Δ::NAT-STM#220 H99S CNAG_06122 HAD6 5476, 5477 MAT α had6Δ::NAT-STM#219 H99S CNAG_06132 HAD8 4464, 4475 MAT α had8Δ::NAT-STM#242 H99S CNAG_06141 DUT101 5112, 5128 MAT α dut101Δ::NAT-STM#208 H99S CNAG_06232 HPP1 4092, 4093 MAT α hpp1Δ::NAT-STM#213 H99S CNAG_06236 NGL3 4459, 4472 MAT α ngl3Δ::NAT-STM#273 H99S CNAG_06251 PPP4 4784, 4785 MAT α ppp4Δ::NAT-STM#296 H99S CNAG_06418 PTC6 1875, 1876 MAT α ptc6Δ::NAT-STM#102 H99S CNAG_06499 DPP101 4479, 4480 MAT α dpp101Δ::NAT-STM#290 H99S CNAG_06587 OCA2 4841, 4842 MAT α oca2Δ::NAT-STM#231 H99S CNAG_06647 CDC1 5542, 5574 MAT α cdc1Δ::NAT-STM#296 H99S CNAG_06698 HAD9 4485, 4486 MAT α had9Δ::NAT-STM#58 H99S CNAG_06966 AKP1 4892, 4893 MAT α akp1Δ::NAT-STM#150 H99S CNAG_06969 HAD7 4688, 4946 MAT α had7Δ::NAT-STM#220 H99S CNAG_07317 INP5202 4511, 4538 MAT α inp5202Δ::NAT-STM#102 H99S CNAG_07434 HAD3 4624, 4626 MAT α had3Δ::NAT-STM#116 H99S CNAG_07547 GPM1 4089, 4090 MAT α gpm1Δ::NAT-STM#219 H99S CNAG_07692 YND1 4856, 6157 MAT α ynd1Δ::NAT-STM#119 H99S CNAG_07740 HAD2 4506, 4507 MAT α had2Δ::NAT-STM#123 H99S CNAG_00508 VPS17 5724 MAT α vps17Δ::NAT-STM#220 H99S CNAG_01315 VPS5 5683, 5684 MAT α vps5Δ::NAT-STM#221 H99S CNAG_01426 VPS26 5671, 5672 MAT α vps26Δ::NAT-STM#5 H99S CNAG_01837 VPS35 5615, 5616 MAT α vps35Δ::NAT-STM#210 H99S Double EN OCA101, OCA1 6634, 6635 MAT α oca101Δ::NAT oca1Δ::NEO YSB4075

Disruption strategies, primer sequences, southern blot results, and mutation data can be found in the Cryptococcus neoformans Phosphatase Phenome Database ( http://phosphatase.cryptococcus.org ) constructed by the present inventors.

01572/ CDC25 , 06080/ SAC1 , 04120/ FCP1 , 04453/ IPC1 , 06549/ CET1 , 03706/ GLC7 , 00533/ TIM50 , 00265/ IDI1 , 02545/ IPP1 , 0/ RPP1 02861/ HIS2 , 07354/ MET22 , 01412/ PAH1 , 05617/ GPI13 , 05639/ PPS1 , 00790/ DPP1 , 03957/-, 02177/ PPH22 , 02878/ GEP4 , 01604/-, 06900/07, 1/5,60/-07 -, 06115/ APH4 , 04236/-), viable transformants were not obtained or only potential aneuploidy mutants possessing both wild-type and mutant alleles were obtained It became. This suggests that the remaining 25 genes may be essential for survival.

Example 3: Phosphatome phenotype and in vivo expression profiling

Phenotypes were examined under 30 types of in vitro conditions using 230 C. neoformans phosphatase deletion mutants for 114 genes prepared in Example 2. 30 types of conditions were tested for growth at various temperatures (25, 30, 37 and 39°C), mating efficiency, production of virulence factors (capsule, melanin and urease), stress response (osmotic/cationic salts, oxidation, genotoxicity, ER, cell membrane). / wall and heavy metal stress) and antifungal drug susceptibility. (See Table 4 below)

Abb. Description Tested condition One Growth at different temperatures 25 incubated at 25 ^o C Cells were incubated at 25 ^o C for 1-6 days. 2 30 incubated at 30 ^o C Cells were incubated at 30 ^o C for 1-6 days. 3 37 incubated at 37 ^o C Cells were incubated at 37 ^o C for 1-6 days. 4 39 incubated at 39 ^o C Cells were incubated at 39 ^o C for 1-6 days. 5 Mating MAT Mating Cells were incubated with opposite mating type cells onto the V8 media in the dark for 15 days. 6 Virulence factors MEL Melanin Cells were spotted onto the Niger seed media containing glucose and further incubated at 37C for 1-4 days. 7 CAP Capsule Cells were incubated onto the solid-agared DME media and relative packed cell volume was measured 8 URE Urease Cells were spotted onto the Christensen's agar media and further incubated at 30 ^o C for 1-4 days. 9 Osmotic/cation salt stress NCR NaCl treatment under glucose-rich conditions Cells were spotted onto the YPD media containing 1.5 M NaCl for 1-6 days. 10 KCR KCl treatment under glucose-rich condition Cells were spotted onto the YPD media containing 1.5 M KCl for 1-6 days. 11 SBR Sorbitol treatment under glucose-rich condition Cells were spotted onto the YPD media containing 2 M sorbitol for 1-6 days. 12 NCS NaCl treatment under glucose-starved condition Cells were spotted onto the YP media containing 1 M NaCl for 1-6 days. 13 KCS KCl treatment under glucose-starved condition Cells were spotted onto the YP media containing 1 M KCl for 1-6 days. 14 SBS Sorbitol treatment for underglucose-starved condition Cells were spotted onto the YP media containing 2 M sorbitol for 1-6 days. 15 Oxidative stress HPX Hydrogen peroxide Cells were spotted onto the YPD media containing 2.5 to 3.5 mM concentration of hydrogen peroxide for 1-6 days. 16 TBH Tert-butyl hydroperoxide Cells were spotted onto the YPD media containing 0.6 to 0.8 mM concentration of tert-butyl hydroperoxide for 1-6 days. 17 MD Menadione Cells were spotted onto the YPD media containing 0.01 to 0.03 mM concentration of menadione for 1-6 days. 18 DIA Diamide Cells were spotted onto the YPD media containing 2-3.5 mM concentration of diamide for 1-6 days. 19 Genotoxic stress MMS Methyl methanesulfonate Cells were spotted onto the YPD media containing 0.02 to 0.06% MMS for 1-6 days. 20 HU Hydroxyurea Cells were spotted onto the YPD media containing 90 to 110 mM concentration of HU for 1-6 days. 21 ER
Stress TM Tunicamycin Cells were spotted onto the YPD media containing 0.2 to 0.4 μg/ml concentration of TM for 1-6 days. 22 DTT Dithiothreitol Cells were spotted onto the YPD media containing 15 to 20 mM concentration of DTT for 1-6 days. 23 Heavy metal stress CDS CdSO ₄ Cells were spotted onto the YPD media containing 20 to 35 µM concentration of for CdSO4 1-6 days. 24 Cell wall/membrane stress CR Congo - red Cells were spotted onto the YPD media containing 0.8 to 1% congo - red for 1-6 days. 25 CFW Calcofluor white Cells were spotted onto the YPD media containing 3-5 mg/ml concentration of CFW for 1-6 days. 26 SDS Sodium dodecyl sulfate Cells were spotted onto the YPD media containing 0.03 to 0.05% SDS for 1-6 days. 27 Antifungal drug susceptibility 5FC Flucytosine Cells were spotted onto the YPD media containing 300 to 500 μg/ml concentration of flucytosine for 1-6 days. 28 AMB Amphotericin B Cells were spotted onto the YPD media containing 0.8 to 1.8 μg/ml concentration of Amphotericin B for 1-6 days. 29 FCZ Fluconazole Cells were spotted onto the YPD media containing 10-18 μg/ml concentration of fluconazole for 1-6 days. 30 FDX Fludioxonil Cells were spotted onto the YPD media containing 0.5 to 5 μg/ml concentration of fludioxonil for 1-6 days.

Figure 1 shows in vitro phenotypic changes and in vivo expression profiling for each cluster in vitro by phosphatase deletion of C. neoformans. Tables 5 to 10 below show data for 30 phenotypes in detail by cluster. In Figure 1 and Tables 5 to 10, each value is -3: strong reduction / sensitivity, -2: moderately reduction / sensitivity, -1: weak reduction / sensitivity, 0: similar to wild type, +1 weak Increase/resistance, +2: Slight increase/resistance, +3: Strong increase/resistance.

Table 5 below shows the results of confirming the effect of deletion of each gene on growth at 25, 30, 37 and 39 ° C.

Growth at different temperatures H99 lD Name 25 30 37 39 00057 FPP1 0 0 0 0 00076 NDX6 0 0 0 0 00146 PBY101 0 0 0 One 00182 VPS29 0 0 0 0 00263 PTP4 0 0 0 0 00316 HAM1 0 0 0 0 00390 YMR1 0 0 0 0 00408 PPH3 0 0 0 0 00427 PTC5 0 0 0 0 00498 CDC14 0 0 0 0 00500 PTC1 0 0 0 0 00554 ISC1 0 0 0 0 00810 HAD4 0 0 0 0 01054 SSU72 0 0 -One -One 01177 NEM1 0 -One -2 -3 01203 YVH1 0 -2 -One -One 01254 OCA101 -One -One 0 0 01314 PPP1 0 0 0 0 01346 HPP2 0 0 0 0 01357 SDP101 0 0 0 0 01436 SIT4 0 0 0 0 01496 PPT1 0 0 0 0 01498 AKP2 0 0 0 0 01518 PMU1 0 0 0 0 01519 EEP1 0 0 0 0 01532 INP52 0 0 0 0 01744 HAD1 -One 0 0 0 01823 INM1 0 0 0 0 01864 NDX5 0 0 0 0 01877 GUA1 0 -2 -One -2 01900 NDX1 0 0 0 0 01901 NDX2 0 0 0 0 02001 INP5203 0 0 0 0 02109 INP5201 0 -One -One -2 02161 ASP1 0 0 0 0 02236 PPG1 0 -2 -3 -3 02375 FIG4 0 0 0 -2 02453 PTP5 0 0 0 0 02470 PPP2 0 0 0 -One 02487 PHS1 0 0 -One -One 02490 MRE11 0 0 -2 -2 02681 APH3 0 0 0 0 02772 PMU101 0 0 0 0 02944 APH1 0 0 0 0 02986 YSA1 0 0 0 0 03014 CAX4 0 0 0 0 03052 PTC2 -One -One 0 0 03078 NPP1 0 0 0 0 03190 CCR4 0 -One -2 -3 03202 CAC1 0 0 0 0 03222 DBR1 0 -2 -One -2 03262 INM101 0 0 0 0 03296 SIW14 0 -One -One -One 03396 NPY1 0 0 0 0 03491 NDX3 0 0 0 0 03541 PTC7 0 0 0 0 03657 LDP1 0 0 0 0 03673 PPZ1 0 0 0 0 03679 ACY1 0 0 0 0 03765 TPS2 0 0 -3 -3 03871 LTP1 0 0 0 0 03893 SDP102 0 0 0 0 03949 PHO13 0 0 0 0 03963 OCA1 0 0 0 0 03984 YCH1 0 0 0 0 03996 GDA1 0 0 0 0 04049 PPP5 0 0 0 0 04104 AKP3 0 0 0 0 04113 PTP3 0 0 0 0 04221 FBP26 -One -One -One -One 04224 PSR1 0 0 0 0 04268 APN2 0 0 0 -One 04354 XPP1 0 0 0 0 04475 HPP3 0 0 0 0 04718 PCD101 0 0 0 0 04731 PCD102 0 0 0 0 04732 PCD1 0 0 0 0 04782 PPU1 0 0 0 0 04796 CNA1 0 0 -2 -3 04833 TEP1 0 0 0 0 04852 NDX4 0 0 0 0 05080 CWH43 0 0 0 0 05155 PTP2 0 -One 0 0 05286 LDP2 0 0 0 0 05301 CRN1 0 0 0 0 05306 PTC701 0 0 0 0 05529 HAD5 0 0 0 0 05567 PPP3 0 0 0 0 05568 PRY1 0 0 0 0 05892 SHB17 0 0 0 0 06064 PTP1 0 0 0 0 06065 SAC101 0 0 0 0 06122 HAD6 0 0 0 0 06132 HAD8 0 0 0 0 06141 DUT101 0 0 0 0 06232 HPP1 0 0 0 0 06236 NGL3 0 0 0 0 06251 PPP4 0 0 0 0 06418 PTC6 0 0 0 0 06499 DPP101 0 0 0 2 06587 OCA2 0 0 0 0 06647 CDC1 0 0 0 0 06698 HAD9 0 0 0 0 06966 AKP1 0 0 0 0 06967 APH2 0 0 0 0 06969 HAD7 0 0 0 0 07317 INP5202 0 0 0 0 07372 OXK1 0 0 -One 0 07434 HAD3 0 0 0 0 07547 GPM1 0 0 0 0 07629 EPP1 0 0 0 0 07692 YND1 0 0 0 0 07740 HAD2 0 0 0 0

Table 6 below shows the effect of deletion of each gene on mating efficiency and virulence factors.

Mating Virulence factors H99 lD Name MAT MEL CAP URE 00057 FPP1 0 0 0 3 00076 NDX6 0 0 0 0 00146 PBY101 -2 0 0 0 00182 VPS29 0 -One 0 2 00263 PTP4 0 0 0 0 00316 HAM1 0 0 0 0 00390 YMR1 0 0 0 0 00408 PPH3 0 0 0 0 00427 PTC5 0 0 0 0 00498 CDC14 0 0 0 0 00500 PTC1 One 0 0 0 00554 ISC1 0 0 0 0 00810 HAD4 0 0 0 0 01054 SSU72 -3 -One 0 0 01177 NEM1 -3 -2 One 3 01203 YVH1 -2 -One 0 -3 01254 OCA101 -3 0 0 0 01314 PPP1 0 0 0 0 01346 HPP2 0 0 -2 0 01357 SDP101 0 0 0 0 01436 SIT4 0 0 2 0 01496 PPT1 0 0 0 0 01498 AKP2 0 0 0 0 01518 PMU1 0 0 0 0 01519 EEP1 0 0 0 0 01532 INP52 0 0 0 0 01744 HAD1 -2 0 0 0 01823 INM1 0 0 0 0 01864 NDX5 0 0 -One 0 01877 GUA1 -3 -3 0 -3 01900 NDX1 0 0 0 0 01901 NDX2 0 0 0 0 02001 INP5203 0 0 0 0 02109 INP5201 -3 -3 3 3 02161 ASP1 0 0 0 0 02236 PPG1 -3 -3 0 3 02375 FIG4 0 0 One 0 02453 PTP5 0 0 0 -2 02470 PPP2 0 0 0 0 02487 PHS1 0 0 3 0 02490 MRE11 0 -One 0 0 02681 APH3 0 0 0 0 02772 PMU101 0 0 0 0 02944 APH1 0 0 0 0 02986 YSA1 0 0 0 0 03014 CAX4 0 One 0 0 03052 PTC2 -3 -3 0 0 03078 NPP1 0 0 0 0 03190 CCR4 -2 -One 2 3 03202 CAC1 -3 -3 -3 One 03222 DBR1 -3 -3 3 3 03262 INM101 0 0 0 0 03296 SIW14 -2 One 0 -One 03396 NPY1 0 0 0 0 03491 NDX3 0 0 0 0 03541 PTC7 0 0 0 0 03657 LDP1 0 0 0 0 03673 PPZ1 0 0 0 0 03679 ACY1 0 0 0 0 03765 TPS2 0 -2 0 One 03871 LTP1 0 0 0 0 03893 SDP102 0 0 -One 0 03949 PHO13 0 0 0 0 03963 OCA1 0 0 -One 0 03984 YCH1 0 0 0 0 03996 GDA1 -One 0 0 0 04049 PPP5 0 0 -One 0 04104 AKP3 0 0 0 0 04113 PTP3 0 0 0 0 04221 FBP26 -3 -3 0 0 04224 PSR1 0 -One -2 0 04268 APN2 0 0 0 0 04354 XPP1 0 -One 0 2 04475 HPP3 0 0 0 0 04718 PCD101 0 0 0 0 04731 PCD102 0 0 One 0 04732 PCD1 0 0 0 0 04782 PPU1 0 0 0 0 04796 CNA1 -2 -3 2 2 04833 TEP1 0 0 0 0 04852 NDX4 0 0 0 0 05080 CWH43 0 0 -One 0 05155 PTP2 -3 -3 -2 0 05286 LDP2 0 0 0 0 05301 CRN1 2 0 0 0 05306 PTC701 0 0 0 0 05529 HAD5 0 0 0 0 05567 PPP3 0 0 0 0 05568 PRY1 0 0 0 0 05892 SHB17 0 0 0 0 06064 PTP1 0 0 0 0 06065 SAC101 0 0 0 0 06122 HAD6 0 0 0 0 06132 HAD8 0 0 0 0 06141 DUT101 0 0 0 0 06232 HPP1 0 0 0 0 06236 NGL3 0 0 -One 0 06251 PPP4 0 0 0 0 06418 PTC6 -One 0 0 0 06499 DPP101 0 0 0 0 06587 OCA2 0 0 0 0 06647 CDC1 0 0 2 0 06698 HAD9 0 0 0 0 06966 AKP1 0 0 0 0 06967 APH2 0 0 0 0 06969 HAD7 0 0 0 0 07317 INP5202 0 0 0 0 07372 OXK1 0 -2 0 0 07434 HAD3 0 0 0 0 07547 GPM1 0 0 0 0 07629 EPP1 0 0 0 0 07692 YND1 0 0 -2 0 07740 HAD2 0 0 0 0

Table 7 below shows the results of confirming the effect of deletion of each gene on osmotic stress.

Osmotic/cation salt stress H99 lD Name KCR NCR SBR KCS NCS SBS 00057 FPP1 0 0 0 0 0 0 00076 NDX6 0 0 0 0 0 0 00146 PBY101 0 0 0 0 0 0 00182 VPS29 0 0 0 0 0 0 00263 PTP4 0 0 0 0 0 0 00316 HAM1 0 0 0 0 0 0 00390 YMR1 0 0 0 0 0 0 00408 PPH3 0 0 0 0 0 0 00427 PTC5 0 0 0 0 0 0 00498 CDC14 0 0 0 0 0 0 00500 PTC1 0 0 0 0 0 0 00554 ISC1 0 -3 0 0 -3 0 00810 HAD4 0 0 0 0 0 0 01054 SSU72 -2 -3 -2 -One -2 -One 01177 NEM1 -One -3 -One -One -One -One 01203 YVH1 0 0 0 0 0 0 01254 OCA101 0 0 0 0 0 0 01314 PPP1 0 0 0 0 0 0 01346 HPP2 0 0 0 0 0 0 01357 SDP101 0 0 0 0 0 0 01436 SIT4 0 0 0 0 0 0 01496 PPT1 0 0 0 0 0 0 01498 AKP2 0 0 0 0 0 0 01518 PMU1 0 0 0 0 0 0 01519 EEP1 0 0 0 0 0 0 01532 INP52 0 0 0 0 0 0 01744 HAD1 -One -One -One 0 0 0 01823 INM1 0 0 0 0 0 0 01864 NDX5 0 0 0 0 0 0 01877 GUA1 -3 -3 -3 -2 -2 -2 01900 NDX1 0 0 0 0 0 0 01901 NDX2 0 0 0 0 0 0 02001 INP5203 0 0 0 0 0 0 02109 INP5201 -2 -3 -One -2 -2 -2 02161 ASP1 0 0 0 0 0 0 02236 PPG1 -One -3 -One 0 -3 0 02375 FIG4 0 0 0 0 0 0 02453 PTP5 0 0 0 0 0 0 02470 PPP2 0 0 0 0 0 0 02487 PHS1 -2 -2 0 -2 -2 0 02490 MRE11 0 0 0 0 0 0 02681 APH3 0 0 0 0 0 0 02772 PMU101 0 0 0 0 0 0 02944 APH1 0 0 0 0 0 0 02986 YSA1 0 0 0 0 0 0 03014 CAX4 0 0 0 0 0 0 03052 PTC2 0 0 -One 0 0 0 03078 NPP1 0 0 0 0 0 0 03190 CCR4 0 0 0 0 0 0 03202 CAC1 0 0 0 0 0 0 03222 DBR1 -2 -2 -2 -2 -3 -2 03262 INM101 0 0 0 0 0 0 03296 SIW14 -One -One 0 -One -One 0 03396 NPY1 0 0 0 0 0 0 03491 NDX3 0 0 0 0 0 0 03541 PTC7 0 0 0 0 0 0 03657 LDP1 0 0 0 0 0 0 03673 PPZ1 0 -2 0 0 0 0 03679 ACY1 0 0 0 0 0 0 03765 TPS2 0 0 0 0 0 0 03871 LTP1 0 0 0 0 0 0 03893 SDP102 0 0 0 0 0 0 03949 PHO13 0 0 0 0 0 0 03963 OCA1 0 0 0 0 0 0 03984 YCH1 0 0 0 0 0 0 03996 GDA1 0 -2 -2 0 -3 -One 04049 PPP5 0 0 0 0 0 0 04104 AKP3 0 0 0 0 0 0 04113 PTP3 0 0 0 0 0 0 04221 FBP26 0 -One -One 0 -One -One 04224 PSR1 0 0 0 0 0 0 04268 APN2 0 0 0 0 0 0 04354 XPP1 0 0 0 0 0 0 04475 HPP3 0 0 0 0 0 0 04718 PCD101 0 0 0 0 0 0 04731 PCD102 0 0 0 0 0 0 04732 PCD1 0 0 0 0 0 0 04782 PPU1 0 0 0 0 0 0 04796 CNA1 -2 -3 -2 -One -2 -2 04833 TEP1 0 0 0 0 0 0 04852 NDX4 0 0 0 0 0 0 05080 CWH43 0 0 0 0 0 0 05155 PTP2 -One -One -One -One -One -One 05286 LDP2 0 0 0 0 0 0 05301 CRN1 0 0 0 0 0 0 05306 PTC701 0 0 0 0 0 0 05529 HAD5 0 0 0 0 0 0 05567 PPP3 0 0 0 0 0 0 05568 PRY1 0 0 0 0 0 0 05892 SHB17 0 0 0 0 0 0 06064 PTP1 0 0 0 0 0 0 06065 SAC101 0 0 0 0 0 0 06122 HAD6 0 0 0 0 0 0 06132 HAD8 0 0 0 0 0 0 06141 DUT101 0 0 0 0 0 0 06232 HPP1 0 0 0 0 0 0 06236 NGL3 0 0 0 0 0 0 06251 PPP4 0 0 0 0 0 0 06418 PTC6 0 -2 0 0 0 0 06499 DPP101 0 0 0 0 0 0 06587 OCA2 0 0 0 0 0 0 06647 CDC1 0 0 0 0 0 0 06698 HAD9 0 0 0 0 0 0 06966 AKP1 0 0 0 0 0 0 06967 APH2 0 0 0 0 0 0 06969 HAD7 0 0 0 0 0 0 07317 INP5202 0 0 0 0 0 0 07372 OXK1 One 0 0 0 0 0 07434 HAD3 0 0 0 0 0 0 07547 GPM1 0 0 0 0 0 0 07629 EPP1 0 0 0 0 0 0 07692 YND1 0 0 0 0 0 0 07740 HAD2 0 0 0 0 0 0

Table 8 below shows the results of confirming the effect of deletion of each gene on oxidative stress.

Oxidative stress H99 lD Name DIA HPX MD TBH 00057 FPP1 0 0 0 0 00076 NDX6 0 0 0 0 00146 PBY101 0 0 0 0 00182 VPS29 0 0 0 0 00263 PTP4 0 0 0 0 00316 HAM1 0 0 0 0 00390 YMR1 0 0 0 0 00408 PPH3 -One 0 0 0 00427 PTC5 0 0 0 0 00498 CDC14 0 0 0 0 00500 PTC1 0 0 0 0 00554 ISC1 -2 0 0 -One 00810 HAD4 0 0 0 0 01054 SSU72 -2 -One -One -One 01177 NEM1 -2 -2 -One -3 01203 YVH1 0 -One -One -One 01254 OCA101 0 0 One -One 01314 PPP1 0 0 0 0 01346 HPP2 0 0 0 0 01357 SDP101 0 0 0 0 01436 SIT4 -One -One -One -One 01496 PPT1 -One 0 0 0 01498 AKP2 0 0 0 0 01518 PMU1 0 0 0 0 01519 EEP1 0 0 0 0 01532 INP52 0 0 0 0 01744 HAD1 -One 0 -One 0 01823 INM1 0 0 0 0 01864 NDX5 0 0 0 0 01877 GUA1 0 -2 -One -One 01900 NDX1 0 0 0 0 01901 NDX2 0 0 0 0 02001 INP5203 0 0 0 0 02109 INP5201 -2 -2 -2 -2 02161 ASP1 0 0 0 0 02236 PPG1 -3 -One -One -One 02375 FIG4 0 0 0 0 02453 PTP5 0 -2 0 0 02470 PPP2 0 0 0 0 02487 PHS1 -3 -One 0 -3 02490 MRE11 -One -2 0 -One 02681 APH3 0 0 0 0 02772 PMU101 0 0 0 0 02944 APH1 0 0 0 0 02986 YSA1 0 0 0 0 03014 CAX4 0 0 0 0 03052 PTC2 -One 0 -One -One 03078 NPP1 0 0 0 0 03190 CCR4 -One -One 0 -2 03202 CAC1 One -One 0 -One 03222 DBR1 -One -One -One -2 03262 INM101 0 0 0 0 03296 SIW14 -One 0 0 -2 03396 NPY1 0 0 0 0 03491 NDX3 0 0 0 0 03541 PTC7 0 0 0 0 03657 LDP1 0 0 0 0 03673 PPZ1 One -2 0 -One 03679 ACY1 0 0 0 0 03765 TPS2 0 0 0 One 03871 LTP1 0 0 0 0 03893 SDP102 0 0 0 0 03949 PHO13 0 0 0 0 03963 OCA1 -One 0 0 -2 03984 YCH1 0 0 0 0 03996 GDA1 -One 0 0 0 04049 PPP5 0 0 0 0 04104 AKP3 0 0 0 0 04113 PTP3 0 0 0 0 04221 FBP26 -3 0 -2 0 04224 PSR1 -One 0 0 0 04268 APN2 0 0 0 0 04354 XPP1 0 0 0 0 04475 HPP3 0 0 0 0 04718 PCD101 0 0 0 0 04731 PCD102 0 0 0 0 04732 PCD1 0 0 0 0 04782 PPU1 0 0 0 0 04796 CNA1 -3 0 0 -One 04833 TEP1 0 0 0 0 04852 NDX4 0 0 0 0 05080 CWH43 0 0 0 0 05155 PTP2 -3 0 -One -One 05286 LDP2 0 0 0 0 05301 CRN1 0 0 0 0 05306 PTC701 0 0 0 0 05529 HAD5 0 0 0 0 05567 PPP3 0 0 0 0 05568 PRY1 0 0 0 0 05892 SHB17 0 0 0 0 06064 PTP1 0 0 0 0 06065 SAC101 0 0 0 0 06122 HAD6 0 0 0 0 06132 HAD8 0 0 0 0 06141 DUT101 0 0 0 0 06232 HPP1 0 0 0 0 06236 NGL3 0 0 0 0 06251 PPP4 0 0 0 0 06418 PTC6 -One 0 0 -One 06499 DPP101 0 0 0 0 06587 OCA2 0 0 0 0 06647 CDC1 0 0 0 0 06698 HAD9 0 0 0 0 06966 AKP1 0 0 0 0 06967 APH2 0 0 0 0 06969 HAD7 0 0 0 0 07317 INP5202 0 0 0 0 07372 OXK1 -2 0 -One 0 07434 HAD3 0 0 0 0 07547 GPM1 0 0 0 0 07629 EPP1 0 0 0 0 07692 YND1 0 0 0 0 07740 HAD2 0 0 0 0

Table 9 below shows the results of confirming the effect of gene deletion on genotoxicity, ER stress, heavy metal stress, and cell wall/membrane stress.

Genotoxic stress ER stress Heavy
metal Cell wall/membrane stress H99 lD Name HU MMS TM DTT CDS CFW CR SDS 00057 FPP1 0 0 0 0 0 0 0 0 00076 NDX6 0 0 0 0 0 0 0 0 00146 PBY101 0 0 One 0 0 0 0 0 00182 VPS29 0 0 -One -One -2 0 0 -2 00263 PTP4 0 0 0 0 0 0 0 0 00316 HAM1 0 0 0 0 0 0 0 0 00390 YMR1 0 0 0 0 -One 0 0 -One 00408 PPH3 0 -2 0 0 0 0 0 0 00427 PTC5 0 0 0 0 0 0 0 0 00498 CDC14 0 0 0 0 0 0 0 0 00500 PTC1 0 0 0 0 0 0 0 0 00554 ISC1 -One 0 0 -One -One 0 0 0 00810 HAD4 0 -2 0 0 0 0 0 0 01054 SSU72 -3 -One 0 -3 -One -One -One -2 01177 NEM1 -3 -3 0 -2 -One -2 -2 -2 01203 YVH1 -2 -3 One -One 0 0 0 -2 01254 OCA101 -One 0 0 -2 One 0 0 0 01314 PPP1 0 0 0 0 0 0 0 0 01346 HPP2 0 0 0 0 0 0 0 0 01357 SDP101 0 0 0 0 0 0 0 0 01436 SIT4 -2 -One 2 0 One -One -One One 01496 PPT1 0 0 0 0 0 0 0 0 01498 AKP2 0 0 0 0 0 0 0 0 01518 PMU1 0 0 0 0 0 0 0 0 01519 EEP1 0 0 0 0 0 0 0 0 01532 INP52 0 0 0 0 0 0 0 0 01744 HAD1 0 0 One 0 0 -One -One -3 01823 INM1 0 0 0 0 0 0 0 0 01864 NDX5 0 0 0 0 0 0 0 0 01877 GUA1 0 0 0 -One 0 0 0 -3 01900 NDX1 0 0 0 0 -One 0 0 -One 01901 NDX2 0 0 0 0 0 0 0 0 02001 INP5203 0 0 0 0 0 0 0 0 02109 INP5201 -2 -2 2 -One One -2 -2 -2 02161 ASP1 0 0 0 0 0 0 0 0 02236 PPG1 -3 -3 -3 0 0 -2 -2 -3 02375 FIG4 0 0 0 0 -One 0 0 0 02453 PTP5 0 0 0 0 -One 0 0 0 02470 PPP2 0 0 One 0 0 0 0 One 02487 PHS1 -3 0 0 -2 -One -3 -3 -2 02490 MRE11 -3 -3 -One -One -One 0 0 0 02681 APH3 0 0 0 0 0 0 0 0 02772 PMU101 0 0 0 0 0 0 0 0 02944 APH1 0 0 0 0 0 0 0 0 02986 YSA1 0 0 0 0 0 0 0 0 03014 CAX4 0 0 0 0 0 0 0 0 03052 PTC2 -One -2 0 -One -One 0 0 One 03078 NPP1 0 0 0 0 0 0 0 0 03190 CCR4 -2 0 3 -One 0 0 0 -One 03202 CAC1 -One 0 2 -One 0 0 0 One 03222 DBR1 -2 -2 2 0 -2 -One -One -3 03262 INM101 0 0 0 0 0 0 0 0 03296 SIW14 -3 -2 -One -One 0 -One -One -2 03396 NPY1 0 0 0 0 0 0 0 0 03491 NDX3 0 0 0 0 0 0 0 0 03541 PTC7 0 0 0 0 0 0 0 0 03657 LDP1 0 0 0 0 0 0 0 0 03673 PPZ1 -One -One 0 0 -2 0 -One One 03679 ACY1 0 0 0 0 0 0 0 0 03765 TPS2 0 0 -3 0 0 0 0 -3 03871 LTP1 0 0 0 0 0 0 0 0 03893 SDP102 0 0 0 0 0 0 0 One 03949 PHO13 0 0 0 0 0 0 0 0 03963 OCA1 -2 0 -One 0 0 0 0 -One 03984 YCH1 0 0 0 0 0 0 0 0 03996 GDA1 -One 0 -2 -2 -One 0 0 -2 04049 PPP5 0 0 0 0 0 0 0 0 04104 AKP3 0 0 0 0 0 0 0 0 04113 PTP3 0 0 -2 0 0 0 0 -One 04221 FBP26 -One 0 0 -One 0 -One -One -3 04224 PSR1 0 0 0 0 One 0 0 -2 04268 APN2 0 -One 0 -One 0 0 0 0 04354 XPP1 0 -One 0 0 0 -One -One -One 04475 HPP3 0 0 0 0 0 0 0 0 04718 PCD101 0 0 0 0 0 0 0 0 04731 PCD102 0 0 0 0 0 0 0 0 04732 PCD1 0 0 0 0 0 0 0 0 04782 PPU1 0 0 0 0 0 0 0 0 04796 CNA1 -3 0 -3 -3 -One -3 -3 -3 04833 TEP1 0 0 0 0 0 0 0 0 04852 NDX4 0 0 0 0 0 0 0 0 05080 CWH43 0 0 0 0 0 0 0 0 05155 PTP2 -2 -One -2 -One -2 -One -2 0 05286 LDP2 0 0 One -2 One 0 0 0 05301 CRN1 0 0 0 0 0 0 0 0 05306 PTC701 0 0 0 0 0 0 0 0 05529 HAD5 0 0 0 0 0 0 0 0 05567 PPP3 0 0 0 0 0 0 0 0 05568 PRY1 0 0 0 0 0 0 0 0 05892 SHB17 0 0 0 0 0 0 0 0 06064 PTP1 0 0 0 0 -2 0 0 One 06065 SAC101 0 0 0 0 0 0 0 0 06122 HAD6 0 0 0 0 0 0 0 0 06132 HAD8 0 0 0 0 0 0 0 0 06141 DUT101 0 0 0 0 0 0 0 0 06232 HPP1 0 0 0 0 0 0 0 0 06236 NGL3 0 0 0 0 0 0 0 0 06251 PPP4 0 0 0 0 0 0 0 0 06418 PTC6 0 0 0 0 0 0 0 -One 06499 DPP101 0 0 0 0 0 0 0 0 06587 OCA2 0 0 0 0 0 0 0 0 06647 CDC1 0 0 -One 0 0 0 0 -One 06698 HAD9 0 0 0 0 0 0 0 0 06966 AKP1 0 0 0 0 0 0 0 0 06967 APH2 0 0 0 0 0 0 0 0 06969 HAD7 0 0 0 0 0 0 0 0 07317 INP5202 0 0 0 0 2 0 0 -One 07372 OXK1 0 One 2 -One One 0 0 -One 07434 HAD3 0 0 0 0 0 0 0 0 07547 GPM1 0 0 0 0 0 0 0 0 07629 EPP1 0 0 0 0 0 0 0 0 07692 YND1 0 0 -One 0 3 0 0 -One 07740 HAD2 0 0 0 0 0 0 0 0

Table 10 below shows the results of confirming the effect of gene deletion on antifungal agent susceptibility.

Antifungal drugs susceptibility H99 lD Name AMB FCZ 5FC FDX 00057 FPP1 0 One 0 0 00076 NDX6 0 0 0 0 00146 PBY101 0 One 0 0 00182 VPS29 0 0 0 0 00263 PTP4 0 0 0 0 00316 HAM1 0 0 0 0 00390 YMR1 -One 0 0 -One 00408 PPH3 0 -One 0 0 00427 PTC5 0 0 0 0 00498 CDC14 0 0 0 0 00500 PTC1 0 -One 0 0 00554 ISC1 -2 0 0 -2 00810 HAD4 0 0 0 0 01054 SSU72 -One 0 -One -3 01177 NEM1 -One -3 -2 -3 01203 YVH1 0 0 0 -2 01254 OCA101 0 3 0 -One 01314 PPP1 0 0 0 0 01346 HPP2 0 One 0 One 01357 SDP101 0 0 0 0 01436 SIT4 -One 0 -3 -2 01496 PPT1 0 0 0 0 01498 AKP2 0 0 0 0 01518 PMU1 0 0 0 0 01519 EEP1 0 0 0 0 01532 INP52 0 0 0 0 01744 HAD1 -One 0 -One 0 01823 INM1 0 One 0 0 01864 NDX5 0 0 0 0 01877 GUA1 -3 0 -3 -3 01900 NDX1 0 0 0 0 01901 NDX2 0 0 0 0 02001 INP5203 0 0 0 0 02109 INP5201 -2 -2 -2 -2 02161 ASP1 0 0 0 0 02236 PPG1 -One -2 -3 -3 02375 FIG4 0 0 0 0 02453 PTP5 0 0 0 0 02470 PPP2 0 0 One 0 02487 PHS1 -3 -3 0 -One 02490 MRE11 0 -One -One 0 02681 APH3 0 0 0 0 02772 PMU101 0 0 0 0 02944 APH1 0 0 0 0 02986 YSA1 0 0 0 0 03014 CAX4 0 0 0 0 03052 PTC2 0 0 -2 -3 03078 NPP1 0 0 0 One 03190 CCR4 -2 -One 3 -One 03202 CAC1 0 -One 0 One 03222 DBR1 -2 -One -2 0 03262 INM101 0 0 0 0 03296 SIW14 0 -3 0 -One 03396 NPY1 0 0 0 0 03491 NDX3 0 0 0 0 03541 PTC7 0 0 0 0 03657 LDP1 0 0 0 0 03673 PPZ1 0 0 One -2 03679 ACY1 0 0 0 0 03765 TPS2 0 0 -One One 03871 LTP1 0 0 0 0 03893 SDP102 0 0 0 -One 03949 PHO13 0 0 0 0 03963 OCA1 0 -2 0 -2 03984 YCH1 0 0 0 0 03996 GDA1 -One 0 -3 -2 04049 PPP5 0 0 0 One 04104 AKP3 0 0 0 0 04113 PTP3 0 -2 0 0 04221 FBP26 -2 One -One -One 04224 PSR1 0 0 0 3 04268 APN2 0 0 -2 0 04354 XPP1 0 0 0 One 04475 HPP3 0 0 0 0 04718 PCD101 0 0 0 0 04731 PCD102 0 0 0 0 04732 PCD1 0 0 0 0 04782 PPU1 0 0 0 0 04796 CNA1 0 -2 -3 -2 04833 TEP1 0 0 0 0 04852 NDX4 0 0 0 0 05080 CWH43 0 0 0 0 05155 PTP2 0 -2 -2 -3 05286 LDP2 0 0 0 0 05301 CRN1 0 0 0 0 05306 PTC701 0 0 0 0 05529 HAD5 0 0 0 0 05567 PPP3 0 0 0 0 05568 PRY1 0 0 0 0 05892 SHB17 0 0 0 0 06064 PTP1 0 0 0 0 06065 SAC101 0 0 0 0 06122 HAD6 0 0 0 0 06132 HAD8 0 0 0 0 06141 DUT101 0 0 0 0 06232 HPP1 0 0 0 0 06236 NGL3 0 0 0 0 06251 PPP4 0 0 0 0 06418 PTC6 0 -2 0 -2 06499 DPP101 0 0 0 0 06587 OCA2 0 0 0 0 06647 CDC1 0 0 0 -One 06698 HAD9 0 0 0 0 06966 AKP1 0 0 0 0 06967 APH2 0 0 0 0 06969 HAD7 0 0 0 0 07317 INP5202 -2 -One 0 -One 07372 OXK1 0 0 0 -One 07434 HAD3 0 0 0 0 07547 GPM1 0 0 0 0 07629 EPP1 0 0 0 0 07692 YND1 0 0 -One -2 07740 HAD2 0 0 0 0

According to phenotypic analysis, 60 genetic variants (approximately 53%, 60/114) out of 114 genes showed at least one identifiable phenotype. Of the phosphatases exhibiting phenotypic changes, 43 genes (approximately 72%, 43/60) had not previously been functionally analyzed.

As a result of comparing the phosphatase development data with the corresponding BLAST matrix data, the 25 putative essential phosphatases that could not be disrupted in Example 2 and the phosphatases exhibiting multiple phenotypic traits showed evolutionary It was found that the degree of preservation was relatively higher.

In addition, each phosphorus in tissues (lung, brain, spleen and kidney) obtained 3, 7, 14 and 21 days post-infection (dpi) by wild-type C. neoformans strain H99S. By evaluating the expression levels of phatase genes, the transcriptional regulation of each of the 139 phosphatase genes was monitored during infection in a murine model. 139 new phosphatase probes and nCounter gene expression profiles were used for transcriptional pattern monitoring. The in vivo expression level of each phosphatase was normalized to the average in vivo expression level of the eight housekeeping genes and compared to the level under basal growth conditions.

During the initial infection with strain H99S (3-7 days after infection, 3-7 dpi), the in vivo expression of a large number of phosphatase genes in the lung, brain, kidney and spleen was increased, and phosphatases involved in the expression trait The in vivo expression of each gene of the enzyme also changed. In general, in the initial infection stage of the intranasal inhalation infection model, cryptococcal CFU is rarely detected in the brain and other organs except the lung due to the detection limit of the existing fungal burden assay. However, in this study, we used the Nanostring nCounter platform, which can detect single gene transcripts without amplification. Thus, if a small number of C. neoformans cells spread bloodstream from the lungs to other organs during the initial infection period, phosphatase transcripts could be detected 3 to 7 days post infection (dpi).

Example 4: Phosphatase governing the pathogenicity of C. neoformans

Virulence analysis using Galleria mellonella larval model and (2) murine based on signature-tagged mutagenesis (STM) to identify phosphatases required for the pathogenicity of C. neoformans. A large-scale infectivity and toxicity assay was performed by performing an infectivity assay on the lung and brain of an inhalation infection model.

As a result of virulence analysis on insects, 23 phosphatase genes ( CNA1, TPS2, CAC1, PTP2, VPS29, PPH3, GDA1, YVH1, SSU72, PHS1, SIW14, DBR1, PSR1, YND1, INP5201, INP5202, HAD1, SIT4, PPG1 , GUA1, NEM1, FBP26, OCA101 ) decreased mortality. Therefore, these genes were identified as related to virulence. (See Fig. 2)

As a result of STM-based mouse lung and brain infectivity assay, 24 phosphatase genes ( TPS2, CNA1, VPS29, PHS1, SIW14, CAC1, SSU72, PPG1, INP5201, CCR4, DBR1, GUA1, YMR1, NEM1, OCA1, PTP2, FBP26 , PSR1, MRE11, CDC1, XPP1, SDP102, SDP101, OCA101 ) were confirmed to be associated with infectivity because the STM score was lower than ste50 in lung and brain, of which 67% (16/24, TPS2, CNA1, VPS29, PHS1 , SIW14, CAC1, SSU72, PPG1, INP5201, DBR1, GUA1, NEM1, PTP2, FBP26, PSR1, OCA101) overlapped with genes identified in the insect model virulence assay. (See Figure 3, Ire1 is a control for lowered infectivity, and ste50 represents WT)

Among them, CNA1 (catalytic subunit of calcineurin), CAC1 (adenylyl cyclase), HAD1 (halo acid dehalogenase), and PTP2 (phosphotyrosine phosphatase) have been shown to be associated with the virulence and virulence of C. neoformans in previous studies. Since it has been reported to be related, it increases the reliability of the analysis results.

Thirty-one pathogenicity-associated phosphatase mutations ( CNA1 , TPS2 , CAC1 , PTP2 , VPS29 , PPH3 , GDA1 , YVH1 , SSU72 , PHS1 , SIW14 , DBR1 , PSR1 , YND1 , INP5201 , INP5202 ) were pathogenic in insect and murine inhalation infection models. , HAD1 , SIT4 , PPG1 , GUA1 , NEM1 , FBP26 , OCA101 , CCR4 , YMR1 , OCA1 , MRE11 , CDC1 , XPP1 , SDP102 , SDP101 ), 30 of which, excluding SDP101, showed at least one phenotypic trait, many of which showed higher expression in the lung than in other infected tissues.

The in vitro phenotype confirmation results of 31 genes are as follows. membrane integrity (26/31; 84%), DNA damage response (20/31; 65%), melanogenesis (18/31; 58%), cell wall integrity (13/31; 42%) and grown at 37 °C (14/31; 45%).

However, the sdp101 Δ mutation did not show an in vitro phenotype. Deletion of SDP102, a dual-specificity MAPK phosphatase similar to SDP101 , results in slight phenotypic alterations and reduced infectivity, suggesting that SDP101 and SDP102 may have redundant roles in the pathogenicity of C. neoformans. suggests An attempt was made to construct an spd101Δ sdp102Δ double mutant strain to confirm the functional correlation between SDP101 and SDP102 , but without success.

Likewise, INP5201 is similar to INP5202 , but inp5201 Δ inp5202 Δ double mutants could not be constructed. This may be because the inp5201 Δ mutant showed severe growth defects at 30 °C.

Therefore, mutations in SDP101 and SDP102 , and mutations in INP5201 and INP5202 may have a synthetic lethal relationship.

Conversely, OCA1 and OCA101 are also similar, but a successful oca1 Δ oca101 Δ double mutant was obtained. However, we did not find any additive or synergistic phenotypic traits in the oca1Δ oca101Δ double mutant compared to the single mutant. Therefore, OCA1 and OCA101 can independently contribute to the pathogenicity of C. neoformans.

Five of the 31 pathogenicity-associated phosphatases ( TPS2, SIW14, HAD1, OCA101, and OCA1 ) have no apparent ortholog in humans. Therefore, these five pathogenicity-related phosphatases can be excellent anti-cryptococcal targets. Since TPS2, HAD1 and OCA1 also play an important role in the virulence of C. albicans , drugs targeting TPS2, HAD1 and OCA1 may have broad antifungal activity.

Example 5: Phosphatases involved in growth at mammalian body temperature

The 31 kinds of C. neoformans pathogenicity-related phosphatases (CNA1, TPS2, CAC1, PTP2, VPS29, PPH3, GDA1, YVH1, SSU72, PHS1, SIW14, DBR1, PSR1, YND1, INP5201, INP5202, HAD1, SIT4, PPG1, GUA1, NEM1, FBP26, OCA101, CCR4, YMR1, OCA1, MRE11, CDC1, XPP1, SDP102, SDP101) were investigated.

First, since heat resistance to mammalian body temperature is an important virulence factor for human fungal pathogens, the growth of each mutant was quantitatively measured at 30 °C and 37 °C. The growth of gua1 Δ, yvh1 Δ, fbp26 Δ, siw14 Δ, dbr1 Δ, ccr4 Δ, ppg1 Δ, nem1 Δ and inp5201 Δ mutants was inhibited at both 30°C and 37°C. (See Table 5 and FIG. 4 above) The ccr4 Δ, ppg1 Δ, nem1 Δ, dbr1 Δ and inp5201 Δ mutants showed more growth defects at 37° C. than at 30° C. The growth of ssu72 Δ, phs1 Δ, mre11 Δ, tps2 Δ and cna1 Δ mutants was inhibited at 37°C but not affected at 30°C.

A total of 14 phosphatase mutants ( gua1 Δ, yvh1 Δ, phs1 Δ, nem1 Δ, cna1 Δ, ppg1 Δ , siw14 Δ, inp5201 Δ, ccr4 Δ, dbr1 Δ, ssu72 Δ, tps2 Δ, mre11 Δ, fbp26 Δ ) was reduced in growth at 37 ° C. compared to WT, and reduced murine infectivity or insect toxicity. Among them, the ppg1 Δ, cna1 Δ and tps2 Δ mutants showed the most severe growth defects at 37° C., and did not grow to the WT level even when the culture period was further extended. The ppg1 Δ, cna1 Δ and tps2 Δ mutants showed greatly reduced STM values in lung and brain. The oca101 Δ mutant showed slight growth impairment at 30 °C but not growth inhibition at 37 °C, indicating that the pathogenic role of Oca101 in C. neoformans is not related to temperature. Taking the above information together, it is suggested that the growth ability of C. neoformans at 37° C. is closely related to the pathogenicity.

Example 6: Phosphatases involved in melanin and capsule production

C. neoformans has a polyphenol pigment, melanin, and a polysaccharide capsule as virulence factors, both of which contribute to antiphagocytic activity. Among them, melanin pigment also acts as an antioxidant by scavenging activity of reactive oxygen species.

6-1. Phosphatase involved in melanin production

Thirteen of 19 phosphatase mutants defective in melanogenesis in Niger seed medium ( mre11 Δ, ccr4 Δ, vps29 Δ, yvh1 Δ, fbp26 Δ, inp5201 Δ, cac1 Δ, ptp2 Δ, ptc2 Δ, dbr1 Δ, ppg1 Δ, nem1 Δ and gua1 Δ) showed melanogenesis defects even in L-DOPA and epinephrine medium (see Table 6 and FIG. 5).

Except for the ptc2Δ mutant, infectivity or virulence were reduced in the remaining mutants, suggesting that melanogenesis is strongly correlated with pathogenicity.

In contrast, SIW14 deletion increased melanin production in all melanin-inducing media (Fig. 5) but rather attenuated virulence (Fig. 2), suggesting that other cellular functions mediated by SIW14 may promote virulence.

Next, we investigated whether these phosphatases were directly involved in inducing expression of the LAC1 gene encoding laccase, the rate-limiting enzyme for melanin production in C. neoformans under nutrient deficiency conditions.

According to FIG. 6, deletion of PTP2 , CCR4 , INP5201 , CAC1 , DBR1 , FBP26 , GUA1 , NEM1 and PPG1 markedly reduced the degree of induction of LAC1 gene expression during nutrient deficiency, and in particular, deletion of PTP2 , CCR4 , CAC1 and PPG1 strongly suppressed the induction of LAC1 gene expression.

Ptp2 has been reported as a negative feedback regulator of Hog1 MAPK. Hog1 deletion increases LAC1 induction and melanogenesis. Recently, it has been reported by the present inventors that the induction of expression of the LAC1 gene is controlled by four key TFs: Bzp4, Usv101, Hob1 and Mbs1. Among them, the expression of BZP4 and HOB1 genes is induced by nutrient deprivation, and BZP4 induction is dependent on HOB1 . Therefore, we investigated whether phosphatase, which regulates the degree of induction of LAC1 gene expression, regulates the induction of BZP4 and HOB1 genes. In most of the melanin-deficient phosphatase mutants, the degree of induction of BZP4 gene expression by nutritional deprivation was markedly reduced, but in particular, CCR4 deletion greatly reduced the degree of induction of BZP4 expression, and in particular, greatly suppressed the degree of induction of HOB1 expression by nutritional deprivation. did (FIG. 7). Thus, the Ccr4-Hob1-Bzp4-dependent signaling pathway appears to play an important role in the induction of LAC1 gene expression and melanogenesis in C. neoformans .

6-2. Phosphatase involved in capsule production

Phosphatases involved in capsule production were investigated. Capsule production was measured quantitatively by packed cell volume. Deletion of Cac1, Ptp2, Psr1, Ndx5, Hpp2, Oca1, Sdp102, Ynd1, Ngl3, Ppp5 and Cwh43 reduced the concentrating cell volume and Inp5201, Dbr1, Phs1, Cdc1, Cna1, Sit4, Ccr4, Nem1, Pcd102 and Fig.4 Deletion of increased the concentrated cell volume. (See Table 6 and Figure 8)

The actual capsule thickness of these mutants was investigated. Of these, 6 mutants ( cac1 Δ, sdp102 Δ, ndx5 Δ, ptp2 Δ, ynd1 Δ and psr1 Δ) exhibited reduced capsule production consistent with a phenotypic decrease in condensate cell volume in cell volume analysis, of which 5 Dogs ( cac1 Δ, ptp2 Δ, sdp102 Δ, ynd1 Δ, psr1 Δ) showed reduced virulence. In contrast, the inp5201 Δ mutant had greatly enhanced capsule production but greatly reduced virulence (STM < -7), which is thought to be due to a defect in melanin production.

Overall, the ability to produce melanin and polysaccharide capsules showed a high correlation with the pathogenic potential of C. neoformans.

Example 7: Virulence of retromer complexes and C. neoformans

Among virulence-related phosphatases, Vps29 (CNAG_00182) is presumed to be a component of a retromer complex first discovered in S. cerevisiae .

Yeast retromers are cytosolic heteropentameric protein complexes that mediate the intracellular transport of proteins from post-Golgi organelles to the lytic compartment. Yeast retromers are composed of a Cargo - Recognition Core (CRC) complex containing Vps29, Vps35 and Vps26 and a Membrane-Deforming Sorting Nexin (SNX) complex containing Vps5 and Vps17.

To determine whether the function of Vps29 resulted from a conserved role in the retromeric complex, we functionally characterized the other retromeric components of C. neoformans. All remaining retromeric component genes encoding proteins homologous to Vps35 (CNAG_01837), Vps26 (CNAG_01426), Vps5 (CNAG_01315) and Vps17 (CNAG_00508) were identified in the C. neoformans genome. This suggests that the retromeric complex is evolutionarily conserved in pathogenic fungi.

Each gene was deleted from the H99 strain and phenotypic analysis was performed. Deletion of VPS35 and VPS26 resulted in more dramatic phenotypic changes than deletion of VPS29 , indicating that Vps35 and Vps26 are important CRC complex components in C. neoformans. In contrast, SNX components VPS5 and VPS17 deletions showed relatively little phenotypic change compared to VPS29 deletion (FIG. 9).

In the insect killing assay, the virulence of the CRC complex mutant was significantly reduced, whereas the virulence of the SNX complex mutant was not significantly different from that of the wild-type strain. (FIG. 10) However, in the murine-based STM assay, the lung and brain infectivity of CRC complex mutants and SNX complex mutants was significantly reduced. (FIG. 11) Taken together, the experimental results indicate that both the retromeric CRC complex and the SNX complex are important for the pathogenicity of C. neoformans.

Example 8: C. neoformans O- gda1 and ynd1 regulate mannosylation

In S. cerevisiae , Gda1 and Ynd1 affect the antiport exchange ratio between GDP-mannose and GMP, resulting in O- linked and N -linked glycoproteins and glycosphingolipids. It is a golgi membrane bound apyrase that regulates mannosylation. Gda1 shows a highly specific activity for GDP, while Ynd1 shows a much broader spectrum of activity. Therefore, double deletion of GDA1 and YND1 results in more severe glycosylation defect than single deletion.

To investigate whether the gda1Δ and ynd1Δ mutants of C. neoformans exhibit defects in O- glycosylation, we compared the O- glycan profiles of wild-type, gda1Δ and ynd1Δ strains.

O- linked oligosaccharides bound to cwMPs, which are cell wall mannoproteins of C. neoformans, were obtained by hydrazinolysis and prepared with 2-aminobenzoic acid (2-AA). Labeled and analyzed using HPLC with fluorescence detection.

According to FIG. 12, the O- linked oligosaccharide of C. neoformans wild-type strain was mainly composed of 2 to 4 mannose residues (Man2-Man4; M2-M4), and some xylose (X1M2-X1M4) was included . - O- glycan species and some α1,2-mannotriose (α1,2-mannotriose, M3*) species were included. However, the O- glycan profile of the gda1Δ mutant showed a dramatic decrease in the M4 peak and an increase in the M1 peak compared to the wild-type strain. In particular, xylose-containing species, which had a low content in the wild type, increased in O- glycan of the gda1 Δ strain.

According to FIG. 13, the ynd1 Δ mutant showed an increase in the M1 peak and a slight decrease in the M4 peak, but the reduction did not lead to a decrease in the M4 peak in the gda1 Δ mutant. The change in the O- glycan profile is serious in the elongation process of O- glycan as the supply of GDP-mannose from the cytoplasm to the lumen of the Golgi is reduced due to the decrease in GDPase activity in the gda1 Δ strain and the ynd1 Δ strain. indicates that there is a defect. (See FIG. 14) Based on the above experimental results, gda1 and ynd1 of C. neoformans are thought to encode membrane bound apyrase required for Golgi N-glycosylation and O- glycosylation.

gda1Δ had a dramatic decrease in major O- glycans more than ynd1Δ , implying that the Gda1 protein is a major GDPase responsible for supplying GDP-mannose in the major O- glycan biosynthetic pathway. The reduced capsule size of ynd1 Δ suggests that Ynd1 protein may also be involved in supplying GDP-mannose for capsule biosynthesis. However, the gda1 Δ ynd1 Δ double mutant was not obtained. This means that the two proteins are synthetically lethal in C. neoformans. In the insect killing assay, both the gda1 Δ and ynd1 Δ mutants showed reduced virulence, suggesting that O -mannosylation is important for the virulence of C. neoformans.

Example 9: Phosphatases involved in blood-brain barrier (BBB) crossing

The biggest problem with human infection of C. neoformans is brain infection that causes fatal meningoencephalitis. Brain STM scores for all phosphatase mutants were generally similar to lung STM scores, but some showed lower brain STM scores. (See Figure 3: TPS2 , YMR1 , FBP26 , MRE11 , CDC1 and XPP1 ). However, since phosphatase mutant strains administered through the intranasal inhalation pass pass through the lungs first, the mutant strains that reach the brain are not uniformly distributed compared to the inhaled mutant pool, so the lung and brain STM scores are directly calculated. There were problems with which it was difficult to compare.

In order to investigate the role of phosphatase in brain infection, growth at 37 ° C. was used to cross the BBB (traverse ) ability was monitored.

According to FIG. 15, five mutants ( xpp1 Δ, ssu72 Δ, siw14 Δ, sit4 Δ and gda1 Δ) showed significantly reduced ability to cross the BBB. Since adhesion to the BBB may be a prerequisite for effective BBB passage, a BBB-adhesion assay was performed. According to FIG. 16, xpp1 Δ, ssu72 Δ, siw14 Δ, and sit4 Δ mutants, except for gda1 Δ, showed reduced BBB attachment. According to the experimental results, it is thought that reduction of O- mannosylation of proteins or lipids by gda1 Δ affects BBB passage but not BBB attachment.

When C. neoformans was cultured at 37°C in a tissue culture medium used for in vitro BBB passage and adhesion analysis, it was found that growth was not active. This may be due to the low glucose concentration (0.1% glucose).

In a recent paper, the present inventors identified a TF ( PDR802 ) that can promote BBB attachment by in vitro host-mimic conditions (HMC (host-mimic conditions); 5% CO ₂ , 37° C., RPMI medium containing 10% FBS). , FZC31 and GAT201 ) were reported to be able to induce expression.

Based on this, expression of brain infection-related genes (inositol transporter genes ( ITR1a and ITR3c ) and MPR1 ) and the TF gene were induced under host mimicry conditions (HMC) in xpp1 Δ, ssu72 Δ, siw14 Δ and sit4 Δ mutants Investigated if it could be.

According to FIG. 17, deletion of SSU72 most significantly reduced HMC-mediated induction of MPR1 expression. This suggests that the defect in BBB passage and attachment of the ssu72Δ mutant may be caused by decreased induction of MPR1 expression. However, according to FIG. 16, since the ssu72 Δ mutant has more defects in BBB passage and attachment than the mpr1 Δ mutant, other cellular functions of Ssu72 may be involved in BBB passage and attachment. SIT4 deletion and GDA1 deletion markedly reduced HMC-mediated induction of ITR3c expression. GDA1 deletion also affected FZC31 induction. However, the expression of GAT201 and PDR802 was not affected by deletion of SIT4 , SIW14 , SSU72 , GDA1 or XPP1 .

To comprehensively understand the signaling network associated with brain infection by C. neoformans, combining research data on phosphatases, TFs, and kinases involved in BBB passage and using STRING analysis to generate functional gene networks did.

According to FIG. 18, genes related to glucose sensing (Sit4-Snf1-Gal83), RNA processing (Ssu72), and purine metabolism (Xpp1-Met3) were found to be important for C. neoformans to pass through the BBB. did.

Taken together, the above experimental results indicate that C. neoformans utilizes a complex signaling network for brain infection.

Example 10: Comparison of phosphatases related to fungal pathogenicity

The virulence data of C. neoformans phosphatase mutants and data of other plant or animal pathogenic fungi were compared to select key pathogenicity-related phosphatases.

According to FIG. 19, 13 phosphatases (Cna1/Cmp1, Sit4, Oca1, Yvh1, Sdp101/Cpp1, Ptp2/Ptp3, Cac1) matched in human pathogens Cryptococcus neoformans and Candida albicans. /Cyr1, Ccr4, Had1/Rhr2, Tps2, Inp5201/Inp51, Ppg1 and Gua1) are thought to be key phosphatases for pathogenicity. Among the 13 phosphatases, CnaA, SitA and OrlA (Tps2 ortholog) were confirmed to be involved in the virulence of Aspergillus fumigatus . (SitA is the ortholog of Sit4 in A. fumigatus)

Eight phosphatases (Sit4, Yvh1, Sdp2 / Msg5 (Sdp101 orthologue), Ptp2, Ac1 (Cac1 orthologue), Tps2, Inp53 (Inp5201 orthologue) and Ppg1) were found to be required. Sit4 is involved in the TOR pathway, Cac1 in the cAMP pathway, and Ppg1 and Yvh1 involved in cell growth, nutrient sensing and stress response in fungal pathogens. The Cna1 and Had1-mediated calcineurin pathway, the Tps2-mediated trehalose pathway, and the Msg5-mediated Mpk1/Slt2 MAPK pathway are all required to maintain cell wall integrity. Ptp2, a major negative feedback regulator of the HOG pathway, is involved in the adaptation and stress response of fungal pathogens. Inp51, Inp52 and Inp53 are involved in phosphoinositide signaling that controls vesicle trafficking, actin cytoskeleton and cell wall integrity.

Based on these data, phosphatases and signaling pathways involved in cell growth, nutrient sensing, cell wall integrity, stress response and phosphoinositide signaling are believed to play pivotal roles in fungal pathogenicity.

However, PPH3 deletion reduces the virulence of C. neoformans and F. graminearum but enhances the virulence of C. albicans . In contrast, deletion of ptc2 or ptc3 reduces the virulence of C. albicans or F. graminearum, respectively, but not C. neoformans . Therefore, phosphatase may have a difference in the degree of involvement in virulence for each type of fungus.

Fifty-four mutants that did not show phenotypic changes did not show infectivity and virulence defects. These 54 phosphatases are not considered to be involved in the pathological function of C. neoformans.

Example 11: Identification of phosphatases related to cell membrane stability

To analyze the cell membrane stress susceptibility of each phosphatase mutant to stress conditions in vitro, C. neoformans was grown at 30 °C for 16 h, serially diluted 10-fold (1-10 ⁴ ), and cell membrane Spot inoculation was performed on YPD agar medium containing a stress-inducing chemical (SDS; 0.03% - 0.05%). Cells were cultured at 30° C. for 1 to 5 days and photographed daily.

20 and 21, VPS29, YMR1, SSU72, NEM1, YVH1, SIT4, GUA1, INP5201, PHS1, DBR1, SIW14, SDP102, OCA1, GDA1, FBP26, PSR1, CDC1, INP5202, or YND1 genes are deleted. Compared to WT, C. neoformans showed significantly reduced growth in a medium supplemented with SDS (sodium dodecyl sulfate), a cell membrane stress inducer. When other phosphatase genes were deleted, their growth was not significantly different from that of WT. (data not disclosed) Therefore, the VPS29, YMR1, SSU72, NEM1, YVH1, SIT4, GUA1, INP5201, PHS1, DBR1, SIW14, SDP102, OCA1, GDA1, FBP26, PSR1, CDC1, INP5202, and YND1 genes It can be seen that it is a phosphatase related to resistance to stress and cell membrane stability.

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Phosphatase that modulates cell membrane stability of Cryptococcus neoformans, and method for screening antifungal agents using the same <130> PN200402-P6 <150> KR 10-2020-0055355 <151> 2020-05-08 <160> 238 <170> KoPatentIn 3.0 <210> 1 <211> 204 <212> PRT <213> artificial sequence <220> <223> vps29 >CNAG_00182 Transcript 1 <400> 1 Met Val Leu Val Leu Val Ile Gly Asp Leu His Ile Pro Asn Leu Val 1 5 10 15 His Asp Leu Pro Ala Lys Phe Lys Lys Leu Leu Val Pro Gly Lys Ile 20 25 30 Gly Gln Ile Ile Cys Thr Gly Asn Val Cys Asp Lys Glu Thr Tyr Asp 35 40 45 Tyr Leu Arg Thr Thr Ala Pro Glu Val His Val Val Arg Gly Glu Phe 50 55 60 Asp Glu Asn Pro His Phe Pro Leu Ser Leu Ile Ile Gln His Gln Ser 65 70 75 80 Leu Arg Ile Gly Val Val His Gly Gln Gln Val Val Pro Ala Gly Asp 85 90 95 Pro Asp Met Leu Ala Ala Leu Ala Arg Gln Met Asp Val Asp Val Leu 100 105 110 Ile Ser Gly Gly Thr His Arg Phe Glu Ser Phe Glu Phe Glu Gly Arg 115 120 125 Phe Phe Val Asn Pro Gly Ser Ala Thr Gly Ala Trp Ser Ser Leu Trp 130 135 140 Asn Gly Glu Val Thr Pro Ser Phe Ala Leu Met Asp Ile Gln Gly Pro 145 150 155 160 Val Ile Val Thr Tyr Val Tyr Gln Leu Val Asp Gly Glu Val Lys Val 165 170 175 Asp Lys Val Glu Tyr Arg Lys Pro Asp Leu Thr Ser Glu Thr Gln Ser 180 185 190 Gln Ser Thr Arg Ser Glu Val Ala Ala Arg Trp *** 195 200 <210> 2 <211> 785 <212> PRT <213> artificial sequence <220> <223> YMR1 >CNAG_00390 Transcript 1 <400> 2 Met Asp Ala Leu Arg Val Ala Arg Val Asp Asn Val Thr Ile Gln Tyr 1 5 10 15 Phe Leu Pro Pro Thr Ala Pro Asp Gln Lys Pro Thr Pro Leu Thr Gln 20 25 30 Ile Gly Gln Leu His Leu Thr Pro His Leu Ile Phe Ser His Thr 35 40 45 Pro Ser Thr Ala Tyr Glu Pro Glu Ile Trp Ile Pro Tyr Pro Leu Ile 50 55 60 Thr Arg Leu Thr Arg Leu Pro Gln Thr Ile Asn Gly Leu Tyr Pro Leu 65 70 75 80 Gln Val Glu Thr Lys Thr Phe Glu Ser Tyr Val Leu Leu Phe Thr Lys 85 90 95 Asp Arg Asp Asp Gly Ala Glu Glu Val Trp Gln Ser Val Lys Asp Cys 100 105 110 Ser Val Lys Ser Ser Val Glu Gln Leu Tyr Ala Phe Phe Tyr Val Pro 115 120 125 Pro Ser Pro Gly Thr Gly Trp Thr Val Phe Asn His Arg Thr Glu Phe 130 135 140 Ala Arg Gln Gly Leu Gly Thr Arg Thr Lys Ala Trp Arg Phe Thr Asp 145 150 155 160 Ile Asn Lys Asp Tyr Ser Phe Ser Pro Thr Tyr Pro Ser Lys Leu Val 165 170 175 Val Pro Ser Arg Ile Ser Asp Ser Thr Leu Met Tyr Ala Gly Lys Tyr 180 185 190 Arg Ser Lys Ala Arg Ile Pro Ala Leu Thr Tyr Leu His Trp Ala Asn 195 200 205 Asn Ala Ser Ile Thr Arg Ser Ser Gln Pro Met Val Gly Ile Lys Asn 210 215 220 Ser Arg Ser Ser Gln Asp Glu Arg Leu Val Glu Cys Ile Phe Ser Ser 225 230 235 240 His Met Phe Leu Asp Asn Ala Tyr Ser Ser Ala Pro Ile Phe Gly Ala 245 250 255 Thr Ser Thr Asn Leu Ile Ile Asp Ala Arg Pro Thr Thr Asn Ala Met 260 265 270 Ala Asn Val Ala Met Gly Ala Gly Thr Glu Asn Met Glu Asn Tyr Lys 275 280 285 Leu Gly Lys Lys Ala Tyr Leu Gly Ile Asp Asn Ile His Val Met Arg 290 295 300 Asn Ser Leu Lys Thr Val Ala Glu Ala Ile Arg Glu Ala Asn Leu Arg 305 310 315 320 Pro Ser Val Pro Leu Asn Arg Ala Leu Leu Arg Lys Ser Asn Trp Leu 325 330 335 Arg His Ile Ser Thr Ile Leu Asp Gly Ala Leu Ile Ile Val Arg Asn 340 345 350 Ile His Leu Asn Ala Ser His Val Leu Ile His Cys Ser Asp Gly Trp 355 360 365 Asp Arg Thr Gly Gln Leu Ser Ala Val Ala Gln Ile Cys Leu Asp Pro 370 375 380 Tyr Tyr Arg Thr Phe Asp Gly Phe Lys Val Leu Val Glu Lys Asp Trp 385 390 395 400 Leu Ala Phe Gly His Lys Phe Leu Asp Arg Ser Gly His Leu Ser Ser 405 410 415 Glu Lys Tyr Phe Met Val Thr Glu Asn Asp Asp Asp Met Glu Glu Glu 420 425 430 Gly Val Ser Ala Gln Arg Ala Ala Gln Ala Phe Phe Ala Thr Val Gln 435 440 445 Lys Gln Phe Thr Ser Thr Ser His Leu Lys Glu Ile Ser Pro Val Phe 450 455 460 His Gln Phe Leu Asp Cys Val Arg Gln Ile Gln Arg Gln Phe Pro Glu 465 470 475 480 Arg Phe Glu Phe Asn Glu Gln Tyr Leu Leu Asp Ile Tyr Arg His Leu 485 490 495 Tyr Thr Cys Gln Phe Gly Thr Phe Leu Phe Asn Asn Glu Arg Glu Arg 500 505 510 Gln Glu Ser Ala Ser Pro Ser Arg Lys Ser Phe Val Glu Gln Thr Cys 515 520 525 Ser Val Trp Asp Tyr Leu Asp Ser Pro Ser Glu Arg Glu Lys Tyr Ile 530 535 540 Asn Ser Leu Tyr Asp Thr Thr Leu Asp Ser Asn Gln Ser Arg Asp Ala 545 550 555 560 Gly Ala Asp Gln Gly Val Leu Phe Tyr Asn Pro Lys Asp Val Arg Phe 565 570 575 Trp Phe Arg Leu Phe Gly Arg Gly Asp Glu Glu Met Asn Gly Ser Ser 580 585 590 Leu Thr Leu Asn Gln Pro Gln Gly Val Asp Ile Ile Gly Pro Ile Gly 595 600 605 Gly Asp Gln Val Glu Asp Met Ala Ala Gly Glu Ile Leu Arg Gly Ala 610 615 620 Ser Pro Val Ser Ala Pro Ser Pro His Ala Thr Ala Ser Gln Ser Arg 625 630 635 640 Ser Trp Asn Trp Ser Gln Leu Ser Gly Asn Ala Leu Asn Ala Val His 645 650 655 Ser Ala Ala Arg Glu Ile Lys Ser Ile Ser Gln Asp Ala Leu Ser Gln 660 665 670 Ile Arg Ala Glu Ala Asn Glu Leu Asp Arg Glu Ser Trp Glu Gln Asp 675 680 685 Gly Lys Gly Lys Asn Ser Glu Pro Ala Ser Leu Thr Glu Ser Thr Leu 690 695 700 Leu Pro Glu Thr Asn Pro Trp Ser Ala Glu Ala Arg Ser Ser Pro Thr 705 710 715 720 Ile Pro Pro Pro Arg Pro Asn Thr Gln Val Ser Arg Thr Thr Gln Asn 725 730 735 Pro Trp Ala Ala Met Pro Asp Thr Ile Thr Ser Leu Ser Asn Leu Thr 740 745 750 Leu Asp Gly Lys Ala Pro Gly Ser Pro Ala Asn Asp Ala Gly Thr Lys 755 760 765 Glu Arg Ala Gly Glu Lys Gln Gln Lys Ala Trp Asp Pro Leu Gly Ala 770 775 780 Leu 785 <210> 3 <211> 376 <212> PRT <213> artificial sequence <220> <223> SSU72 >CNAG_01054 Transcript 1 <400> 3 Met Asp Pro Arg Arg Arg His Asn Gln Arg Pro Pro Pro Pro Ser Ser 1 5 10 15 Ser Leu Pro Pro Asn Pro Ala Ala Tyr Asn Ala Pro Pro Asn Ser Tyr 20 25 30 Gly Gly Ser Tyr Pro Asp Ala Arg Gln Tyr Gln Gly His Asn Gly Ala 35 40 45 His Ser Thr Pro Gln Gly Tyr Arg Ser Ala Pro Pro Pro Gln Pro Pro 50 55 60 Tyr Gly Ala Leu Pro Gly Glu Gln Arg Ala Phe Pro Pro Ser Asn Met 65 70 75 80 Pro Asn Tyr Pro Pro Ser Gly Pro Pro Asp Pro Arg Met Arg Pro Ser 85 90 95 Gln Asp Pro Arg Ser Arg Leu Ser Gly Ser Gln Gly Asn Tyr Asn Thr 100 105 110 Pro Thr Pro Pro Ser Gly His Thr Pro Pro Ser Leu Pro Asn Tyr Gly 115 120 125 Thr Pro Pro Ile Ser Ala Pro Thr Ile Pro Leu Pro Ser Gln Gln Ser 130 135 140 His Gln Gln Phe Tyr Thr Pro Pro Ser Gly Pro Thr Ser Ser Leu Pro 145 150 155 160 Gly Ala Met Pro Ser Gly Val Ile Ser Glu Pro Ala Asn Gly Phe Val 165 170 175 Asp Lys Asp Val Pro Gln Gly Arg Arg Arg Pro Leu Phe Cys Val Val 180 185 190 Cys Ala Ser Asn Asn Asn Arg Ser Met Glu Ala His Tyr Val Leu Asn 195 200 205 Lys Asn Ser Phe Arg Val Val Ser Ala Gly Thr Gly Ser Ala Val Arg 210 215 220 Leu Pro Gly Pro Ala Ile Asp Lys Pro Asn Val Tyr Arg Phe Gly Thr 225 230 235 240 Pro Tyr Asp Asp Ile Tyr Arg Asp Leu Glu Ser Gln Asp Pro Gln Leu 245 250 255 Tyr Thr Arg Asn Gly Ile Leu Pro Met Leu Asp Arg Asn Arg Lys Val 260 265 270 Lys Lys Ala Pro Glu Lys Trp Gln Glu Leu Lys Ser Val Leu Ala Asp 275 280 285 Val Val Ile Thr Cys Glu Glu Arg Cys Tyr Asp Ala Val Cys Asp Asp 290 295 300 Leu Leu Thr Arg Ser Gly Glu Tyr Asn Arg Pro Ile His Ile Ile Asn 305 310 315 320 Ile Glu Ile Lys Asp Asn Pro Glu Glu Ala His Ile Ala Gly Gln Ser 325 330 335 Ile Leu Glu Leu Ala Arg Ala Ile Glu Ala Ser Asp Asp Leu Asp Ser 340 345 350 Asp Ile Asp Ala Ile Leu Asn Ala His Gly Asp Lys His Pro His Thr 355 360 365 Leu Leu His Thr Val Gly Phe Tyr 370 375 <210> 4 <211> 500 <212> PRT <213> artificial sequence <220> <223> NEM1 >CNAG_01177 Transcript 1 <400> 4 Met Asn Thr Leu Ser Arg Ile Asp Ser Tyr Phe Ser Ala Ile Ala Ser 1 5 10 15 Arg Pro Thr Thr His Pro Pro Arg Thr Pro Pro Arg Arg Ser Arg Gln 20 25 30 Thr Ile Ser Ser Ile Ser Val Pro Pro Pro Thr Ala Pro Leu Ile Leu 35 40 45 Arg Ile Ala Leu Val Leu Trp Ser Val Leu Leu Thr Val Trp Arg Ser 50 55 60 Phe Val Gly Glu Thr Arg Ala Thr Arg Arg Arg Gly Arg Arg Ser Arg 65 70 75 80 Arg Lys Arg Leu Ala Gly Leu Arg Glu Leu Gly Glu Arg Val Met Ile 85 90 95 Thr Ala Gly Ile Ala Ser Leu Asp Thr Pro Gln Glu His Thr Glu Gly 100 105 110 Asp Glu Gly Ser Glu Asp Asp Lys Glu Asp Gly Trp Val Asp Pro Val 115 120 125 Thr Arg Gly Pro Glu Gly Ser Ala Ser Leu Glu Glu Ala Pro Pro Gly 130 135 140 Glu Asp Glu Phe Val Ser Ala Asn Thr Ala Ser Thr Gly Thr Gly Ala 145 150 155 160 Ala Glu Val Glu Glu Glu Pro Glu Pro Asp Pro Asp Glu Met Thr Val 165 170 175 Thr Ala Lys Asp Asp Arg Leu Gly Gly Pro Asp Pro Asn Phe Thr Phe 180 185 190 Arg Leu Arg Ser Ala Pro Lys Lys Glu Leu Asp Gly Thr Glu Thr Ala 195 200 205 Val His Ser Pro Gly His Lys Pro Ile Pro Ser Phe Gln Arg Pro Pro 210 215 220 Ser Pro Thr Ser Ile Leu Asn Asn Pro Ile Thr Pro Ser Pro Pro Pro 225 230 235 240 Pro Pro Pro Ser Lys Thr Val Glu Pro Ser Pro Lys Arg Pro Ser Gly 245 250 255 Thr Arg Leu Leu Ala Asn Pro Ile Ser Thr Ser Leu Leu Asp Pro Ser 260 265 270 Val Pro Ala Pro Ala Ser Asn Ala Asp Ser Ser Leu Phe Arg Lys Pro 275 280 285 Ser Pro Arg Pro Leu Arg Gln Pro Thr Thr Pro Phe His Leu Gln Lys 290 295 300 Thr Leu Ile Leu Asp Leu Asp Glu Thr Leu Ile His Ser Thr Ser Arg 305 310 315 320 Pro Ile His Tyr Pro Gly Gly Ser Ser Gly Gly Gly Gly Leu Leu Gly 325 330 335 Leu Ser Val Gly Gly Val Phe Gly Asn Gly Arg Ala Lys Glu Gly His 340 345 350 Thr Val Glu Val Val Val Asn Gly Arg Ser Thr Met Tyr His Val Tyr 355 360 365 Lys Arg Pro Tyr Val Asp His Phe Leu Lys Lys Val Ala Ser Trp Tyr 370 375 380 Thr Leu Val Ile Phe Thr Ala Ser Met Pro Glu Tyr Ala Asp Pro Val 385 390 395 400 Ile Asp Trp Leu Asp Gly Gly Arg Asn Leu Phe Ala Lys Lys Leu Tyr 405 410 415 Arg Glu Asn Cys His Val Gln Pro Asn Gly Ser Tyr Ile Lys Asp Leu 420 425 430 Thr Leu Val Glu Lys Asp Leu Ser Arg Val Cys Phe Met Asp Asn Ser 435 440 445 Pro Val Ser Tyr Ser Trp Asn Lys Ala Asn Ala Leu Pro Ile Glu Gly 450 455 460 Trp Thr Ser Asp Pro Asn Asp Glu Ala Leu Leu His Ser Ile Pro Val 465 470 475 480 Leu Asp Ser Leu Arg Phe Val Asn Asp Val Arg Arg Val Leu Gly Ile 485 490 495 Arg Gly Phe Ser 500 <210> 5 <211> 711 <212> PRT <213> artificial sequence <220> <223> YVH1 >CNAG_01203 Transcript 1 <400> 5 Met Pro Gln His Asp Thr Val Pro Ser Arg Pro Pro Ser Asp Ala Ala 1 5 10 15 Lys Ser Asp Asn Leu Ala Ile Gln Asp Leu Ser Arg Glu Val Thr Ser 20 25 30 Thr Gly Asn Ile Arg Leu Ala Ala Lys Arg Leu Ala Gln Ser Ala Gln 35 40 45 Ala Glu Lys Val Arg Asn Leu Lys Glu Arg Ala Ser Thr Arg Ala Gln 50 55 60 Val Ala Gln Pro Tyr Ala Arg Trp Ala Asp Asp Pro Glu Glu Ala Glu 65 70 75 80 Tyr Leu Gln Ser Asn Val His Ala Gln Ala Ala Ala His Val Glu Asp 85 90 95 Gln Val Leu Val Ser Asp Glu Glu Glu Glu Glu Glu Glu Glu Lys Glu Ala 100 105 110 Met Gly His Met Gln Glu Val Val Asp Gly Leu Trp Val Gly Asp Leu 115 120 125 Val Ala Ala Asn Asp Asp Asp Glu Leu Glu Lys Asn Gly Ile Lys Asn 130 135 140 Ile Leu Ser Ala Leu Arg Pro Ser Leu Lys Phe Ser Asp Lys Tyr Ala 145 150 155 160 Val Tyr Pro Leu Glu Ile Asp Asp Ser Ala Asp Thr Asp Leu Leu Ser 165 170 175 His Leu Pro Ser Cys Val Ala Trp Ile Lys Glu Ile Leu Asp Leu Arg 180 185 190 Gln Lys Ala Ala Glu Pro Ser Ser Gln Lys Asn Gly Thr Glu Asn Gly 195 200 205 Glu Ser Leu Lys Arg Ser Pro Asp Ile Asp Thr Val Ala Gln Pro Gly 210 215 220 Lys Pro Gly Gly Val Leu Val His Cys Gln Ala Gly Met Ser Arg Ser 225 230 235 240 Ala Ser Ile Val Ala Ala Tyr Leu Met Ser Gln Tyr Asp Leu Asp Pro 245 250 255 Met Glu Ala Met Thr Met Ile Arg Glu Lys Arg Pro Val Val Glu Pro 260 265 270 Ser Ala Thr Phe Trp His Gln Leu Gly Leu Phe Tyr Thr Thr Asp Gly 275 280 285 Lys Val Ser Leu Lys Asp Arg Ser Thr Arg Gln Tyr Tyr Met Glu Arg 290 295 300 Thr Thr Thr Gln Phe Ile Asn Gly Asp Gly Thr Ala Pro Ser Met Glu 305 310 315 320 Lys Met Ala Lys Tyr Pro Ala Ser Pro Ser Pro Ser Asn Pro Pro Thr 325 330 335 Pro Lys Asp His Ala Arg Arg Lys Ile Arg Cys Lys Met Cys Arg Arg 340 345 350 His Leu Ala Val Arg Glu His Met Met Asp His Ile Leu Asp Gln Ala 355 360 365 Pro Pro Val Pro Ala Ser Arg Pro Arg Thr Pro Ser Gly Ala Ser Ile 370 375 380 Ser Ser Gln Arg Ala Ser Phe Ser Ser Asn Ala Gly Met Arg Phe Thr 385 390 395 400 Asp Val Val Gly Glu Gly Ala Gly Phe Leu Thr Glu Arg Glu Arg Arg 405 410 415 Gly Ser Gln Val Ser Asp Val Ile Asn Pro Leu Thr Gly Leu Pro Gly 420 425 430 Ala Leu Ser Arg Arg Ser Ser Ala Gly Ala Gly Ser Asn Gly Ala Val 435 440 445 Ser Pro Thr Ala Thr Gln Thr Leu Tyr Glu Arg Asp Thr Val Thr Ser 450 455 460 Pro Leu Ser Ile Ser His Asn His His Asn Asn Asn Asn Asn Asn Asn 465 470 475 480 Thr Thr His Pro Ala Ser Arg Arg Gly Pro Ile Leu Arg Asn His Ser 485 490 495 Glu Pro Ala Gly Thr Val Pro Pro Pro Pro Val Pro Leu Pro Ala Ala 500 505 510 His Ser Thr Thr Ser Val Pro Ala Pro Gln Ala Pro Thr Thr Gln Arg 515 520 525 Ala Leu Gln Ser Ala Asp Gln Leu Asn Met Arg Leu Pro Pro Gln Leu 530 535 540 Leu Ala Leu Arg Met Ala Gly Met Gly Gly Ala Ala Ala Asn Ala Gly 545 550 555 560 Ala Ser Ala Asn Ala Ser Asn Pro Pro Val Ser Pro Gly Thr Asn Thr 565 570 575 Pro Ser Pro Val Ile Glu Lys Glu Arg Arg Asp Gln Ser Ser Ser Ser 580 585 590 Ile Asn Thr Asn Gly Gly Ala Gly Ala Ala Ala Arg Arg Phe Ser Ser 595 600 605 Leu Ala Met Thr Pro Lys Asp Glu Lys Glu Glu Thr Lys Leu Tyr Glu 610 615 620 Arg Arg Ala Ser Gly Gly Glu Gly Met Tyr Gly Pro Pro Pro Ile Leu 625 630 635 640 Val Asn Asn Lys Cys Ser Gly Tyr Phe Val Glu Pro Leu Thr Trp Met 645 650 655 Glu Pro Val Leu Ser Lys Gly Gln Ile Ala Gly Lys Leu Val Cys Pro 660 665 670 Asn Glu Lys Cys Gly Val Lys Ile Gly Asn Phe Asp Trp Ala Gly Val 675 680 685 Gln Cys Gly Cys Lys Glu Trp Val Thr Pro Gly Phe Cys Ile His Arg 690 695 700 Ser Lys Val Asp Glu Val Phe 705 710 <210> 6 <211> 212 <212> PRT <213> artificial sequence <220> <223> OCA101 >CNAG_01254 Transcript 1 <400> 6 Met Ala Asn Lys Pro Pro Ser Gln Pro Leu Ile Gln Val Pro Ala Leu 1 5 10 15 Phe Ser Ile Val Glu Pro Gly Val Tyr Arg Ser Ala Ser Pro Thr Pro 20 25 30 Ser Gln Val Pro Phe Leu Ala Gly Leu Asn Leu Lys Thr Ile Ile Ser 35 40 45 Leu Thr Pro Glu His Pro Ile Lys Pro Leu Leu Gln Phe Val Arg Thr 50 55 60 Ala Gly Ile Ser Phe Val His Leu Gly Leu Thr His Trp Arg Arg Pro 65 70 75 80 Gly Thr Asp Trp Arg Pro Val Arg Tyr Glu Ile Ile Lys Thr Ala Leu 85 90 95 Glu Ala Tyr Ile Leu Asp Thr Arg Ala His Pro Val Leu Leu Ile Asp 100 105 110 Pro Leu Gly Val His Gln Thr Gly Cys Leu Val Gly Ala Leu Arg Met 115 120 125 Met Gln Gly Trp Asn Phe Ala Ser Ala Leu Met Glu Tyr Arg Ala His 130 135 140 Ala Gly Ser Lys His Arg Tyr Leu Asp Glu Gln Tyr Ile Glu Leu Phe 145 150 155 160 Asp Ser Asp Leu Ile Asn Leu Pro Ala Pro Gln Tyr Arg Pro Ser Trp 165 170 175 Trp Leu Ser Cys Glu Glu Ala Asp Pro Gln Glu Val Lys Ala Leu Ala 180 185 190 Ser Ser Ser Gly Gly Thr Gly Leu Leu Ala Asp Thr Asn Gly Arg Thr 195 200 205 Gln Ala Ile Val 210 <210> 7 <211> 320 <212> PRT <213> artificial sequence <220> <223> SIT4 >CNAG_01436 Transcript 1 <400> 7 Met Arg Tyr Thr Ser Pro Thr Met Pro Ile Pro Ile Ser Ser Asp Pro 1 5 10 15 Asp His Trp Ile Gln His Ile Arg Gln Cys Lys His Leu Pro Glu Arg 20 25 30 Gln Met Lys Leu Leu Cys Asn Arg Val Arg Asp Leu Leu Leu Glu Glu 35 40 45 Ser Asn Val Arg Leu Val Gln Ser Pro Val Thr Val Cys Gly Asp Ile 50 55 60 His Gly Gln Phe Trp Asp Val Leu Glu Ile Phe Arg Gln Gly Gly Glu 65 70 75 80 Val Pro Lys Thr Ser Tyr Ile Phe Met Gly Asp Phe Val Asp Arg Gly 85 90 95 Tyr Tyr Ser Leu Glu Thr Leu Ser Leu Leu Leu Ala Tyr Lys Ala Arg 100 105 110 Tyr Pro Asp Lys Ile Thr Leu Leu Arg Gly Asn His Glu Ser Arg Gln 115 120 125 Ile Thr Gln Val Tyr Gly Phe Tyr Asp Glu Cys Met Gln Lys Tyr Gly 130 135 140 Asn Pro Ser Val Trp Lys Ala Cys Cys Asn Val Phe Asp His Leu Asn 145 150 155 160 Leu Ala Ala Ile Ile Asp Ser Ser Ile Leu Cys Val His Gly Gly Leu 165 170 175 Ser Pro Asp Ile Arg Thr Leu Asp Gln Ile Arg Thr Ile Ser Arg Ala 180 185 190 Gln Glu Val Pro His Glu Gly Ala Phe Cys Asp Leu Met Trp Ser Asp 195 200 205 Pro Asp Glu Val Glu Thr Trp Ser Ile Ser Pro Arg Gly Ala Gly Trp 210 215 220 Leu Phe Gly Gly Lys Val Thr Ser Glu Phe Asn Tyr Ile Asn Gly Leu 225 230 235 240 Ser Leu Ile Ala Arg Ala His Gln Leu Val Gln Glu Gly Tyr Lys His 245 250 255 Met Phe Asp Glu Ser Leu Val Thr Val Trp Ser Ala Pro Asn Tyr Cys 260 265 270 Tyr Arg Cys Gly Asn Ala Ala Ser Ile Met Gln Val Asp Glu Asp Gly 275 280 285 Arg Thr Ser Phe Lys Val Tyr Asp Ala Ala Ile Glu Asn Ser Thr Asp 290 295 300 Gln Lys Asn Pro Ala Met Arg Arg Val Gly Ala Pro Ser Tyr Phe Val 305 310 315 320 <210> 8 <211> 544 <212> PRT <213> artificial sequence <220> <223> GUA1 >CNAG_01877 Transcript 1 <400> 8 Met Ala Thr Glu Glu Ile His Ser Leu Tyr Asp Thr Ile Leu Ile Leu 1 5 10 15 Asp Phe Gly Ser Gln Tyr Ser His Leu Ile Thr Arg Arg Cys Arg Glu 20 25 30 Leu Asn Val Tyr Cys Glu Met Leu Pro Cys Thr Gln Lys Ile Ser Glu 35 40 45 Leu Ser Trp Lys Pro Lys Gly Ile Ile Leu Ser Gly Ser Pro Tyr Ser 50 55 60 Val Tyr Ala Pro Asp Ala Pro His Val Asp Pro Asp Val Phe Thr Leu 65 70 75 80 Gly Val Pro Ile Leu Gly Ile Cys Tyr Gly Leu Gln Glu Ile Ala Arg 85 90 95 Val His Gly Gly Thr Val Asp Ala His Thr His Arg Glu Tyr Gly Tyr 100 105 110 Ala Lys Ile Glu Val Val Lys Thr Gly Lys Lys Asp Gln Asp Ala Leu 115 120 125 Phe Glu Gly Ile Glu Met Glu Ala Asp Gly Gly Leu Gln Val Trp Met 130 135 140 Ser His Gly Asp Gln Leu Thr Ser Leu Pro Pro Asn Phe Val Thr Ile 145 150 155 160 Ala Ser Thr Pro Thr Ser Pro Phe Thr Ser Val Ala His Glu Ser Lys 165 170 175 Pro Ile Tyr Gly Val Gln Phe His Pro Glu Val Ser His Ser Pro Arg 180 185 190 Gly Lys Glu Val Ile Ala Ala Phe Val Lys Asn Val Cys Gly Val Arg 195 200 205 Asp Gly Trp Ser Met Glu Ser Phe Ile Pro Lys Glu Ile Ala Arg Ile 210 215 220 Arg Gln Ile Cys Gly Glu Lys Gly Gln Val Ile Gly Ala Val Ser Gly 225 230 235 240 Gly Val Asp Ser Thr Val Ala Ala Lys Leu Met His Glu Ala Ile Gly 245 250 255 Asp Arg Phe His Ala Ile Met Val Asp Asn Gly Val Leu Arg Lys Asp 260 265 270 Glu Ala Lys Lys Val His Lys Met Leu Thr Val Asp Leu Gly Val Asn 275 280 285 Leu Thr Val Ile Asp Ala Ser Glu Leu Phe Leu Ala Arg Leu Lys Gly 290 295 300 Val Glu Asp Pro Glu Arg Lys Arg Lys Ile Ile Gly Asn Thr Phe Ile 305 310 315 320 Glu Val Phe Glu Ala Glu Ala Ala Lys Leu Glu Ala Ala Ala Glu Lys 325 330 335 Glu Leu Ala Glu Lys Gly Gly Glu Ala Lys Gly Lys Ile Glu Trp Leu 340 345 350 Leu Gln Gly Thr Leu Tyr Pro Asp Val Ile Glu Ser Ile Ser Phe Lys 355 360 365 Gly Pro Ser Ala Thr Ile Lys Thr His His Asn Val Gly Gly Leu Leu 370 375 380 Glu Asp Met Lys Leu Lys Leu Ile Glu Pro Leu Arg Glu Leu Phe Lys 385 390 395 400 Asp Glu Val Arg Ala Leu Gly Arg Leu Leu Asn Ile Pro Glu His Leu 405 410 415 Val Gly Arg His Pro Phe Pro Gly Pro Gly Leu Ala Ile Arg Ile Leu 420 425 430 Gly Glu Val Thr Arg Glu Gln Ile Ala Ile Leu Gln His Ala Asp Asp 435 440 445 Ile Tyr Ile Glu Glu Ile Arg Ala Ala Gly Leu Tyr Asp Gln Ile Ser 450 455 460 Gln Ala Phe Val Ala Leu Leu Pro Val Lys Ala Val Gly Val Ala Gly 465 470 475 480 Asp Ala Arg Thr Tyr Asp Gln Val Val Ala Val Arg Ala Val Ser Thr 485 490 495 Glu Asp Phe Met Thr Ala Asp Trp Phe Val Phe Pro Pro Gln Val Leu 500 505 510 Lys Arg Ile Ser Ser Arg Ile Thr Asn Glu Val Lys Gly Val Asn Arg 515 520 525 Val Val Tyr Asp Ile Thr Ser Lys Pro Pro Gly Thr Val Glu Trp Leu 530 535 540 <210> 9 <211> 1344 <212> PRT <213> artificial sequence <220> <223> INP5201 >CNAG_02109 Transcript 1 <400> 9 Met Ala Ser Pro Leu His Glu Asn Asp Glu Glu Arg Pro Gln Ser Ile 1 5 10 15 Ala Ala Leu Arg Ser Lys Phe Glu Ser Leu Ala Ile Ala Gly Val Ser 20 25 30 Pro Ala Pro Thr Asp Val Pro Ser Ala Thr Asn Gly His Ala Thr Val 35 40 45 Ser Ser Ile Arg Asn Gly Leu Leu Ser Pro Arg Pro Glu Thr Pro Val 50 55 60 Asp Gly Gln Lys Ala Lys Pro Val Pro Pro Pro Lys Pro Ala Ser Arg 65 70 75 80 Pro Val Ser Pro Ala Thr Thr Ser Pro Ala Pro Gln Pro Ser Ser Leu 85 90 95 Leu Pro Pro Pro Ala Pro Arg Gln Ala Pro Ser Arg Pro Thr Thr Pro 100 105 110 Lys Pro Ser Phe Gln Thr His His Ser Thr Ser Ser Val Thr Ser Ile 115 120 125 Val Ser Ala Ala Ser Asp Ser His Leu Lys Pro Ser Asp Thr Met Ala 130 135 140 Ser Pro Pro Ala Val Ile Ser Pro Ala Val Ser Pro Ala Pro Thr Pro 145 150 155 160 Leu Arg Lys Ser Ala Pro Ser Val Pro Ser Lys Pro Pro Ser Val Ala 165 170 175 Val Thr Pro Ser Gly Ser Asp Gly Asp Glu Asp Glu Pro Val Ile Thr 180 185 190 Ser Val Lys Ala Leu Arg Glu Lys Phe Ser Gly Gln Ala Gln Ala Ser 195 200 205 Glu Ile Ala Leu Arg Lys Pro Val Asp Val Pro Lys Ala Ser Ala Val 210 215 220 Ser Val Val Lys Ala Ala Thr Val His Asp Ser Pro Glu Pro Leu Cys 225 230 235 240 Ala Pro Ser Ala Thr Pro Ile Pro Ala Pro Ile Pro Ala Pro Val Ile 245 250 255 Gln Arg Thr Leu Asp Gly Lys Thr Ser Pro Val Met Leu Ser Pro Ala 260 265 270 Ser Glu Gly Glu Ala Leu Ser Asp Thr Asn Asp Tyr Ser Ser His Pro 275 280 285 Thr Ala Pro Leu Ala Pro Pro Ala Pro Pro Ala Pro Ile Ser Arg Ile 290 295 300 Ser Ser Pro Val Pro Ala Pro Ala Pro Ala Pro Ser Gly Pro Pro Pro 305 310 315 320 Ile Asn Arg Ala His Lys Pro Pro Pro Arg Thr Ala Ile Ser Pro Ala 325 330 335 Pro Ile Phe Arg Pro Glu Ser Asn Val Ile Thr Pro Asn Thr Thr Ser 340 345 350 Pro Pro Ile Pro Gly Asn Lys Pro Val Ile Pro Ser Arg Ser Ser Ser 355 360 365 Ala Pro Glu Ala Ala Val Pro Pro Pro Pro Glu Arg Pro Gln Pro 370 375 380 Pro Gln Leu Pro Val Arg Arg Pro Thr Phe Ser Ser Pro Asp Thr Leu 385 390 395 400 Glu Pro Ser Thr Ala Ser Val Ile Ser Pro Pro Ala Leu Ala Ser Thr 405 410 415 Pro Leu Leu His Thr Ile His Asp Asp Thr Ala Leu Ala Pro Thr Pro 420 425 430 Ala Pro Ala Thr Ala Pro Pro Pro Leu Pro Asp Arg Ser Arg Ala Asn 435 440 445 Thr Ile Asn Arg Ser Glu Ser Glu Ser Ser Ala Thr Thr Thr Gly Pro 450 455 460 Pro Pro Pro Arg Leu Pro Ala Arg His Ala Ala Ile Pro Val Ser Ala 465 470 475 480 Gly Ser Gly Ser Thr Ser Ser Asn Ala Asn Gly Ser Gly Ser Thr Thr 485 490 495 Met Asn Pro Pro Pro Pro Pro Ala His Pro Ala Ser Pro Ser Lys Thr 500 505 510 Arg Ile Asn Ser Gly Gly Pro Pro Pro Pro Leu Leu Arg Ser Ala Thr 515 520 525 Val Asn Arg Gly Ser Ser Val Gly Ser Gly Ser Gly Ser Gly Gly Gly 530 535 540 Gly Gly Gly Ser Pro Pro Arg Arg Ser Asn Thr Ile Ser Arg Ala Ala 545 550 555 560 Pro Phe Thr Gln Glu Lys Tyr Ser Thr Ser Ala Thr Ser Leu Gly Leu 565 570 575 Gly Glu Lys Gly Val Tyr Ser Asp Glu Asp Asp Glu Pro Glu Glu Pro 580 585 590 Gly Ala Val Thr Asn Leu Ser Ala Gln Ala Lys Arg Met Leu Asp Glu 595 600 605 Phe Pro Asp Met Thr Glu Ala Asn Arg Arg Pro Pro Val Phe Val Pro 610 615 620 Asp Ile Arg Val Lys Glu Cys His His Val Ser Ala Phe Ala Val Tyr 625 630 635 640 Gly Arg Tyr Val Cys Thr Gly Ala His His Val Arg Val Tyr Asp Thr 645 650 655 Gln Leu Ser Asp His Ala Ile Ser Val Val Asp Leu Lys Glu Thr Gly 660 665 670 Leu Glu Ser Arg Gly Lys Asp Pro Lys Val Thr Ala Met Cys Phe Arg 675 680 685 Pro Gly Ala Thr Glu Ser Glu Glu Gly Arg Tyr Leu Trp Cys Gly Thr 690 695 700 Lys Asp Gly His Leu Trp Glu Leu Asp Ile Ser Thr Gly Glu Val Thr 705 710 715 720 Ser Thr Lys Ala Phe Val His Thr Ser Ser Ile Ser Tyr Ile Trp Arg 725 730 735 His Arg Lys Asn Ile Ile Ser Leu Asp Glu Gly Gly Lys Leu Leu Val 740 745 750 Phe Asp Val Gly Asp Ile Glu Gly Lys Pro Pro Thr Met Ala Arg Gln 755 760 765 Leu Arg Ile Gly Asp Lys Phe Gly Phe Ala Lys Leu Ile Cys Gly Lys 770 775 780 Leu Trp Thr Ser Ser Gly Pro Leu Thr Arg Ser Thr Thr Ser Ser Ala 785 790 795 800 Thr Ser Lys Gly Pro Thr Val Arg Ile Tyr Asp Pro Cys Ala Pro Gly 805 810 815 Thr Met Pro Pro Pro Lys Thr Ile Phe Ala Thr Glu Trp Ala Gly Ala 820 825 830 Val Thr Ser Ala Thr Tyr Met Pro Leu His His Asp Thr Ile Phe Leu 835 840 845 Gly His Glu Gly Gly Phe Val Ser Val Trp Asp Gly Lys Glu Leu Val 850 855 860 Cys Lys Gln Val Leu Lys Ile Ser Ser Thr Asp Val Leu Ala Leu Glu 865 870 875 880 Gly Val Gly Glu Tyr Leu Trp Thr Gly Asn Arg Lys Gly Gln Ile His 885 890 895 Val Phe Asp Ile Lys Glu Lys Pro Trp Leu Ala Thr Asn Ile Trp Ile 900 905 910 Gly His Pro Asp Asn Pro Val Gln Ser Leu Val Val Asp Pro Tyr Ser 915 920 925 Ile Gln Ser Ala Gly Arg Tyr Thr Cys Trp Ser Phe Ala Arg Asp Ala 930 935 940 Leu Arg Ala Trp Asp Gly Leu Leu Ser Val Asp Trp Ile Asp Lys Gln 945 950 955 960 Leu Thr Ala Arg Gln Ser Ser Phe Cys Thr Phe Arg Pro Val Asn Val 965 970 975 Leu Ile Cys Thr Trp Asn Ile Asp Ser Ala Lys Pro Thr Asp Leu Asn 980 985 990 Gly Ser Val Ala Asn Ala His Phe Leu Glu Asp Val Leu Arg Ser Val 995 1000 1005 Asp Ser Pro Asp Ile Ile Val Phe Gly Phe Gln Glu Val Ile Pro Leu 1010 1015 1020 Thr Asp Lys Lys Tyr Thr Ala Lys Thr Leu Leu Phe Gly Asn Lys Ser 1025 1030 1035 1040 Lys Asp Gly Gly Ala Ala Ala Asp Arg Val Ser His Ala Tyr Arg His 1045 1050 1055 Trp Leu Glu Lys Leu Gln Ser Ala Val Gln Met Ala Ser Pro Ser Asn 1060 1065 1070 Cys Pro Tyr Ile Lys Ile His Ser Glu Ser Leu Val Gly Leu Phe Thr 1075 1080 1085 Cys Ile Phe Val Lys Gln Ser Glu Lys Ile Ser Leu Arg Asp Leu Asp 1090 1095 1100 Ile Thr Thr Val Lys Arg Gly Ile Gly Gly Ile Tyr Gly Asn Lys Gly 1105 1110 1115 1120 Ala Ile Val Ser Arg Leu Val Met Asp Asp Thr Ser Ile Cys Phe Ile 1125 1130 1135 Asn Val His Leu Ala Ala Gly Gln Ser Gln Lys Ala Ser Arg Asn Ala 1140 1145 1150 Asp Leu Ala Gly Ile Leu Glu Asp Lys Ala Ile Phe Pro Pro Ala Asp 1155 1160 1165 Glu Leu Pro Phe Val His Gly Gly Cys Gly Thr Gly Ile Leu Asp His 1170 1175 1180 Glu Met Val Phe Leu Asn Gly Asp Leu Asn Tyr Arg Ile Asp Gln Arg 1185 1190 1195 1200 Arg Glu Asn Val Ile Ser Ser Ile Ala Asn Gly Glu Leu Ala Tyr Leu 1205 1210 1215 Leu Glu His Asp Gln Leu Arg Lys Glu Met Arg Thr Asn His Ala Phe 1220 1225 1230 Arg Leu Arg Asn Phe Glu Glu Ala Pro Ile Thr Phe Ala Pro Thr Tyr 1235 1240 1245 Lys Tyr Asp Pro Gly Thr His Asp Tyr Asp Ser Ser Glu Lys Arg Arg 1250 1255 1260 Ile Pro Ala Trp Cys Asp Arg Ile Leu Tyr Lys Lys Ser Pro Arg Val 1265 1270 1275 1280 Gln Ala Leu Asn Tyr Gln Arg Tyr Glu Pro Thr Val Ser Asp His Arg 1285 1290 1295 Pro Val Ser Ala Gly Tyr Thr Ile Ile Leu Lys Ala Ile Asp Ser Leu 1300 1305 1310 Lys Met Met Asp Val Arg Arg Glu Ala Thr Gly Glu Trp Ala Lys Arg 1315 1320 1325 Glu Lys Glu Leu Leu Glu Lys Met Gln Glu Val Phe Asp Gly Ile Glu 1330 1335 1340 <210> 10 <211> 332 <212> PRT <213> artificial sequence <220> <223> PHS1 >CNAG_02487 Transcript 1 <400> 10 Met Phe Pro Ser Asp Thr Lys Ala Ser Leu His Ser Ala Phe Gln Ser 1 5 10 15 Val Leu Asp Ser Ala Ser Pro Ser Ile Leu Ala Arg Leu Ser Leu Gln 20 25 30 Leu Ala Ala Met Pro His Val Ala Ala Pro Ser Arg Gln Ala Leu Glu 35 40 45 Gln Glu Arg Leu His Arg His Ala Gln Gln Pro Gly Ser Ser Leu Ser 50 55 60 Pro Ala Pro Arg Leu Ser Pro Leu Arg Leu Tyr Leu Leu Gly Tyr Asn 65 70 75 80 Ile Leu Ser Ala Leu Leu Trp Gly His Leu Leu Val Leu Thr Leu Ser 85 90 95 Phe Leu Leu Ala Pro Ser Arg Pro Pro Trp His Gln Leu Ala Asp Arg 100 105 110 Leu Ser Gly Ser Tyr Asp Tyr His Asn Leu Gly Trp Cys Thr Lys Trp 115 120 125 Thr Gln Thr Leu Ala Val Leu Glu Val Val His Ala Ala Leu Gly Trp 130 135 140 Val Arg Ser Pro Leu Gly Thr Val Ala Ser Gln Val Ala Ser Arg Leu 145 150 155 160 Trp Thr Val Trp Gly Val Val Glu Ala Ala Pro Glu Ile Thr His Gly 165 170 175 His Pro Leu Phe Thr Thr Met Leu Leu Ala Trp Ser Leu Thr Glu Val 180 185 190 Ile Arg Tyr Ser Phe Tyr Ala Leu Ser Leu Leu Ser Val Ser Ala Pro 195 200 205 Phe Leu Asn Tyr Leu Arg Tyr Thr Thr Phe Ile Pro Leu Tyr Pro Leu 210 215 220 Gly Ala Ser Ser Glu Ala Phe Leu Ser Phe Ala Thr Leu Pro Ala Leu 225 230 235 240 Ala Pro Val Val Ser Arg Ala Val Thr Asn Val Met Ala Gln Ala Pro 245 250 255 Arg Glu Ile Met Lys Thr Lys Val Gly Arg Glu Val Leu Trp Trp Ser 260 265 270 Ala Lys His Gly Gly Gly Thr Ala Gly Ala Gln Lys Glu Trp Gly Trp 275 280 285 Ile Glu Ile Val Arg Ala Gly Leu Phe Leu Leu Trp Trp Pro Ala Leu 290 295 300 Tyr Val Leu Tyr Thr Tyr Met Leu Lys Gln Arg Arg Lys Val Leu Gly 305 310 315 320 Lys Gly Lys Thr Val Gly Gly Val Ser Lys Ala Gln 325 330 <210> 11 <211> 721 <212> PRT <213> artificial sequence <220> <223> MRE11 >CNAG_02490 Transcript 1 <400> 11 Met Ser Ala Pro Asn Arg Val Pro Asp Ser Gln Pro Ser Ser Glu Ile 1 5 10 15 Gly Asp Glu Pro Pro Ser Ile Val Glu Pro Asp Leu Glu Asn Cys 20 25 30 Phe Arg Ile Leu Ile Ala Thr Asp Asn His Ile Gly Tyr Ala Glu Lys 35 40 45 Asp Pro Val Arg Gly Gln Asp Ser Ile Asn Thr Phe Arg Glu Ile Leu 50 55 60 Glu Leu Ala Arg Asp His Asp Val Asp Phe Ile Leu Leu Ala Gly Asp 65 70 75 80 Leu Phe His Glu Asn Arg Pro Ser Arg Thr Cys Met His Gln Thr Ile 85 90 95 Ala Leu Leu Arg Glu Phe Thr Leu Gly Asp Lys Pro Ile Glu Phe Glu 100 105 110 Leu Leu Ser Asp Pro Met Asp Gly Ser Thr Pro Gly Phe Ser Phe Pro 115 120 125 Ala Val Asn Tyr Glu Asp Pro Asn Ile Asn Ile Ala Ile Pro Val Phe 130 135 140 Ser Ile His Gly Asn His Asp Asp Pro Gln Gly Thr Gly Pro Glu Gly 145 150 155 160 Ala Leu Cys Ala Leu Asp Val Leu Ser Val Ser Gly Val Leu Asn Tyr 165 170 175 Phe Gly Lys Ser Asp Leu Val Ala Asp Glu Ser Ala Ala Asp Asn Pro 180 185 190 Glu Lys Gly Ile His Ile Arg Pro Val Leu Leu Arg Lys Gly Thr Thr 195 200 205 His Val Ala Leu Tyr Gly Cys Gly Asn Ile Arg Asp Gln Arg Met Tyr 210 215 220 Gln Glu Leu Arg Ala Asn Lys Val Lys Met Phe Met Pro Thr Gly Gly 225 230 235 240 Asp Val Pro Asp Ser Glu Trp Phe Asn Ile Leu Leu Val His Gln Asn 245 250 255 Arg Val Arg His Gly Pro Gln Asn Tyr Val Pro Glu Asn Met Phe Asp 260 265 270 Asp Ser Met Arg Leu Val Ile Trp Gly His Glu His Asp Cys Arg Ile 275 280 285 Thr Pro Glu Ser Val Ala Asp Lys Asn Tyr Phe Ile Thr Gln Pro Gly 290 295 300 Ser Ser Val Ala Thr Ser Leu Ala Pro Gly Glu Ala Val Pro Lys His 305 310 315 320 Val Gly Leu Leu Ser Ile Gln Gly Ser Gln Phe Gln Leu Glu Glu Leu 325 330 335 Pro Leu Lys Thr Val Arg Pro Phe Glu Leu Asp Glu Val Val Leu Ser 340 345 350 Tyr Ala Ala Glu Gln Gly Ala Val Asp Leu Asn Asp Arg Asp Ser Ile 355 360 365 Thr Ser Phe Leu Arg Glu Gln Val Glu Ala Leu Ile Leu Gln Ala Lys 370 375 380 Lys Asn Trp Lys Glu Arg Asn Asn Gly Ser Thr Lys Asn Met Met Leu 385 390 395 400 Pro Leu Ile Arg Leu Lys Val Glu Thr Thr Asp Ala Lys Glu Met Val 405 410 415 Asn Pro Val Arg Phe Gly Gln Glu Tyr Val Asn Arg Val Ala Asn Pro 420 425 430 Arg Asp Ile Leu Gln Tyr Tyr Arg Lys Lys Lys Asn Glu Arg Lys Val 435 440 445 Lys Asn Asn Pro Asp Met Pro Asn Ile Asn Asp Asp Glu Trp Glu Glu 450 455 460 Asp Pro Glu Ser Leu Thr Ala Asp Glu Arg Leu Ser Lys Leu Arg Met 465 470 475 480 Ala Thr Leu Val Lys Gln Tyr Leu Gln Ala Gln Ser Leu Asp Val Leu 485 490 495 Val Glu Asn Gly Met Glu Asp Ala Val Met Arg Phe Val Asp Lys Asp 500 505 510 Asp Lys Asp Ala Ile Lys Asp Phe Val Ala Asp Thr Leu Arg Met Val 515 520 525 Gly Arg Lys Met Lys Glu Arg Glu Val Lys Glu Asp Asp Val Asp Leu 530 535 540 Ala Met Ala Glu Ala Lys Glu Lys Glu Tyr Asn Arg Tyr Ala Asp Ser 545 550 555 560 Asn Pro Val Pro Ser Gln Ser Val Lys Gly Lys Asn Lys Gln Arg Asp 565 570 575 Ser Asp Val Asp Ser Met Met Ala Ser Asp Asp Asp Met Asp Met Asp 580 585 590 Glu Met Pro Thr Gln Gln Arg Ala Pro Val Arg Arg Ala Thr Ala Asn 595 600 605 Gln Pro Val Arg Ser Ala Lys Gly Lys Gly Lys Gln Pro Leu Phe Glu 610 615 620 Asn Ala Ser Glu Glu Glu Glu Asp Glu Glu Glu Glu Glu Glu Glu Glu 625 630 635 640 Glu Glu Pro Ala Pro Lys Lys Gly Arg Gly Arg Ala Ala Ala Ala Ser 645 650 655 Thr Lys Lys Ala Pro Ala Lys Lys Pro Pro Ala Arg Thr Pro Ala Lys 660 665 670 Ser Thr Thr Lys Ala Pro Ala Gly Arg Arg Pro Ala Val Ser Gln Pro 675 680 685 Ser Thr Gly Arg Gly Val Thr Gln Ser Gln Leu Thr Phe Ser Arg Ser 690 695 700 Gly Thr Gly Lys Ala Ala Ala Val Pro Ile Glu Leu Ser Ser Asp Glu 705 710 715 720 Asp <210> 12 <211> 605 <212> PRT <213> artificial sequence <220> <223> DBR1 >CNAG_03222 Transcript 1 <400> 12 Met Arg Ile Ala Ile Gln Gly Cys Ser His Gly Ser Leu Ala Gln Ile 1 5 10 15 Tyr Asp Val Val Asn Tyr Tyr Ser Ser Gln Thr Lys Asn Pro Ile Asp 20 25 30 Leu Leu Leu Leu Cys Gly Asp Phe Gln Ala Leu Arg Ser Lys His Asp 35 40 45 Tyr Ala Ser Leu Ala Val Pro Ala Lys Phe Lys Gln Leu Gly Ser Phe 50 55 60 His Gln Tyr Tyr Ser Gly Glu Arg Val Ala Pro Val Leu Thr Ile Val 65 70 75 80 Ile Gly Gly Asn His Glu Ala Ser Asn Tyr Met Trp Glu Leu Tyr His 85 90 95 Gly Gly Trp Leu Ala Pro Ser Ile Tyr Tyr Leu Gly Ala Ala Gly Ser 100 105 110 Val Tyr Val Asn Gly Val Arg Ile Val Gly Ala Ser Gly Ile Tyr Lys 115 120 125 Gly Phe Asp Tyr Arg Lys Gly His Phe Glu Lys Val Pro Tyr Asn Asp 130 135 140 Lys Glu Leu Arg Ser Val Tyr His Ile Arg Glu Tyr Asp Val Glu Lys 145 150 155 160 Leu Met His Leu Thr Pro Ser Pro Ser Thr Ile Phe Leu Ser His Asp 165 170 175 Trp Pro Thr Thr Ile Ala His His Gly Asn Lys Asn Ala Leu Leu Lys 180 185 190 Arg Lys Pro Phe Phe Arg Asp Glu Ile Glu Lys Asn Thr Leu Gly Ser 195 200 205 Pro Pro Leu Leu Arg Leu Met Asn His Phe Gln Pro Ser Tyr Trp Phe 210 215 220 Ser Ala His Leu His Val Lys Phe Ala Ala Leu Tyr Glu His Gln Ala 225 230 235 240 Pro Asn His Gly Pro Asp Val Asp Gly Gly Ala Pro Leu Pro Leu Leu 245 250 255 Ala Thr Ser Ala Val Ile Ala Gln Ala Gly Gly Asn Pro Asp Glu Ile 260 265 270 Gln Ile Asp Glu Glu Met Asp Ala Gly Asn Pro Asp Glu Ile Ile Val 275 280 285 Glu Asp Glu Gly Glu Glu Val Ile Ile Arg Pro Arg Gln Val Asn Pro 290 295 300 Asp Glu Ile Ala Met Asp Asp Asp Glu Phe Asp Asp Pro Ala Pro Ala 305 310 315 320 Val Pro Gln Pro Leu Pro Ala Thr Thr Asn Ser Ala Phe Asn Pro Glu 325 330 335 Glu Ile Ile Ile Ser Asp Gln Glu Phe Asp Ala Pro Thr Thr Val Ser 340 345 350 Gln Pro Leu Gln Pro Leu Pro Pro Thr Lys Thr Asn Ala Ser Asn Pro 355 360 365 Glu Glu Ile Ala Ile Ser Asp Asp Glu Phe Asp Asp Pro Ala Pro Leu 370 375 380 Ala Gln Ser Leu Thr Thr Ile Asp Glu Ser Thr Asp Leu Ile Ala Gln 385 390 395 400 Ser Arg Ser Asn Pro Ser His Pro Pro Val Ala Gly Thr Ile Ala Pro 405 410 415 Pro Thr Ser Asp Ser Ile Ala Ser Arg Val Met Gln Glu Ala Arg Gln 420 425 430 Glu Gln Gln Lys Trp Glu Leu His Gly Gly Lys Gly Met Glu Gly Val 435 440 445 Thr Lys Phe Leu Ala Leu Asp Lys Cys Gly Pro Gly Lys Asp His Met 450 455 460 Gln Phe Leu Glu Ile Pro Asp Pro Ser Pro Pro Pro Ile Pro Gly Pro 465 470 475 480 Pro Arg Leu Thr Tyr Asp Pro Glu Trp Leu Ala Ile Ser Arg Ala Phe 485 490 495 His Pro Tyr Leu Ser Thr Ser Tyr Gln Pro Ile Pro Leu Pro Ser Ser 500 505 510 Asp Ile Leu Glu Gln Met Val Lys Asp Glu Val Ala Arg Ile Lys Glu 515 520 525 Glu Gly Leu Leu Val Pro Thr Val Pro Gln Asp Gly Ala Val Glu Gly 530 535 540 Gln Glu Gly Leu Val Trp Glu Lys Gly Lys Val Asp Val Gly Arg Val 545 550 555 560 Gln Arg Phe Trp Trp Thr Ala Pro Pro Glu Gly His Pro Gly Gly Asn 565 570 575 Asp Thr Ala Trp Tyr Thr Asn Pro Gln Thr Glu Ala Phe Cys Gly Met 580 585 590 Leu Gly Val Gln Asn Lys Ile Asn Pro Pro Val Asn Arg 595 600 605 <210> 13 <211> 281 <212> PRT <213> artificial sequence <220> <223> SIW14 >CNAG_03296 Transcript 1 <400> 13 Met Met Ser Ser Ser Pro Thr Met Ser Ser Thr Pro Pro Gln Val Pro 1 5 10 15 Ser Phe Leu Ala Asn Ile Leu Leu Ser His Leu Ser Pro Leu Asp Pro 20 25 30 Ser Ser Thr Ser Pro Thr Thr Phe Glu His His Arg Ser Pro Thr Thr 35 40 45 Gln Asn Pro Ser His Pro Gln Ala Leu Gln Thr Ala Glu Pro Pro Pro 50 55 60 Asn Pro Leu Tyr Leu Pro Pro Pro Ala Leu Pro Lys Val Glu Glu Asp 65 70 75 80 Leu Val Pro Pro Glu Asn Phe Ala Leu Val Ser Ser Gly Val Tyr Arg 85 90 95 Cys Gly Phe Pro Lys Lys Arg Asn Phe Lys Phe Met Glu Thr Leu Arg 100 105 110 Leu Lys Thr Val Leu Thr Leu Val Leu Glu Glu Tyr Pro Lys Ala Asn 115 120 125 Leu Glu Trp Cys Gln Ser Gln Asp Ile Gln Phe Met Gln Phe Gly Ile 130 135 140 Pro Gly Asn Lys Glu Pro Phe Asp Asn Ile Pro Glu Asp Val Ile Cys 145 150 155 160 Ala Ala Leu Val Ala Ile Leu Asp Arg Arg Asn His Pro Ile Leu Ile 165 170 175 His Cys Asn Lys Gly Lys His Arg Thr Gly Cys Leu Ile Gly Cys Ile 180 185 190 Arg Arg Leu Gln Ala Trp Ser Leu Thr Ser Ile Phe Asp Glu Tyr Arg 195 200 205 Arg Phe Ser Ala Pro Lys Ser Arg Ala Val Asp Gln Gln Phe Ile Asp 210 215 220 Leu Phe Asp Ile Met Pro Val Trp Glu Ala Val Cys Arg Pro Lys Gly 225 230 235 240 Gly Gly Leu Gly Asn Leu Pro Asp Trp Gly Met Leu Val Leu Pro Lys 245 250 255 Gly Val Val Glu Val Gly Arg Asp Gly Lys Glu Lys Lys Arg Val Glu 260 265 270 Arg Asp Ile Leu His Met Arg Gly Leu 275 280 <210> 14 <211> 881 <212> PRT <213> artificial sequence <220> <223> SDP102 >CNAG_03893 Transcript 1 <400> 14 Met Gln Pro Gln Gln His Thr His Asn Asn Pro Ser Pro Ala Ala Arg 1 5 10 15 Pro Gln Pro Leu Arg Val Val His Ser Pro Thr Ile Pro Pro Pro Asn 20 25 30 Arg Gly Thr Arg Thr Ser Glu Leu Ala Gln Gly Pro Lys Ala Pro Leu 35 40 45 Ala Ala Pro Leu Pro Leu Arg Ile Thr Ala Ala Thr Ser Pro Asn Arg 50 55 60 Lys Arg Pro Thr Pro Leu Val Leu Gly Lys Pro Arg Glu Ala Gly Pro 65 70 75 80 Glu Ala Gly Pro Glu Asp Trp Glu Ile His Gln Glu Ile Ser Phe Ala 85 90 95 Ala Ser Leu Gly Ala Ser Ala Asp His Ser Leu Asp Asn Glu Leu Gln 100 105 110 Asp Leu Ser Lys Leu Arg Lys Ala Val Arg Gln Asn Leu Leu Ala Arg 115 120 125 Pro Ile Asp Ser Pro Leu Glu Leu Ser Gly Ser Asp Gln Ser Ala Phe 130 135 140 Asn Thr Pro Gly Gln Gln Ser Ser Phe Gly Ser Ser Ser Ile Ser Met 145 150 155 160 Asp Ser Ile Pro Ile Glu Gln Val Phe Asp Arg Val Glu Gly Gly Ser 165 170 175 Val Leu Leu Val Asp Thr Arg Pro Leu Ala Ser Phe Leu Asn Ser His 180 185 190 Leu Pro Asn Ser Ile Pro Leu Ser Val Pro Thr Leu Leu Ser Lys Arg 195 200 205 Phe Gln Lys Ser Gln Ser Gln Ser Ser Pro Ser Ser Ile Ser Trp Ala 210 215 220 Thr Leu Ser Pro Phe Val Ser Leu Ser Ser Ala Arg Glu Arg Trp Asp 225 230 235 240 Ser Val Asp Gln Asp Lys Val Glu Ile Ala Val Ile Cys Gln Gly Glu 245 250 255 Glu Gly Arg Val Val Lys Glu Ile Leu Lys Ser Leu Ile Glu Gly Arg 260 265 270 Val Lys Val Val Lys Gly Gly Trp Ala Ala Val Leu Asn Tyr Glu Arg 275 280 285 Ala Arg Arg Thr Leu Val Ser Gly Gln Thr Ala Thr Arg Pro Cys Leu 290 295 300 Asp Val Thr Ser Pro Glu Thr Asp Ser Lys Pro Leu Pro Pro Ala Ser 305 310 315 320 Ala Ser Asn Ile Leu Pro Pro Lys Ser Ala Pro Pro Cys Asp Ile Pro 325 330 335 Leu Pro Pro Ile Pro Ala Ser Pro Ser Pro Pro Lys Ser Leu Asn His 340 345 350 Arg Pro Ser Leu Pro Ser Leu Arg Pro Pro Phe Thr Gly Pro Thr Arg 355 360 365 Asn Leu Pro Ser Leu Ser Ile Asn Ala Gly Gln Ala Ser Gln Arg Arg 370 375 380 Thr Pro Lys Leu Ser Leu Asn Phe Asp Arg Pro Leu Lys Ser Ala Thr 385 390 395 400 Leu Gly Gly Tyr His Asp Ile Pro Pro Thr Pro His Gly Phe Ser Cys 405 410 415 Thr Arg Thr Arg Pro Gln Arg Ser Pro Gly Leu Ser Leu Asn Ile Pro 420 425 430 His Thr Pro Phe Gln Pro Gln Gln Gly Gln Ile Gln Asp Arg Ile Leu 435 440 445 Glu Asp Ser Arg Pro Asn Gly Ser Gly Ser Ile Gln Thr Lys Ala His 450 455 460 Glu Gln Ser Arg Phe Pro Pro Ser Ser Ser Thr Phe Gly Asp Ala Lys 465 470 475 480 Gln Ile Glu Asn Glu Gly Glu Asp Met Ala Pro Asn Leu Tyr Asp Gly 485 490 495 Pro Ala Pro Arg Ala Pro Thr Ser His Ser Pro Ser Lys Ser Gln Asp 500 505 510 Tyr Gln Ala Ala Arg Phe Tyr Ser Ser Pro Ser Ser Met Asn Ser Ala 515 520 525 Leu Pro Ala Ser Pro Pro Thr Thr Arg Pro Ala Val Ala Pro Phe Asn 530 535 540 Pro Ser Val Ile Leu Pro Ser Phe Leu Tyr Leu Gly Pro Asp Ile Gln 545 550 555 560 Ser Glu Ser Asp Val Gln Tyr Leu Phe Arg Leu Gly Val Lys Arg Ile 565 570 575 Leu Asn Val Ala Leu Glu Cys Asp Asp Asn Gln Gly Leu Ser Leu Lys 580 585 590 Glu Arg Phe Lys Tyr Arg Lys Val Gly Met Arg Asp Ile Val Glu Glu 595 600 605 Asn Gly Val Gly Lys Gly Met Arg Asp Ala Cys Glu Phe Leu Asp Asp 610 615 620 Ala Arg Leu His Ser Ala Pro Thr Tyr Val His Cys Gln Ala Gly Lys 625 630 635 640 Ser Arg Ser Val Thr Ile Ile Leu Ala Tyr Leu Ile His Ala Asn Ala 645 650 655 Trp Thr Leu Lys Thr Ser Tyr Ala Tyr Val Ala Glu Arg Arg Lys Gly 660 665 670 Ile Ser Pro Asn Ile Gly Phe Val Ala Glu Leu Met Gln Trp Glu Glu 675 680 685 Lys Glu Leu Gly Val Lys Gln Ser Gly Gly Val His Gly Asp Gly Asn 690 695 700 Gly Arg Ala Lys Ala Pro Gly Gly Gly Gly Gly Gly Gly Gly Ser Arg 705 710 715 720 His Met Glu Asp Gly Gly Asp Asp Glu Gly Lys Gly Lys Thr His Leu 725 730 735 Arg Asp Ser Leu Pro Pro Thr Trp Ser Ser Ser Val Asp Thr Tyr Thr 740 745 750 Arg Pro Ala Lys Val Tyr Ser Pro Val Gly Arg Asp Asp Gly Gly Glu 755 760 765 Glu Glu Ser Gly Arg Glu Gly Arg Ile Ala Val Gly Asp Glu Arg Glu 770 775 780 Val Arg Lys Asn Gly Val Trp Met His His Arg Arg Ala Pro Val Asp 785 790 795 800 Arg Thr Thr Leu Gln Pro Gly Arg Arg Val Ser Lys Ala Gly Leu Glu 805 810 815 Ser Leu Arg Pro Phe Leu Ile Thr Ser Thr Asp Ala Ser Ser Ser Ser 820 825 830 Ala Ala Pro Asn Asn Gly Asp Asn Ile Asp Ser Glu Arg Gln Val Asn 835 840 845 Asn Gly Ser Glu Ala Arg Pro Ser Pro Arg Ala Ser Pro Gly Met Gly 850 855 860 Met Gly Gly His Ala Met Thr Pro Ala Gly Asp Gly Pro Leu Lys Trp 865 870 875 880 Ile <210> 15 <211> 185 <212> PRT <213> artificial sequence <220> <223> OCA1 >CNAG_03963 Transcript 1 <400> 15 Met Ala Lys Ile Val Pro Pro Met Asn Phe Gly Leu Val Glu Asp Gly 1 5 10 15 Phe Tyr Arg Ser Ala Gln Pro Ser Glu Leu Cys Phe Ser Phe Leu Glu 20 25 30 Lys Leu Asn Leu Lys Ser Ile Ile Trp Val Gly Ala Glu Glu Pro Ser 35 40 45 Asp Ile Phe Leu Ser Phe Ile Glu Ser Gln Gly Ile Lys Leu Tyr Asn 50 55 60 Leu Ala Pro Gln Thr Ser Leu Asn Pro His Phe Pro Pro Pro Tyr Thr 65 70 75 80 Asp Ser Gly Val Val Pro Ile Ser Gly Gln Tyr His Leu Pro Pro Leu 85 90 95 Pro Pro Pro Pro Glu Pro Leu Ile Ile Gln Ala Leu Thr Leu Leu Leu 100 105 110 Arg Pro Ser Thr Phe Pro Thr Leu Leu Cys Cys Asn Met Gly Arg His 115 120 125 Arg Thr Gly Thr Val Val Gly Cys Tyr Arg Lys Leu Gln Arg Trp Ala 130 135 140 Leu Ser Ser Ile Leu Glu Glu Tyr Arg Arg Tyr Ala Gly Met Lys Val 145 150 155 160 Arg Val Leu Asn Glu Gln Phe Ile Glu Leu Phe Asp Thr Asp Leu Val 165 170 175 Ser Ile Thr Ala Glu Gln Val Thr Lys 180 185 <210> 16 <211> 671 <212> PRT <213> artificial sequence <220> <223> GDA1 >CNAG_03996 Transcript 1 <400> 16 Met Pro Ala His Leu Phe Ala Arg Ser Thr Asp Thr Thr Phe Pro Ser 1 5 10 15 Ser Ile Glu Thr Ser Thr Ser Gly Ser Leu Ser Phe Ser Leu Leu Ser 20 25 30 Ser Leu Arg His Arg Pro Ser Gly Arg Ile Pro Lys Thr Pro Ile Ser 35 40 45 Pro Lys Ser Pro Thr Ser Ala Ser Thr Ser Thr Thr Ala Ala Met Phe 50 55 60 Ser Thr Arg Lys Tyr Ser Pro Leu Pro Thr Ser Ala Asn Gly Pro Ala 65 70 75 80 Arg Lys Arg Thr Gly Ala Gly Leu Thr Ala Trp Lys Arg Trp Ala Leu 85 90 95 Leu Ala Ala Ile Ser Val Ala Val Ile Phe Leu Val Phe Ser Arg Ala 100 105 110 Ser Gly Gly Ser Glu Gln Gln Gln Ile Tyr Asn Glu Glu Asn Thr Tyr 115 120 125 Thr Pro Ser Leu Asp Glu Asp Val Val Gly Asp Gly Asp Pro Ile Asp 130 135 140 Tyr Ser Ser Pro Pro Phe Arg Pro Glu Asp Ser Asp Val Ala Gln Pro 145 150 155 160 Leu Asp His Glu Asp Gly Asp Asp Asp Gly Val Ile His Thr Leu Pro 165 170 175 Thr Gly Asp Ala Ser Asn Pro His Asp Pro Thr Ser Thr Glu Ala Gln 180 185 190 Asp Ala Ser Glu Ala Glu Gln Asp Phe Thr Asn Glu Ser Glu Ser Glu 195 200 205 Ser Pro Ser Glu Ala Glu Ser Ser Phe Pro Gly Ser Phe Glu Gln Asp 210 215 220 Pro Asp Pro Ala Ser Thr Thr Ala Cys Thr Glu Pro Val Ser Ser Asp 225 230 235 240 Lys Pro Val Val Gln Tyr Ala Leu Thr Ile Asp Ala Gly Ser Thr Gly 245 250 255 Ser Arg Ile His Val Tyr Lys Phe Asn Asn Cys Gly Pro Ser Pro Gln 260 265 270 Leu Glu Tyr Glu Thr Phe Lys Ala Val Lys Pro Gly Leu Ser Ala Tyr 275 280 285 Ala Arg Asp Pro Thr Ala Ala Ala Ala Ser Leu Asp Pro Leu Leu Glu 290 295 300 Glu Ala Tyr Arg Val Val Pro Glu Ser Leu Arg Lys Cys Thr Pro Val 305 310 315 320 Glu Val Lys Ala Thr Ala Gly Leu Arg Leu Leu Gly Gln Gln Glu Ser 325 330 335 Glu Ala Ile Leu Asp Glu Val Arg Asn Arg Leu Glu Thr Asn Trp Asp 340 345 350 Phe Thr Val Ser Gly Glu Arg Ala Val Glu Ile Met Asp Gly Lys Asp 355 360 365 Glu Gly Val Tyr Ala Trp Ile Thr Ala Asn Tyr Leu Leu Asn Lys Ile 370 375 380 Gly Glu Gly Ala Glu Ser Asp Asp Thr Leu Ala Val Met Asp Leu Gly 385 390 395 400 Gly Ala Ser Thr Gln Ile Val Phe Glu Pro Lys Phe Pro Ala Glu Ser 405 410 415 Asp Gln Ala Leu Val Glu Gly Glu His Lys Tyr Glu Leu Thr Phe Gly 420 425 430 Gly Lys Asp Phe Thr Leu Tyr Gln His Ser Tyr Leu Gly Tyr Gly Leu 435 440 445 Met Arg Ala Arg Arg Ser Val His Asn Leu Val Ala Phe Thr Trp Ser 450 455 460 Phe Gly Gln Gly Glu Val Glu Trp Glu Asn Leu Ser Glu Asp Val Gln 465 470 475 480 Val Pro Asn Pro Cys Leu Ser Lys Gly Met Thr Arg Arg Val Ala Leu 485 490 495 Asp Pro Pro Gly Arg Gln Thr Val Asn Val Thr Met His Gly Gly Asn 500 505 510 Gly Asn Phe Glu Ala Cys Asn Arg Val Val Glu Leu Val Met Ala Lys 515 520 525 Asp Ala Ile Cys Glu Val Lys Pro Cys Ser Phe Asn Gly Val Tyr Gln 530 535 540 Pro Ser Leu Leu Asp Thr Phe Pro Arg Gly Gln Leu Leu Ala Leu Ser 545 550 555 560 Tyr Phe Thr Asp Arg Ile Lys Pro Leu Leu Pro Ser Ser Ser Ser Ser 565 570 575 Thr Leu Ser Ile Ser Glu Leu Thr Ser Met Ala Lys Asp Val Cys Ala 580 585 590 Gly Pro Asp Ala Trp Ala Asp Arg Trp Gly Ser Asp Ala Thr Ala Met 595 600 605 Glu Glu Leu Ala Gly Arg Pro Glu Tyr Cys Leu Asp Leu Thr Phe Met 610 615 620 Asn Ala Leu Leu Gly Leu Gly Tyr Glu Leu Ser Pro Glu Arg Glu Leu 625 630 635 640 Met Val Glu Lys Lys Leu Arg Gly Val Glu Leu Gly Trp Ala Leu Gly 645 650 655 Ala Gly Leu Ala Leu Val Glu Lys Ala Glu Leu Thr Cys Thr Ala 660 665 670 <210> 17 <211> 656 <212> PRT <213> artificial sequence <220> <223> FBP26 >CNAG_04221 Transcript 1 <400> 17 Met Ser Ile Pro Pro Pro Pro Ser Asn Lys Ser Pro Ala Ser Ala 1 5 10 15 Val Ser Pro Ser Lys Pro Arg Ser Pro Lys Leu Lys Pro Leu Thr Pro 20 25 30 Thr Ser Glu Lys Pro Ser Arg Thr Asn Asn Asp Asp Asp Gln Val Tyr 35 40 45 Gln Pro Val Glu Pro His Val Leu Ala Glu Ala Val Ser Lys Leu Asp 50 55 60 Met Ile Arg Ser Ala Pro Ala Pro Met Ser Thr Val Thr Ser Pro Ala 65 70 75 80 Ala Ser Ala Ala Pro Ser Gly Pro Ser Ser Pro Arg Leu Ser Gly Ala 85 90 95 Gly Gln Gly Ala Pro Ser Thr Gly Pro Trp Ala Met Asp Arg Thr Ala 100 105 110 Ser Gly Asp Gly Arg His Ser Ala Pro Gly Thr Pro His Phe Gly Ala 115 120 125 Ser Thr Ala Leu Leu Lys Thr Leu Asp Glu Thr Thr Lys Val Ile Arg 130 135 140 Gln Ser Ser Arg Ala Pro Ser Arg Ala Pro Ser Val Ser Gly Ile Gly 145 150 155 160 Thr Val Val Glu Lys Pro Asp Tyr Ser Glu Ala Lys Ile Val Val Ala 165 170 175 Met Val Gly Leu Pro Ala Arg Gly Lys Ser Tyr Leu Ser Asn Arg Leu 180 185 190 Met Arg Tyr Leu Arg Trp Leu Glu Tyr Asn Val Gln Val Phe Asn Val 195 200 205 Gly Gln Leu Arg Arg Ser Lys Ala Arg Ser Ala Leu Gln Ala Gly Gln 210 215 220 Gly Lys Val Asp His Ser Ala Thr Tyr Phe Ser His Ser Asp Ala Glu 225 230 235 240 Ala Thr Lys Lys Arg Glu Glu Leu Ala Glu Glu Ser Leu Glu Ser Leu 245 250 255 Ile Ser Trp Leu Lys Lys Glu Gly Asn Val Gly Ile Met Asp Ala Thr 260 265 270 Asn Ser Thr Ile Asp Arg Arg Glu Lys Ile Lys Ser Arg Ile Asp Lys 275 280 285 Glu Pro Gly Leu Gln Val Leu Tyr Leu Glu Ser Phe Cys Asp Asp Pro 290 295 300 Val Val Ile Ala Thr Asn Ile Ala Leu Lys Val Arg Ser Gly Asp Pro 305 310 315 320 Asp Tyr Gln Gly Met Ser Lys Glu Asp Ala Glu Arg Asp Phe Arg Lys 325 330 335 Arg Ile Ala Gln Tyr Glu Ser Val Tyr Gln Thr Ile Asn Glu Pro Asn 340 345 350 Ile Pro Phe Cys Arg Ile Leu Asn Val Gly Gln Arg Val Thr Ile Asn 355 360 365 Lys Ile Glu Gly Tyr Leu Gln Ser Arg Ile Ala Phe Tyr Leu Met Asn 370 375 380 Leu His Leu Lys Pro Arg Ser Ile Tyr Leu Ser Arg His Gly Glu Ser 385 390 395 400 Met Tyr Asn Val Glu Gly Lys Ile Gly Gly Asp Ser Asp Leu Ser Pro 405 410 415 Arg Gly Trp Glu Tyr Ala Arg Ala Leu Pro Ala Leu Ile Lys Asp Asn 420 425 430 Ile Gly Glu Gly Pro Leu Glu Val Trp Thr Ser Thr Leu Gln Arg Thr 435 440 445 Gln Gln Thr Ala Ser Tyr Leu Pro Phe Glu Lys Lys Thr Trp Lys Ser 450 455 460 Leu Asp Glu Leu Asp Ala Gly Val Cys Asp Gly Met Thr Tyr Lys Glu 465 470 475 480 Ile Glu Gln Lys Tyr Pro Glu Asp Tyr Glu Ser Arg Asp Asp Asp Lys 485 490 495 Phe Asn Tyr Arg Tyr Arg Gly Gly Glu Ser Tyr Arg Asp Val Val Val 500 505 510 Arg Leu Glu Pro Val Ile Met Glu Leu Glu Arg Gln Asn Asn Ile Leu 515 520 525 Ile Ile Ala His Gln Ala Ile Leu Arg Cys Leu Tyr Ala Tyr Phe Gln 530 535 540 Ala Arg Pro Gln Gln Glu Leu Pro Tyr Ile Asn Ile Pro Leu His Thr 545 550 555 560 Leu Ile Lys Ile Thr Pro Gln Ala Tyr Gly Cys Gln Glu Glu Arg Tyr 565 570 575 Pro Leu Pro Ile Ala Ala Val Asp Thr His Arg Pro Arg Pro Ser Lys 580 585 590 Gly Arg Asn Thr Ala Gly Val Ser Val Ala Glu Glu Ala Phe Gln Pro 595 600 605 Val Lys Arg Asp Tyr Tyr Gly Asp Ser Gln Gln Gly Val Gly Phe Gly 610 615 620 Leu Lys Pro Glu Ala Ile Ser Gln Ala Leu Glu Asn Glu Met Glu Gln 625 630 635 640 Gly Lys Leu Thr Pro Arg Ala Ala Val Ala Ala Gln Leu His His Glu 645 650 655 <210> 18 <211> 613 <212> PRT <213> artificial sequence <220> <223> PSR1 >CNAG_04224 Transcript 1 <400> 18 Met Pro Thr Thr Arg Thr Glu Pro Pro Thr Val Ala Pro Ala Ile Ser 1 5 10 15 Pro Gln Asn Thr Asn Thr Ala Gly Pro Ala His Thr Thr Ser Ile Asp 20 25 30 His Asn Thr Ser Thr Thr Asp Thr Gln Gln Pro Ser Ser Gly Leu Gln 35 40 45 Pro Ser Ile Leu Pro Pro Val Ala Thr Pro Ala Thr Gln Asn Leu Ala 50 55 60 Ser Thr Thr Glu Met Thr Lys Asp Gly Gly Ala Ala Ala Ala Gln Pro 65 70 75 80 Ser Thr Ala Gln Thr Thr Leu Pro Glu Pro Gly Thr Thr Ser Thr Ser 85 90 95 Ile Lys Pro Thr Glu Gly Glu Gln Ser Lys Gly Thr Pro Leu Gly Asn 100 105 110 Leu Ser Arg Arg Leu Ser Asn Lys Ser Pro Ser Thr Thr Ala Ser Ser 115 120 125 Ala Pro Gln Thr Thr Ala Glu Lys Ala Asp Pro Lys Pro Ala Ser Ser 130 135 140 His Thr Gln Pro Thr Thr Ser Thr Ser Lys Thr Thr Val Asn Thr Pro 145 150 155 160 Ala Ser Arg Ser Val Asn Gly Ala Thr Lys Ser Lys Thr Ala Pro Thr 165 170 175 Ser Asn Thr Thr Ala Pro Lys Ala Gly Gln Lys Lys Lys Arg Lys Arg 180 185 190 Lys Gly Leu Ala Gly Ile Leu Leu Ala Leu Gly Cys Leu Ser Val Asp 195 200 205 Glu Phe Glu Glu Glu Pro Ser Lys Pro Ser Ser Thr Thr Ala Ser Val 210 215 220 Gly Ala Gly Lys Thr Ala Gly Ala Gly Ala Thr Thr Gly Val Ser Thr 225 230 235 240 Lys Ala Asp Glu Ser Ala Lys Pro Gly Ser Gly Asp Ala Gly Met Thr 245 250 255 Ser Gly Ala Leu Lys Ala Pro Asn Gly Ser Val Ala Pro Ala Pro Ser 260 265 270 Gly Pro Ser Ala Val Lys Thr Gln Asp Thr Thr Val Gly Ala Glu Gln 275 280 285 Lys Val Asp Ala Thr Gly Pro Thr Gly Ser Thr Val Val Ala Glu Gly 290 295 300 Ser Asn Glu Ala Asp Lys Gly Ile Val Pro Asp Glu Gln Val Val Val 305 310 315 320 Pro Pro Thr Glu Pro His Thr Leu Pro Asp Asp Glu Thr Ala Gly Val 325 330 335 Thr Ser Ser Ala Val Gln Pro Pro Gly Gly Gly Ser Val Leu Leu Gly 340 345 350 Thr Pro Ser Lys His Val Ser His Arg Glu Ser Glu Thr Asn Leu Gly 355 360 365 Thr Ser Ser Asn Glu Arg Thr Glu Thr Ser Gly Gly Tyr Ser Asp Ile 370 375 380 Ser Asn Ser Glu Met Val Asp Glu Ser Thr Gly Gln Gly Gly Asp Glu 385 390 395 400 Leu Gly Glu Asp Tyr Leu Glu Tyr Asp Asp Glu Glu Asp Arg Leu Ile 405 410 415 Glu Gln Gly Gly Ile Gly Ile Pro Val Asp Glu Asn Gly Asn Pro Ala 420 425 430 Pro Leu Leu Pro Pro Ile Ala Ala Lys His Arg Gly Arg Lys Cys Leu 435 440 445 Val Leu Asp Leu Asp Glu Thr Leu Leu His Ser Ser Phe Lys Gln Leu 450 455 460 Pro Thr Ala Asp Tyr Ile Val Pro Val Glu Ile Glu Ser Gln Val His 465 470 475 480 Asn Val Tyr Val Ile Lys Arg Pro Gly Val Asp His Phe Leu Thr Glu 485 490 495 Met Ala Lys Ile Tyr Glu Ile Val Val Phe Thr Ala Ser Leu Ser Lys 500 505 510 Tyr Ala Asp Pro Val Leu Asp Met Leu Asp Glu Asn Arg Val Val Ala 515 520 525 His Arg Leu Phe Arg Glu Ser Cys Tyr Asn His Lys Gly Asn Tyr Val 530 535 540 Lys Asp Leu Ser Gln Leu Gly Arg Asp Ile Gln His Ser Ile Ile Ile 545 550 555 560 Asp Asn Ser Pro Ala Ser Tyr Ile Phe His Pro Asn Asn Ala Val Pro 565 570 575 Val Ser Thr Trp Phe Ser Asp Pro His Asp Ser Glu Leu Thr Asp Leu 580 585 590 Cys Pro Phe Leu Ala Asp Leu Ala Thr Val Asp Asp Val Arg Gly Val 595 600 605 Leu Asp Gly Arg Ile 610 <210> 19 <211> 706 <212> PRT <213> artificial sequence <220> <223> CDC1 >CNAG_06647 Transcript 1 <400> 19 Met Leu Gly Ser Pro Arg Ala His Ser Pro Ala Pro Pro Leu Ala Ala 1 5 10 15 Arg Gly Arg Lys Thr Gly Leu Lys Ser Arg Ala Thr Gln Ile Leu Ala 20 25 30 Leu Arg Phe Gly Trp Val Val Leu Val Ile Trp Tyr Glu Val Gly Glu 35 40 45 Phe Phe His Ser Leu Ser Thr Cys Arg Phe Pro Asp Ser Ala Leu Arg 50 55 60 Gln Ala His Pro Gln Ala Pro Pro Pro Thr His Val Val Leu Ile Ala 65 70 75 80 Asp Pro His Val Pro His Ala Arg Leu Ser Tyr Pro Ser Gly Asn Pro 85 90 95 Trp Leu Asn Trp Ala Lys Gln Gln Met Asp Glu Leu Phe Met Arg Lys 100 105 110 Ser Trp Asn Val Val Met Arg Leu Gly Arg Val Asp Gln Val Leu Val 115 120 125 Leu Gly Asp Met Leu Asp Ser Gly Arg Gly Val Met Ser Asp Glu Glu 130 135 140 Tyr Val Glu Tyr Ile Ala Leu Phe Arg Ser Ile Phe Gln Leu Pro Pro 145 150 155 160 Thr Thr Pro Met His Phe Val Pro Gly Asn His Asp Ile Ser Leu Val 165 170 175 Pro Asn Gly Arg Phe Ser Ser Gln Ala Arg Leu Arg Tyr Gln Gln His 180 185 190 Phe Lys Thr Pro Asn Thr Val Leu Pro Ile Ser Asn His Ser Phe Ile 195 200 205 Leu Leu Asp Ala Val Gly Leu Val Glu Glu Asp Tyr Arg Arg Tyr Ala 210 215 220 Ser Glu Met Gln Phe Gly Glu Trp Asp Gly Val Lys Gly Gly Val Ile 225 230 235 240 Glu Phe Val Lys Asp Leu Arg Asp Asn Pro Pro Pro Gly Pro Lys Ile 245 250 255 Leu Leu Ser His Ile Pro Leu Ala Arg Pro Glu Gly Ala Ala Cys Gly 260 265 270 Pro Leu Arg Glu Lys Gly Arg Ile Ser Lys Gly Ala Gly Pro Gly Tyr 275 280 285 Gln Asn Leu Leu Gly Ser Glu Thr Ser Lys Phe Leu Leu Asp Ala Ile 290 295 300 Gln Pro Asn Ile Val Phe Ser Gly Asp Asp His Asp Tyr Cys Asp Tyr 305 310 315 320 Val His Lys Gly Asn Ile Arg Glu Val Thr Val Lys Ser Phe Ser Ser 325 330 335 Ser Thr Gly Ile Arg Arg Pro Gly Leu Gln Leu Leu Ser Leu Val Pro 340 345 350 Pro Pro Thr Glu Ser Thr Ala Arg Leu Leu Pro Thr His Ala Asp Arg 355 360 365 Pro Cys Phe Leu Pro Asp Gln Leu Gly Val Tyr Trp Arg Val Tyr Leu 370 375 380 Pro Leu Ala Ile Leu Thr Ala Leu Tyr Leu Phe Ile Thr Asn Leu Arg 385 390 395 400 Ser Ala Tyr Leu Arg Trp Asp Arg Ser Ser His Ala Val Ser Glu Lys 405 410 415 Met Arg Ser Ser Pro Ala Leu Leu Ser Ala Glu Thr Met Ser Pro Asn 420 425 430 Ser Phe Ser Ser Arg Arg Asn Gly Pro Val Pro Leu Ser Ile Pro Ser 435 440 445 Arg Lys Ser Ser Ser His Leu Pro Leu Ser Ala Pro Ser Ala Ile Pro 450 455 460 Ser Ser Thr Leu Pro Arg Pro Val Arg Tyr Asn Ser Thr Pro Ala Glu 465 470 475 480 Tyr Pro Pro Gly Ser Arg Ser Gly Gln Ser Asn Pro Val Ser Pro Phe 485 490 495 Gly Ser Pro Lys Leu Ser Ala Val Glu Arg Phe Gly Glu Arg Asp Val 500 505 510 Glu Arg Asp Gly Glu Ala Ala Ser Ala Ser Val Thr Gly Leu Asn Thr 515 520 525 Pro Leu Thr Leu Ser Arg Arg Ser Ser Tyr Ile Tyr Met Asp Arg Gly 530 535 540 Phe Pro Ser Ser Val Ser Asp Ser Ala Pro Leu Ser Ala Ser Gly Thr 545 550 555 560 Thr Asn Trp Gly Leu Gly Ala Asn Thr Gly Val Ser Ser Pro Ser Ser 565 570 575 Ser Gly Phe Ile Arg Arg Val Ser Ser Ser Ala Asn Leu Ser Thr Leu Ile 580 585 590 Thr Thr Asn Val Ala Pro Pro Ser Leu Ser Ile Thr Ser Pro Gly Thr 595 600 605 Pro Arg Arg Val Thr Leu Pro Ser Pro Leu Leu Leu Pro His Ser Pro 610 615 620 Ala His Ala Gln Ala His Pro Leu Ser Gln Thr Ser Ser His Ala Thr 625 630 635 640 His Pro His Pro Ala Val Ile Tyr Thr Phe Pro Thr Pro Ser Arg Ser 645 650 655 Trp Phe Trp Phe Glu Arg Ala Lys Ser Phe Leu Arg Trp Ala Trp Lys 660 665 670 Ala Arg Lys Gly Ala Val Gly Lys Ser Trp Arg Glu Leu Ile Ser Val 675 680 685 Ala Trp Val Gly Ala Ile Val Trp Leu Gly Val Asn Ala Leu Phe Phe 690 695 700 Leu Glu 705 <210> 20 <211> 444 <212> PRT <213> artificial sequence <220> <223> INP5202 >CNAG_07317 Transcript 1 <400> 20 Met Ala Pro Leu Asp Val Phe Met Thr Thr Trp Asn Thr Gly Leu Gln 1 5 10 15 Gly Ser Lys Ala Gln Ser Gln Asp Leu Thr Ser Trp Leu Leu Pro Val 20 25 30 Leu Arg Asn Ala Ser Asn Pro Glu Leu Pro Gln Gly Phe Ile Pro Asp 35 40 45 Leu Tyr Ala Ile Gly Ile Gln Glu Leu Leu Pro Leu His Leu Ala Met 50 55 60 Ala Gly Leu Thr Glu Pro Val Leu Leu Ala Leu Thr Ser Arg Ile Glu 65 70 75 80 Asn Leu Leu Ser Ala His Ala Ser Ser Ile Ser Pro Asn Lys Thr Pro 85 90 95 Glu Arg Tyr Ser Leu Val Ala Arg Val Ala His Val Gly Asn Ala Leu 100 105 110 Trp Ile Phe Ser Arg Asp Ser Thr Met Asp Gly Arg Leu Gly Lys Pro 115 120 125 Ser Thr Ala Thr Leu Gly Leu Tyr Trp Gly Gly Met Gly Asn Lys Gly 130 135 140 Ala Val Gly Val Arg Leu Pro Val Arg Arg Gly Lys Ile Gly Gly Trp 145 150 155 160 Glu Asn Leu Thr Phe Val Asn Ala His Leu Glu Ala His Asp His Asn 165 170 175 Ile Pro Arg Arg Asn Ala Gln Tyr Gln Arg Ile Leu Ser Ser Leu Val 180 185 190 Phe Asn Ser Thr Asp Pro Leu Thr Thr Ser Gln Gln Ile Phe Asp Thr 195 200 205 Ser His Leu Phe Phe Met Gly Asp Leu Asn Tyr Arg Leu Ser Lys Gln 210 215 220 Pro Pro Pro Gly Ala Leu Gln Glu Asn Lys Met Phe Gly Asp Val Leu 225 230 235 240 Glu Leu Glu Lys Ser Arg Met Gly Met Leu Asp Thr Asp Thr Leu Arg 245 250 255 Gln Glu Gln Arg Glu Gly Arg Val Phe Gly Gly Leu Arg Glu Gly Asp 260 265 270 Leu Thr Arg Phe Ala Pro Thr Tyr Lys Arg Ile Val Gly Gln Ile Glu 275 280 285 Gly Tyr Ser Lys Lys Arg Ile Pro Gly Trp Thr Asp Arg Ile Leu Phe 290 295 300 Ala Ser His Thr Asp Pro Pro His Leu Phe Ser Pro Glu Ala Ser Leu 305 310 315 320 Asp Pro Val Pro Ser Asn Val Ala Asp Thr Thr Ser Ile Leu His Phe 325 330 335 Asn Ser Thr Ile Glu Leu Val Ile Ser Asp His Lys Pro Val His Ala 340 345 350 Ile Leu Ser Leu Pro Glu Val Ser His Glu Ala Pro Ser Pro His Leu 355 360 365 Ala Pro Thr Leu Pro Pro Ala Pro Ser Pro His Gln Pro Arg Pro Leu 370 375 380 Pro Thr Gln Arg Glu Val Leu Leu Ile Glu Lys Phe Leu Gly Thr Leu 385 390 395 400 Leu Asp Arg Leu Val Gly Trp Pro Trp Cys Ile Ile Val Leu Leu Gly 405 410 415 Phe Gly Asn Thr Arg Arg Gly Met Gly Val Ser Ala Phe Val Ala Met 420 425 430 Ile Trp Gly Ile Trp Trp Ser Gly Val Tyr Ser Gly 435 440 <210> 21 <211> 976 <212> PRT <213> artificial sequence <220> <223> YND1 >CNAG_07692 Transcript 1 <400> 21 Met Ala Pro Ser Val Thr Pro Leu Thr Thr His Tyr Ala Leu Val Ile 1 5 10 15 Asp Ala Gly Ser Ser Gly Ser Arg Leu Gln Ile Tyr Ser Trp Arg Asp 20 25 30 Pro Asp Leu Glu Arg Ala Glu Ile Leu Gln Asp Val Gln Asn Ile Glu 35 40 45 Arg Gln Gly Ser Ser Ser Ser Ser Ser Lys Glu Gly Ala Arg Trp Trp Trp 50 55 60 Ser Gly Glu Asp Gly Trp Lys Gly Lys Gly Lys Gly Lys Ala Lys Glu 65 70 75 80 Met Glu Glu Met Ala Leu Arg Arg Leu Val Arg Val Gly Lys Gly Val 85 90 95 Glu Gly Asp Asp Trp Val Lys Arg Val Glu Pro Gly Ile Ser Thr Val 100 105 110 Asp Pro Glu Asn Ile Pro Glu Tyr Leu Ala Pro Leu Leu Thr His Ala 115 120 125 Leu Gln His Ile Pro Pro Ser Val His Ser Ser Thr Pro Ile Tyr Leu 130 135 140 Leu Ala Thr Ala Gly Met Arg Leu Leu Pro Ser Gln Gln Arg Asp Ala 145 150 155 160 Ile Leu Gln Ala Thr Cys Asp Thr Leu Arg Asn Asp Tyr Pro Phe Leu 165 170 175 Val Ser Gly Pro Thr Glu Glu Gly Pro Cys Gly Glu Asn Val Arg Val 180 185 190 Ile Asp Gly Glu Glu Glu Gly Ile Trp Gly Trp Val Ala Val Asn Tyr 195 200 205 Leu Met Asp Gly Phe Gly His Ala Pro Ser Pro Ser Ser Ile Ser Asn 210 215 220 Ser Gly Thr Ser Ser Ser Ser Ser Ser Thr Asn Leu Leu Pro Leu Ala Pro 225 230 235 240 Leu Ala Ser Ala Pro Pro Asp Ser Ser Ser Ser Ser Ile Thr Pro Val 245 250 255 Asp Ile Ala His His Ser Pro Thr Phe Gly Phe Leu Asp Met Gly Gly 260 265 270 Ala Ser Thr Gln Leu Ala Phe Ser Pro Ser Ala Ser Glu Leu Leu Thr 275 280 285 Ser Gly Phe Pro Leu Asp Lys Leu Arg Thr Val Ser Leu Arg Leu Leu 290 295 300 Ser Gly Glu Gln Val Asp Trp Pro Val Phe Val Ala Ser Trp Leu Gly 305 310 315 320 Phe Gly Thr Asn Arg Ala Arg Glu Arg Tyr Met Thr Ser Leu Tyr Gln 325 330 335 Gln Trp Ala Ser Ala His Pro Ser Pro Ser Ala Gln Asp Leu Ala Thr 340 345 350 Pro Ile Pro Asp Pro Cys Leu Pro Lys Asp Leu Ser Ile Leu Pro Pro 355 360 365 Ser Ser Ser Gln Pro Pro Leu Ile Gly Thr Gly Ser Phe Pro Glu Cys 370 375 380 Leu Thr Ser Leu His Pro Leu Leu Glu His Ser Thr Pro Cys Pro Thr 385 390 395 400 Ser His Cys Leu Phe Gly Gly Gln Pro Thr Pro His Ile Asp Phe Glu 405 410 415 Arg His Asp Gln Arg Gly Phe Ile Gly Ile Ser Glu Tyr Trp Tyr Thr 420 425 430 Met Gln His Val Leu Gly Val Gly Gly Val Trp Asp Trp Gly Glu Trp 435 440 445 Glu Lys Gly Met Lys Glu Phe Cys Gly Lys Asp Trp Glu Val Ile Lys 450 455 460 Ser Glu Val Glu Asn Gly Asp Trp Glu Asp Val Asn Met Asp Pro Thr 465 470 475 480 Arg Leu Glu Met Gln Cys Phe Lys Gly Ala Trp Ile Ser Asn Val Leu 485 490 495 His Glu Gly Ile Gly Ile Pro Arg Leu Val Asp Val Gly Gly Asn Asp 500 505 510 Thr Leu Thr Gly Gly Ser Leu Gly Asp Thr Asn Ala Glu Ala Glu Arg 515 520 525 Arg Ala Arg Glu Lys Gly Leu Phe Glu Lys Lys Gly Gln Gly Gln Gly 530 535 540 Lys His His Phe Gln Ser Met Asp Gln Val Gly Glu Thr Ala Ile Ser 545 550 555 560 Trp Thr Leu Gly Lys Val Val Ile Glu Ala Ser Lys Ala Val Gln Pro 565 570 575 Arg Ser Gln Glu Met Glu Gly Trp Trp Met Arg His Leu Asn Leu Gly 580 585 590 Ser Met Arg Leu Pro Leu Ser Leu Pro Ile Pro Lys His Leu Glu Gly 595 600 605 Lys Leu Glu Asp Leu Gly Leu Ser Val Val Trp Ile Tyr Ala Val Val 610 615 620 Gly Phe Phe Leu Val Gly Met Leu Phe Ser Arg Ser Asn Arg Arg Arg 625 630 635 640 Gly Val Gly Ser Leu Gly Ser Gly Met Gly Arg Arg Arg Lys Pro Ser 645 650 655 Leu Ser Ser Pro Pro Leu Pro Ala Arg Pro Trp Phe Thr Phe Pro Ser 660 665 670 Phe Phe Ser Gly Pro Ala Ala Asp Pro Ser Leu Ser Ile Glu Asp Gly 675 680 685 Pro Asp Ala Ser Pro Thr Ser Ser Thr Ser Ser Thr Pro Phe Ser Gly 690 695 700 Asn Gly Thr Ala Gly Gly Ala Ser Gly Lys Ser Arg Ile Val Pro Gly 705 710 715 720 Arg Leu Arg Leu Trp Ser Leu Arg Ile Ser Asn Thr Ile Asn Lys Tyr 725 730 735 Ile Pro Ala Ser Leu Pro Leu Ser Leu Gly Ser Pro Asn Ser Arg Gln 740 745 750 Arg Gly Gly Ala His Glu Leu Trp Thr Ser Ile Gly Ile Gly Leu Pro 755 760 765 Arg Thr Arg His Asn Ser Met Pro Met Ile Gly Met Gly Pro Asn Thr 770 775 780 Ser Pro Arg Val Gly Leu Leu Ser Pro Gly Gly Asp Gly Gly Tyr Ser 785 790 795 800 Gln Pro Gly Ser Pro Arg Ile Ile Ser Ala Pro Phe Phe Ile Pro Ala 805 810 815 Ala Ala Pro Gly Ile Gly Gly Leu Asn Thr Gly Val Gly Ser Leu Thr 820 825 830 Pro Glu Thr Val Leu Thr Gly Ile Ser Ser Ala Thr Ser Val Ser Pro 835 840 845 Ser Pro Ser Leu Ala Ser Thr Ser Ser Pro Pro Pro Pro Arg Ser Ser 850 855 860 Leu Lys Pro Gly Lys Ser Gly Arg Pro Phe Lys Pro Arg Gln Asn Ser 865 870 875 880 Asn Asn Leu His Pro His His Gly Ser His Gly Phe His Ser Val Gly 885 890 895 Glu Gly Ile Gly Ala Gly Gly Gly Gly Trp Asn Asp Pro Pro Leu Ala 900 905 910 Met Leu Ser Ser Pro Gly Ser Gly Thr Gly Pro Ser Gly Ser Gly Ala 915 920 925 Ala Asp Asp Gly Gly Val Leu Thr Pro Thr Ala Asn Gly Gly Leu Ser 930 935 940 Asn Gly Ala Leu Ser Arg Asn Ser Ser Arg Ala Asn Leu Ser Glu Leu 945 950 955 960 Gly Leu Ala Gln Arg Ser Met Ser Arg Thr Gly Thr Pro Gly Phe Asp 965 970 975 <210> 22 <211> 1283 <212> DNA <213> artificial sequence <220> <223> vps29 <400> 22 cagaagcaaa cctcttccgc ttatattact ctgacctttc cctcgaattt gtatcaccat 60 ggtcctggtt ctcgtcattg gagaccttca tatccctaac cttgtccacg atcttcctgc 120 aaagtttaag aagttactgg tccctggaaa gatcggccaa attatatgta ccggaaatgt 180 ctgcgacaag gagacctatg actatttacg gacgacggcc cctgaagtac acgtagtgcg 240 gggagaattc gacgagaacc ctcatttccc tctgtcactt ataattcagc atcaatcact 300 tcgcataggc gtagtccatg gacaacaggt tgtgcctgct ggagaccccg atatgcttgc 360 agctttggca aggcagatgg atgtagacgt tttgatcagt ggagggacac accgctttga 420 gtcatttgaa ttcgaaggtc gtttcttcgt taaccccggg tcggcaaccg gggcctggag 480 tagtctctgg aatggcgagg taacaccatc attcgccttg atggacattc aagggcccgt 540 cattgtcact tatgtatatc aacttgtaga cggagaggtc aaggtcgaca aagtcgaata 600 ccgtaaacct gaccttacat cagagactca gtctcaatca acgcggtcag aggttgctgc 660 aagatggtag ataatacact tagaagcgaa gcctttgcca tagataagat agcaggtacc 720 ctactatgtg ctggcagctg tctgtaaatg aagctaacag tcagttttgg ccatgtgaca 780 cttacagtat cttaagagca aaattttgct ctggcatgaa ggttgtccat tcaattgcaa 840 acgaacattc tcgaccacga cagtaaggtc aaattcattt caagcactcg aattaggggg 900 atgcaggacg ctgagatata ctagcctctc ttatctaaac aggtaaagca aaccaaattg 960 taaagcgaag atgaggagtc ttctttgaag gacgcaaagc cattccttat cacatactct 1020 cacctcattt aaacgtcttg ttaaatggtt tctttttttt ccatctgatg acttgtttcc 1080 aacagacgtt cgatcattgc acataattta tccactgatc taactagcta gtttctcagt 1140 gctctaactt ccattaatga cgttaaaaaa tgtcctttcg gggccaaagt aaccactact 1200 taggttagct ccctgaaagt ttgacttagc aacttcaaca actccggcaa cttgcaacct 1260 catgcatttc tctagtcttc ttg 1283 <210> 23 <211> 3261 <212> DNA <213> artificial sequence <220> <223> YMR1 <400> 23 cgaatcccat ctcatctcat ctcttctctt cccctgcttt accttacgca tcgcttgtct 60 caactccagc agctatggac gcattacgag tagcaagagt agataacgta acgatccaat 120 actttcttcc cccaacagct cccgatcaaa agccaactcc cctcactcag atcggccagc 180 tacatctcac cccgcatcat ctcatattct ctcatacacc ctctacagct tacgaaccag 240 aaatatggat accttaccct ttgataaccc gtctcactcg tctaccccag acaatcaacg 300 gtctctatcc ccttcaagta gaaacaaaaa cctttgaaag ctacgtcctt ctcttcacca 360 aggacaggga cgatggtgca gaggaagttt ggcagagtgt caaagattgc tctgtcaaat 420 catctgtcga acagctgtac gcattcttct atgtcccccc atcacccggt acaggctgga 480 ccgtcttcaa ccatcgaacc gagtttgccc gacagggttt aggcactcga accaaagcct 540 ggcgattcac agacataaac aaggactact ctttttctcc gacatatcca agcaagctcg 600 ttgtaccgag tcgtatcagt gattctacgc tcatgtacgc gggtaagtac aggagcaagg 660 cacgtatacc agcattgact tacctccact gggccaacaa tgcttctatc acacgatcat 720 cacaaccaat ggttggtata aaaaactcac gctcatctca agatgaaaga ctggtcgagt 780 gtatattttc atcccacatg ttccttgaca atgcctattc ctctgccccc atcttcggag 840 ccacctctac caatctcatt atagacgctc gcccaaccac gaatgccatg gccaatgtag 900 caatgggcgc gggaacggag aacatggaga attataaact aggcaaaaag gcctatctcg 960 ggattgacaa cattcatgtc atgcgaaata gccttaaaac ggtcgccgaa gcaattagag 1020 aggctaactt gagaccatcg gtccccctga atcgagctct tttacgcaag agtaactggt 1080 tacgacatat ctcgacgatt ctcgacggtg ctctcatcat tgtccgcaat atacacctca 1140 acgcctcaca tgttcttatc cattgctctg atggatggga ccggacaggc caactaagcg 1200 ctgtcgcaca gatatgcctg gacccttact accgtacgtt tgacggattc aaggtgctcg 1260 tagagaagga ctggttagca tttggccaca aattcctaga ccgctctgga catttgtctt 1320 cggaaaaata tttcatggtg acagagaatg acgacgacat ggaggaggag ggggtgagtg 1380 cgcaacgtgc cgcgcaagca ttcttcgcga cagtacagaa gcaattcact agcacctctc 1440 acctcaagga gatttcgccg gttttccacc agttcctcga ctgtgtacga cagatccagc 1500 gccagtttcc cgaacgtttt gagttcaacg aacagtacct tttagatata tatcgacatc 1560 tttacacttg ccagtttggt actttcttgt tcaacaatga gcgggaacgt caggaaagtg 1620 cttccccatc acgcaaatca tttgtagagc aaacatgttc tgtatgggac tatctcgact 1680 caccttctga gcgtgaaaag tatatcaact ctttgtatga taccacactt gacagcaatc 1740 aatcgcggga tgcgggggcc gatcaaggtg tgctgtttta taatcccaaa gatgttaggt 1800 tttggttcag gctctttggg cggggagatg aagagatgaa tggatcatct ctgaccttaa 1860 atcagcctca ggggggttgac atcattggtc ctattggggg agatcaagtg gaagatatgg 1920 ctgcaggaga gatactgcga ggggcgtctc cagtttcagc tccgtctcct catgccacag 1980 ccagccagag tcgctcttgg aactggtctc aactttctgg gaatgccctt aatgctgttc 2040 attcagcagc cagagagatc aagagcatat cgcaagacgc attgtctcag attagggccg 2100 aggcaaatga gttggataga gaatcttggg aacaagacgg caagggaaag aacagtgagc 2160 cggcatccct cacggaatct actttgcttc cagagaccaa cccatggtct gctgaggctc 2220 gttcatctcc gaccatacca cctcctcggc cgaacactca ggtgtcgcgg acgactcaaa 2280 acccatgggc agccatgcca gacacaatta cttcgttatc taatttaacg cttgacggca 2340 aggccccggg ttccccagca aacgatgctg gaacgaagga gagagcagga gaaaagcaac 2400 aaaaggcttg ggatcctcta ggagcattat aacgaataca tataccatta tagggttaga 2460 gttccctgac agatacttct ggttgtacat ccacatctag tccttttgaa agtctcttcc 2520 aacagccttc aagggccttt tctccctcag cgtctaaaca tagctccaaa gcgaccgcac 2580 gcgtatctaa agcctgccct gctggctcat cgtcgtcaga tttgacacct tggccttttg 2640 ttacaccttg ctcgtcagag aatatcgggc tttggaaagc tgctttgcgc cactctactg 2700 cacggtcaac cagtcgctga aagtaactgg ttgcaatcaa atgttcctcc cgttcctctt 2760 ggatggcttt cagagcttta gatagacgat atacaatggg ataatgaatg ccaaaaacgg 2820 acattgacat gacgagtggt ttcaccgcgt ctgagcaaag ttgtcagttc cttgactttt 2880 tttttttttt tcgtttttcg cgtaccttga tacagccctg cggacatata tgcatcagca 2940 gcgctggcgg cgatgccggc aaactggcct ttccactgca agagtcagca tatttgaaca 3000 tggctagcat acttggcaac tcactatgct tttactgcta tcagcaagac ttttggcgga 3060 ggctttgtct ccgcatttta cggctgattt caagccaata tctagtagct ctctagaaag 3120 tccacctaga gactttgctt tcggagatgc accaaacttc aggagaacaa gctcaagggc 3180 gccttggtaa tctggcgggt cggcgaaata aaggctgctg gccatatact ctgtttcgtg 3240 agcgttgtac ttgctggttt c 3261 <210> 24 <211> 1313 <212> DNA <213> artificial sequence <220> <223> SSU72 <400> 24 cgagttttct tcagttgatc tatgggacgt atcaccgtgc ttgcctattc ctcatctcgt 60 tcgcccgcaa aacaacatcc gctagccaat ctgtccagcc agccatggac ccccgccgtc 120 gccacaacca gcgtccccca ccgccgtcat cctcattgcc tcccaaccca gctgcctata 180 atgctcctcc aaactcgtat ggaggatcat atcctgatgc acggcaatac cagggccata 240 atggcgcgca tagtaccccg caaggataca gaagtgctcc tccgccgcaa ccgccatatg 300 gtgctttacc tggcgaacaa agagcctttc cacccagcaa catgccaaac tatcctccca 360 gtggccctcc agacccccgc atgaggcctt ctcaagatcc cagatctaga ttatcgggtt 420 ctcaagggaa ttacaatact ccaaccccac cttcggggca caccccctcct tctttaccca 480 attatggcac cccacctatc tctgcgccca caataccttt accgtcgcaa cagagtcacc 540 agcaattcta tacgcctcca agcggcccta cgtcttcact tcctggtgcc atgccttcag 600 gtgtatttc ggaaccagca aatggatttg tggacaagga tgtgccacaa ggaagaagaa 660 gaccgttatt ttgtgtggtt tgtgcgagta ataacaatcg atccatggaa gcacactatg 720 tcctgaacaa aaattccttc cgagtcgtct ccgccggaac aggttctgct gtccgtttgc 780 cgggtccggc catcgacaag cccaacgtct atcgctttgg taccccgtac gatgacatat 840 acagagatct cgagtctcag gaccctcagt tatatactcg taatggtatc ctgcctatgc 900 tggataggaa cagaaaagtc aaaaaagcgc ctgaaaagtg gcaagagctg aagagtgtac 960 tggcagacgt tgtcattaca tgtgaagaaa ggtgttacga cgctgtttgt gatgatctct 1020 tgacaagaag tggagagtat aaccgcccga tccacatcat caacattgag atcaaagata 1080 accccgaaga agctcatatt gccggccaat ccatattgga gcttgccaga gccattgaag 1140 cttcagacga ccttgattct gatattgatg ctatcctcaa cgctcacggt gacaagcacc 1200 ctcacacgct tttgcatacc gtaggattct attagatata acactataac actattaaaa 1260 ggaatgcaaa agggttaaga tgcatataaa ggatggatac ttatatacta gca 1313 <210> 25 <211> 1670 <212> DNA <213> artificial sequence <220> <223> NEM1 <400> 25 ccgcccggcg gggatttaac tctttctcgc gttggacatt tccaaagagt ctcacgctgc 60 tcagctgcga tgcccttata gccacaccac cagcactcat gaacaccctg agtcgcatag 120 actcctactt ctccgccatc gcttcccggc caacaaccca cccaccacgc accccgccac 180 gtcgctctcg ccagacaatc tcctccatca gcgtcccacc acccacagca ccactcatcc 240 tccgcatagc gctcgtccta tggagcgtcc tcctcactgt ctggaggagc tttgtcggag 300 agacacgtgc gacccgccgc aggggacgcc gctcccgtcg taagcgtctt gccgggctca 360 gagagctcgg cgagcgggtt atgatcactg ccgggatagc gtcgctcgat actccgcagg 420 agcacacaga gggagacgaa ggcagtgagg atgacaagga agacgggtgg gtcgatcctg 480 tcaccagggg accagaaggc tcggcaagtt tggaggaagc gccgcccggg gaagacgaat 540 tcgtgtcggc taacacagcc tcgactggta ccggtgcagc agaggtagag gaggaacctg 600 aacctgaccc tgatgagatg acagtgacag cgaaagacga cagacttgga ggtcctgatc 660 ccaactttac ctttcgcctg cgctcggcac ccaagaagga gctggacggt acagagactg 720 ctgtccattc acccggccac aagccgattc cttcattcca gcggccaccg tctcccacct 780 ctatactcaa caaccccatt accccgtctc cgccgccgcc gccgccgtcg aaaactgtag 840 aaccatcgcc caaacgtccc tctggcaccc gtcttctcgc gaaccccata tcaacatctc 900 tccttgaccc gtctgtcccc gcgccagcgt ccaacgccga ttcgtcgttg ttccggaaac 960 catcccctag gccattacgc cagcccacaa ctccgttcca tcttcaaaag acactgatcc 1020 ttgatctaga cgagacgctc atacactcta cgagccgacc aatccattac cctggtggta 1080 gctctggcgg tggtgggctg ctgggtctta gtgtcggggg tgtgtttggc aatggaaggg 1140 ccaaggaggg ccatactgtc gaggtggtgg tgaatggggag gagtacaatg tatcatgtat 1200 ataagcgccc ttacgtagat catttcctta aaaaggtcgc gtcttggtac acacttgtga 1260 tctttaccgc ctccatgcct gagtatgcgg accccgtaat cgattggctc gatggcggcc 1320 gcaatttatt tgccaagaaa ctgtacagag aaaactgcca tgtgcagccc aatggaagct 1380 acatcaaaga cctgactctg gtcgaaaagg atctgagtag ggtgtgtttc atggacaact 1440 cgcccgtcag ctacagctgg aacaaagcaa atgcgctgcc aatagaaggg tggacgtctg 1500 atccgaacga cgaggcgctg ctgcattcga ttccggtact ggacagtctc aggtttgtga 1560 atgatgtgcg gagggtgttg ggcatccggg ggtttagcta gaagctagag ggacagggga 1620 tatatgcatt gtttcctaat agtcgtgtat gcatggacgg tgttttccgg 1670 <210> 26 <211> 2322 <212> DNA <213> artificial sequence <220> <223> YVH1 <400> 26 acccctgctg tcaaattact tttgtaaata cacatataca gctctcggcc cttactctcc 60 tccgccatgc cccagcacga taccgtcccc tccaggccac cttcagatgc agccaagtca 120 gacaacctcg ccatacaaga cctctcacgc gaggtcacct ccacaggcaa catccgcctg 180 gcggcaaaac gtctcgccca atccgcccaa gcagaaaaag tgagaaatct gaaagaacgc 240 gcgagcacac gggctcaagt cgcacagcca tatgccagat gggcagatga tccagaagaa 300 gctgaatatc ttcagtccaa cgtccatgcc caggcagcag cgcatgtaga agaccaggtc 360 cttgtgagcg atgaggagga ggaagaagaa gagaaagagg ccatgggtca catgcaggaa 420 gtcgtggatg gtctttgggt tggcgatttg gtcgctgcga atgatgatga cgagcttgaa 480 aaaaatggca ttaaaaatat cctgtctgca ctcagaccgt ctctgaaatt ctcagacaag 540 tacgcggtct acccattaga gattgacgac tctgcggata ccgatttact ctcccacttg 600 cctagctgtg tggcatggat caaggaaatt ttagatttac gtcaaaaggc agcagagcct 660 tcatcacaga agaatggtac cgaaaacggc gaatcgctga aaaggtcgcc agacattgat 720 accgtagccc agccgggcaa gccgggaggt gttctggtcc attgccaagc tggcatgtcc 780 agatcagcca gtatcgtggc ggcgtatctg atgagccagt atgatctcga ccccatggag 840 gcaatgacga tgatcaggga gaagagaccc gtagtagagc catctgcgac tttctggcat 900 cagctgggat tgttctacac tacagatggc aaggtatcat taaaagatcg atccactaga 960 cagtactata tggagcgaac cactacgcag ttcatcaacg gtgatggaac agcgccctct 1020 atggaaaaga tggccaagta ccctgcatca ccttccccgt ccaatcctcc tacgcccaag 1080 gaccatgccc gtcgcaagat ccgatgcaaa atgtgccgac gccatctggc tgtgcgagag 1140 catatgatgg accacattct cgatcaagcg cctcctgtac ccgcctcccg gccccgtaca 1200 ccttcaggcg catccatatc aagccaaaga gcaagtttta gcagtaacgc tgggatgaga 1260 tttactgatg tcgtgggaga aggcgcgggt tttttgacag aaagggagcg gaggggtagc 1320 caggtcagcg atgtgatcaa ccccttgact ggcttgcctg gtgcattgtc aagaagatcc 1380 agtgcaggcg ccgggtccaa cggtgctgtc agccctacag caacccagac gctttacgaa 1440 cgagacactg tgacgtcgcc cttgtccatc tcccacaatc atcacaacaa caacaacaat 1500 aacaacacta cccatccggc ctcgcgacga ggacccattc tccgtaatca ttccgagcca 1560 gctggaactg taccaccacc ccccgttccc ttacctgctg ctcatagcac tacttctgta 1620 ccagctcctc aagctcccac gacccagcgt gctttacagt ctgcggacca gcttaacatg 1680 aggttaccgc cgcagctttt agcccttcgg atggcgggta tgggtggtgc cgctgccaat 1740 gccggtgcca gcgcgaatgc ttctaaccct ccagtctccc cgggaacaaa tacaccttcc 1800 ccagtcattg aaaaagaacg cagagaccaa tcttcatctt ctatcaatac caatggcggg 1860 gcaggcgctg cagcaagacg attcagttca cttgcaatga ctcccaagga tgagaaggaa 1920 gaaacgaaat tgtacgagag aagggcgagt ggtggggaag gaatgtatgg cccaccacct 1980 atacttgtca acaacaaatg ctcaggttac tttgttgaac ctctgacgtg gatggagccc 2040 gttctctcaa agggacagat cgcgggaaag ctggtctgtc cgaatgagaa atgcggtgtc 2100 aagattggta attttgattg ggccggcgtg caatgtggat gcaaggaatg ggtgacacct 2160 ggattttgta tccaccgaag caaggtggat gaggttttct aatgtgccag tgagaggggag 2220 taagggactt gggactagtt gtttgcattg catgtagagt tctttgttac gcatagcatg 2280 ttctgaaaga tgcatgatat gatatgtata tgttaacgcg aa 2322 <210> 27 <211> 1130 <212> DNA <213> artificial sequence <220> <223> OCA101 <400> 27 gtatactgcg cagactacca tggcaaacaa gcccccctcg cagcccctca tccaggtccc 60 cgccctcttc tccatagtcg aaccaggcgt ctaccgctcc gccagtccaa ctccgtcgca 120 agtgcccttc cttgccggcc tcaatctcaa aacgatcatt tccctgaccc cggaacatcc 180 tattaagcct cttctacaat tcgttcgtac agcgggcatt tcatttgtcc atctaggact 240 cacccattgg cgccgcccgg gaacggattg gagacctgtc agatatgaaa taattaaaac 300 cgctctcgag gcgtacatct tggatacaag agcgcatccc gtcctactca tcgatccgtt 360 gggggtacat caaactggct gtcttgtggg agcgttgaga atgatgcaag ggtggaactt 420 tgctagtgct ctcatggagt accgtgctca tgctggaagc aagcaccgct atctcgatga 480 acagtatata gagctattcg attcagattt gataaacctg ccagccccac aatatcggcc 540 ttcatggtgg ctgtcttgcg aggaagctga tccgcaagaa gtcaaagcat tggcatcatc 600 cagtggagga acagggctac tcgcggacac gaatggcaga actcaagcaa ttgtctaaaa 660 gattggccgg gtctgccgcc atatcattgc gaactgggat ccaagctcca catcgccgct 720 cgcttcaccg tgaagcctac attggcttgg gcccaatcct ccgtttctgt gttggctaga 780 gttcatatgt ggggagacct tcaaactcca acatggtctc caactcgtaa tccgagcgca 840 gtcacgagat attattgcca ttcatgctgc aattttcccg tctctcctta cctcccgaaa 900 cttccagtag cctgagccgc actgcccata gcagacatcc tctctccaac cttcttattt 960 acctaaaaaa atgcatttcg tcagcaaaaa cttttccaca tgttatccca acagtatttg 1020 aaaaagaaca atcggcttgc actttgtcac ttggcagaga gacttacttc aaaaaatttc 1080 gcagtgcagc ggtcgatgca tacactttcg cccttcaaaa gatccccttc 1130 <210> 28 <211> 1114 <212> DNA <213> artificial sequence <220> <223> SIT4 <400> 28 gatttgcagg atgtgtacta ctacttgtca gcatatgccg agtgatgcgc tatacctcgc 60 cgacgatgcc catcccaatc tcttcagatc cagaccactg gatacagcat atccgccagt 120 gcaagcacct cccagaacgc caaatgaaac tcctctgtaa ccgagtccgc gatcttttac 180 tagaagagtc caacgtccgc ctcgtccagt ctcccgtgac cgtctgtggt gatattcacg 240 gccagttctg ggacgtcctc gagattttta ggcagggagg cgaggtccct aaaactagct 300 atattttcat gggagatttc gtcgatagag gctattatag cttggaaaca ttatctcttc 360 tactggctta caaggcaagg tacccagata agattacgct tttgagggga aaccacgaaa 420 gtagacagat tacccaggtt tatggcttct acgacgaatg tatgcagaag tacggcaatc 480 cttcggtatg gaaagcttgt tgtaacgtgt tcgatcacct caaccttgcc gctatcatcg 540 actcctcaat cctctgcgtt cacggtggcc tctcgcccga tatccgtact ctcgatcaaa 600 ttcgtaccat ctctcgtgcc caagaagttc cgcacgaagg tgcattctgt gatctgatgt 660 ggtctgaccc tgatgaggtt gagacttggt cgataagccc tagaggtgca gggtggttgt 720 ttggggggaa agtgacttca gagttcaact atatcaacgg tctctcgtta atcgcccgag 780 cacatcaact tgttcaagaa ggttacaagc acatgtttga cgaatccctt gtcaccgtat 840 ggtcagctcc caactactgc tacagatgcg gtaatgcggc gagcatcatg caagtagacg 900 aagatggcag gacgagtttc aaagtgtacg acgcggcaat tgaaaattca acggatcaga 960 agaaccctgc aatgagaaga gtgggtgcac catcatactt cgtttgatac cctagatgta 1020 ttttttttgt tgtatcgact atgcatgcct aaggtaaaaa atgccaatgt ataaaaacac 1080 aaaagaagcc agcaaatcaa tgcaacaact tggg 1114 <210> 29 <211> 1766 <212> DNA <213> artificial sequence <220> <223> GUA1 <400> 29 tttttgtcga gcagcaatta tatcttcttt ccacaatcta caacatccat aaccccatac 60 aaaatggcca cagaggagat tcatagcttg tacgacacca tcctcatctt ggattttgga 120 tcccagtact cccatttgat cactcgacga tgccgagagc tcaacgtgta ctgtgagatg 180 ctgccttgca cgcaaaagat ctccgagttg tcctggaagc ctaagggtat catcctttcc 240 ggctcccctt actctgttta cgctcccgac gctcctcacg tcgaccccga cgtcttcacc 300 ctcggcgttc ccatcctcgg tatctgctat ggtctccagg agatcgcccg tgtccacggc 360 ggcaccgtcg atgctcacac ccacaggggag tacggttacg ccaagatcga ggttgtcaag 420 actggcaaga aggatcagga tgcattgttc gagggtattg agatggaggc ggacggtggc 480 540 atcgcgtcca cccctacttc ccctttcacc tccgtcgccc acgaatccaa gcctatttat 600 ggtgtccaat tccaccccga ggtttctcac tcccccaggg gtaaggaggt cattgctgcg 660 tttgtgaaga atgtctgtgg tgtcagagac ggctggagta tggagagctt tatccccaag 720 gagattgcta ggattaggca aatctgcggt gagaagggtc aggttatcgg tgccgtcagc 780 ggtggtgtcg actccactgt cgccgccaag ttgatgcacg aggccatcgg tgaccgattc 840 cacgctatca tggtcgacaa cggtgtgctc cggaaagacg aggccaagaa ggttcataag 900 atgcttaccg ttgatctcgg cgttaacctc accgttattg acgcttccga actcttcctt 960 gcccgtctta aaggtgtcga ggaccccgag cgtaagcgaa agatcatcgg taacaccttc 1020 attgaggtct ttgaggccga ggctgccaag cttgaggctg ctgctgaaaa agagcttgcc 1080 gagaagggcg gtgaggccaa gggcaaaatc gagtggttgc tccaaggtac cttgtacccc 1140 gacgttatcg aaagtatctc tttcaagggc cccagtgcta ccatcaagac ccatcataac 1200 gtcggtggat tgttggagga catgaagttg aagttgattg agcctcttcg agagctcttt 1260 aaggacgaag tccgtgccct tggtcgtctc cttaacatcc ccgagcatct agtcggccga 1320 caccctttcc ccggtcctgg tctcgctatc cgaattctcg gcgaggtcac tcgcgagcaa 1380 atcgcgatcc tccaacacgc cgacgacatt tacattgagg aaattcgtgc tgctggtttg 1440 tacgatcaaa tctctcaagc ctttgttgcc ctcttgcctg tcaaggctgt tggtgttgct 1500 ggtgacgcga ggacatatga ccaggtcgtt gcggttaggg ccgtctctac agaagacttc 1560 atgactgccg actggttcgt gttccccccg caagtgttga agaggatctc gtctagaatt 1620 accaacgagg tcaagggtgt taacagggtg gtctacgaca ttacttccaa gcctcctggg 1680 actgttgagt ggctttaagt tgttagaata tatttcccat tttaggtttt tggatgcata 1740 gatacagact cgattttttg tctttg 1766 <210> 30 <211> 4285 <212> DNA <213> artificial sequence <220> <223> INP5201 <400> 30 ctcagccgca cctatacttt ttgtgtactt tccatcgtcc tccatcatct ttcgtctgcc 60 cgtctacggc ttccggaggg ctacatccgc cttgaggccc gcatttgagc ggtatacata 120 accaggtgca aggaacactg gacgcacatg ttgtccaggt cgtaagcgat ggcatctccc 180 ctgcacgaga acgacgaaga gcgtccacag tcgatagcag ctctcagaag caagtttgag 240 agcctcgcaa tcgctggtgt gagccctgct cccaccgatg tgccctcagc aaccaacggc 300 catgcgactg ttagctctat ccggaatggc ctcctcagtc cccggccaga aacacctgtc 360 gatggtcaaa aggccaagcc tgtgccacca ccaaaaccag cttcacgtcc cgtaagccct 420 gcgactacat cgccagctcc gcaaccttcg tctctgctcc cgcctcctgc acctcgccaa 480 gctcccagca ggcctaccac tcccaaacca tcttttcaga cccatcactc aacatcttca 540 gtcacttcta tcgtgagcgc agcgtctgat tcacatctca aaccttcaga tacaatggca 600 tctcccccag ccgtaatatc gcctgctgta tcacctgcac caacgccgct tcgcaaatct 660 gccccttctg tccccagcaa accgccatct gtcgctgtca ccccctcagg ttcagatggt 720 gatgaagacg agcctgttat cacgtctgta aaggctcttc gtgaaaaatt cagtggtcaa 780 gctcaggcaa gcgaaattgc acttcgaaag cctgtagatg ttcccaaggc atcagcagta 840 tcggtcgtta aggctgcaac ggtacatgat tctcctgagc cgctgtgtgc cccgtcggct 900 acacccatac ctgcacctat tcccgcacca gtcatacaaa gaacgctaga cggcaagaca 960 tcgcccgtaa tgttatcgcc agcatcagag ggcgaagcac tttcagatac caatgattac 1020 tcttcacacc cgactgctcc tctggctcct ccggctcctc cagcacctat ctcacgtata 1080 tcctctcctg tcccagctcc agctccagcc ccttccggcc ctcctcccat aaacagagca 1140 cacaaacctc ctccacgaac tgcaatcagc cccgcaccca tcttccgtcc cgaatcgaat 1200 gtcatcacac ccaacactac atctccccct ataccaggca acaaacctgt catcccatct 1260 cgcagctcta gtgccccaga agctgcagtc ccccctccgc cgccagagcg ccctcaaccg 1320 cctcagctcc ctgtgcgtcg gccgaccttt tcttcgccag atacgcttga gcctagtacc 1380 gcatctgtca tctccccacc tgcactcgct agtactccat tgttgcatac cattcatgac 1440 gatactgctc ttgctcctac gcctgcgcct gctactgccc ccccaccatt gcccgatcgg 1500 tcgagagcta atacaattaa tcgatccgaa agtgaatcga gcgcaaccac cacaggtcca 1560 cctcctcctc gtctaccagc tcggcacgca gctattcctg tatctgctgg tagcgggtcc 1620 acctcttcca acgcaaatgg aagcggcagt accaccatga acccaccccc tcccccggct 1680 catcccgcat ccccatccaa aactcgtatc aactcgggcg gcccaccccc acctctcctc 1740 cggagtgcta ccgttaaccg aggcagcagc gtcggtagcg ggagcgggag tggtggcggt 1800 ggtggcggtt cccctccacg tcgatccaac accatttctc gcgcagcgcc ttttacccaa 1860 gaaaaatact ctacatcagc tacaagcctt ggcctaggcg aaaaaggagt atattcagac 1920 gaagatgacg agcctgaaga gcctggagcg gttaccaacc tgtccgccca agcgaaaagg 1980 atgttggacg agtttccaga catgacagaa gccaaccgtc gtccccccggt gttcgttccc 2040 gatattcgtg tcaaggagtg ccaccacgtt tcggcttttg ctgtttatgg ccgatacgta 2100 tgcacgggcg cacaccatgt acgagtctac gatacccagc tgtcggatca tgcgattagt 2160 gtagtagatt tgaaagagac ggggttagaa agcaggggaa aggacccgaa agtgacggca 2220 atgtgtttcc gtccgggagc gacggaaagt gaagaaggaa ggtacctctg gtgcggtacg 2280 aaagacgggc atctgtggga acttgatatt tccaccgggg aagtaaccag taccaaggcg 2340 tttgtacata cgtcttccat cagctatatc tggcggcacc ggaagaacat catttcgttg 2400 gatgaagggg ggaaattact cgtgtttgat gtaggcgata tagaagggaa accaccgacc 2460 atggcgagac aattacggat aggcgacaaa tttgggttcg ccaaactcat atgcggaaaa 2520 ctgtggacat caagcggtcc ccttacccga tcgactacat cgtccgctac atccaagggc 2580 cctaccgtcc ggatatacga cccctgtgcg ccggggacga tgcccccgcc taaaacgatt 2640 ttcgcaaccg aatgggctgg cgcggtcacg tcggcgacat acatgccttt acatcacgat 2700 accatctttc tcggccacga gggtggattt gtcagtgtgt gggatgggaa agagttggtg 2760 tgtaagcagg tgctgaagat tagctcgacg gatgtgctcg ctttggaggg cgtaggagag 2820 2880 tggttggcga cgaatatatg gattgggcat cccgataacc ctgtgcaatc gttggtggtt 2940 gacccatact caattcagtc tgcgggcaga tacacatgtt ggtcttttgc ccgggatgcg 3000 ctgcgagcat gggacggtct cctctctgtc gactggatcg acaaacaact cactgcacgt 3060 caatcgtcat tctgcacgtt ccgccccgtc aacgttttga tctgtacatg gaacattgac 3120 tctgctaaac ctacagatct gaatggatcg gtcgccaacg cccatttctt ggaagatgtg 3180 ctgaggtctg tggattcacc ggatatcatc gtgtttggtt tccaggaagt catcccgttg 3240 actgataaaa aatatactgc caaaactctt cttttcggga acaaatccaa agatggtggg 3300 gcagcagcag acagggtatc ccacgcctat cgacactggc tagaaaagct tcagtcagca 3360 gtccagatgg cttccccttc aaactgtcca tatatcaaga tccattcaga gagtcttgta 3420 ggcttgttta cgtgtatctt tgtgaaacag tcagagaaaa tttccttacg ggatctagat 3480 attaccactg tcaagcgagg aataggtggg atatacggga acaagggggc tatcgtctct 3540 cgcctcgtga tggatgatac atccatttgc tttatcaacg tccacctcgc tgctggccag 3600 tctcaaaaag cttcgcgaaa cgcggatctt gcagggatcc tagaagataa agccattttt 3660 cctccagcgg acgagttacc gtttgttcat ggaggggtgtg ggacgggaat tttggatcat 3720 gagatggtgt ttttgaatgg tgatttgaat tatcgcattg atcaacggcg tgagaacgtt 3780 atctcatcta tcgccaatgg cgagctagcc tatcttcttg agcatgatca gctgcgtaaa 3840 gagatgagga cgaaccatgc tttcagactg agaaactttg aagaggcgcc catcacgttt 3900 gcgccgacat acaagtacga cccgggcacg cacgattatg attccagtga gaaaaggcgt 3960 attccagctt ggtgtgatag gattctctac aagaaatcgc cacgagtaca agctcttaat 4020 tatcagcgct atgaacctac tgtctcggat catcgaccgg tctctgcagg gtatacgata 4080 atcctgaaag cgatcgattc gttgaagatg atggacgtga gacgggaagc tactggagaa 4140 tgggcgaagc gagaaaagga gttgctagag aagatgcaag aggtgtttga cggtattgaa 4200 taacacttgt tggtgattcg gtatcatatg tagctctaga tttacccaat ggacgattta 4260 tgcatgcatg ttctatgtaa atgga 4285 <210> 31 <211> 2454 <212> DNA <213> artificial sequence <220> <223> PHS1 <400> 31 cggcgttgtg tgcgttgtgg ttgccgtatc gggtggaggt gagatgcggg atgttcccgt 60 ccgacacaaa agcgtcacta cactctgcat tccaaagcgt ccttgactct gcatccccat 120 ccatacttgc ccgcctctca ttgcagctcg ccgccatgcc ccacgtcgct gcgcccagcc 180 gccaagccct cgagcaggag cgtctccacc gccacgccca gcagcccggc tcgtcgctct 240 cccccgcgcc ccgcctctcc ccgctccggc tctacctcct cggctacaac atcctctccg 300 ccctgctctg gggccacctc ctcgtcctca ccctctcctt ccctcctcgcc ccctcccgcc 360 ccccctggca ccagctcgcg gaccgcctca gcggctcata cgactaccac aacctcggat 420 ggtgcaccaa atggacccag accctcgccg tgctcgaagt cgtccacgcc gccctcggct 480 gggtgcgcag tccgctgggc accgtcgcca gccaggtcgc cagccggctc tggaccgtct 540 ggggcgtcgt cgaggccgcg ccggagatta ctcacggcca ccccctgttc acaaccatgc 600 tcctcgcctg gtccctcacc gaagtcatcc gctactcatt ctacgcgctc tccctcctct 660 ccgtctccgc ccccttcctc aactacctcc gctacacaac attcatcccg ctctacccgc 720 tcggcgcctc gtccgaggcc tttctctcgt tcgcgaccct ccctgctttg gcgcccgtcg 780 tcagcagggc ggtgacgaac gtcatggccc aggcgccgag ggagatcatg aagactaaag 840 tggggagaga agtgctttgg tggtctgcga aacatggagg cggcacggcc ggtgcgcaga 900 aagagtgggg ctggatcgag attgtgcgag cgggactgtt tctcctctgg tggcccgccc 960 tttatgtcct ctacacctac atgctcaaac agcgccgaaa ggttttgggt aaaggcaaga 1020 ccgtaggcgg ggttagcaag gcgcaataaa taaaaaagag tagagccggc gcgtggagca 1080 atcagcgatg cattatcggc tggtactgtc agatacgaat ttgaaaacaa ttgaatcaat 1140 cgattgtgga taacaggcag gatcatgata aaccaatgga ctgagagata agtgaaaaaa 1200 tacttttaact ggaggaaaag agatgcgata ataatatcat gctgtgaaat gacgattgat 1260 atggaaacga gagaaaagcg tcgatgaaag aaataaaacc aacatataaa ttctttcatc 1320 agttcatttc tagacatgac atttgtagag aagatcaata tacaccaaaa gacataacat 1380 gtacaatcac aaaaagcttc cactttcaaa acccccctcc ttgctccgaa agctctagct 1440 gagcatctca cgacatgacg ttttaaacta cattctccac cttgatacca tcttcagcaa 1500 cggcattccc ccgcttccct tcccactcct caccacccgt caccaaacaa ggccattcgc 1560 tcatctgttc acctttcccc agtggtcttt tcagtaccgg gtggggact gcagggacgg 1620 aagcagatat gcgggaggtg ggggggatcg agttttcctt cttcttcttg ctgaaaaaag 1680 gtttgggact tttggttttc gccttgaccc ctagggcggt gtaaagtttg tcggtaattg 1740 acaagggttt tgcaaagcca ccaccctcgt ctttaccatt gtcatcccca ttgatgatat 1800 ggtaaggcgg cgagcttgcg cgaggggtaa gcggcaaaga ctcatccgga ccggaggtgt 1860 tgctagtact cgtcgctgg gaaagatacg agggttgacg agactcttcg tcaaatgctg 1920 gtggtttttc cgcggctgga gatggagttg gacacggaga cgaagaagac gaaggataaa 1980 gcgggattct caacacttga ctcgtcttca aactctcagg catcgccatc cccacccaac 2040 cccattcctc ccccctttcc catccaggcg catcatcaaa cgcctcttta tcctcttcat 2100 cctgcaccgc tccctctcct tcacgtccaa caacgacgcc gtcgtgcaac ttgtatcgag 2160 cgggtgtacc ggtcgctttg agtttacaaa cgaatctgtg acgtcgggtg tggtatcgct 2220 gtgaacggat agggtgggaaa gagtcaagaa gcatttgtga gagttcttcg gtacggtagt 2280 gctggagcgg agggaaagtg cgggaggtgg tgtaagtgca tagtttggta gtgcacatgt 2340 cgtcgatcca tgattttgac gatactcccc tatcctcccc ccctaaagaa ggaggggaag 2400 agcgggccca tggagccgaa gagaaggggg cagaagaggt tttggcggtg agga 2454 <210> 32 <211> 4308 <212> DNA <213> artificial sequence <220> <223> MRE11 <400> 32 cctcttccta acttttaagc aaagaaatac ggttttaagc tcaatgtcgg cccccaatcg 60 cgtacccgac tcacaaccaa gtagcgaaat aggagatgag ccgcctccca gtatcgtcga 120 accagatttg gagaattgtt ttcgtattct catcgctaca gacaaccaca taggatatgc 180 ggagaaagat ccggtccgag gacaagactc tatcaacacc tttcgggaaa tactagagtt 240 ggctagagat cacgatgtcg atttcattct tcttgcaggt gacttgttcc atgaaaacag 300 gccaagccga acatgtatgc accagacaat agctctacta agagagttca ccttgggtga 360 caagccaatt gaatttgaac ttttgagtga tccgatggat ggatctacgc ctggtttctc 420 ttttccggct gtcaactacg aagatccaaa tattaacatc gccattcccg tcttctcaat 480 tcatggtaac catgacgatc cccaaggcac tggtcctgag ggtgcactat gtgcattaga 540 tgttctttcc gtttctggag tccttaacta ctttgggaag tcagaccttg tcgctgatga 600 aagtgctgcc gataacccag agaaaggtat tcatatccga ccagttcttc tgcggaaagg 660 tacaacgcat gtggcgctgt atggttgcgg taacatcaga gatcaaagaa tgtatcagga 720 gctacgggca aataaggtca agatgtttat gccgacagga ggtgatgtac ccgatagcga 780 atggttcaac attcttctcg tccatcaaaa ccgcgtccga catggccccc aaaattacgt 840 ccccgaaaac atgtttgatg attctatgcg acttgtcatt tggggccatg agcacgattg 900 taggatcaca cctgagagcg tcgccgataa aaattatttc ataacgcagc ctggaagttc 960 agtggccacc agtttagcac caggagaagc agtaccaaag catgttgggc ttctgtctat 1020 tcaggggtcc caatttcaac tcgaagaatt acctctcaaa acggtgaggc catttgagtt 1080 ggacgaagtt gtgctgtcgt acgctgcgga gcaaggagct gtagatttga acgatagaga 1140 tagtataact tctttccttc gagaacaggt ggaagctttg attttgcagg ctaaaaaaaa 1200 ctggaaggag aggaacaacg gcagcaccaa gaacatgatg cttcccctca tccgactgaa 1260 ggtcgaaaca acggatgcca aagagatggt caatccggtc aggttcggtc aagagtatgt 1320 caatcgcgtc gccaatcctc gagatatctt gcagtactac cgtaagaaaa agaatgagcg 1380 aaaggtcaag aataatcctg atatgccaaa tatcaatgat gatgagtggg aggaagatcc 1440 tgagtctttg actgccgatg aacgactctc caaacttcgt atggcaacac ttgtcaaaca 1500 atatctccaa gctcagagct tagacgtgct ggtggagaat gggatggaag atgctgtgat 1560 gcgctttgta gataaggatg ataaggatgc tatcaaggac tttgtggccg acactctcag 1620 aatggtggga aggaagatga aggagagaga ggttaaagag gatgacgttg atcttgcgat 1680 ggccgaggca aaggagaagg aatataacag atatgccgac agcaacccgg ttccttctca 1740 aagtgtcaaa gggaaaaata aacagcggga ttcagacgtg gatagtatga tggcaagcga 1800 tgatgacatg gatatggacg agatgccgac tcaacagcga gctccagtga gacgtgcaac 1860 cgcgaatcag ccagttagat cagcgaaggg caagggcaag cagcctttgt ttgaaaacgc 1920 ttcagaagaa gaagaggacg aggaagagga agaggaggaa gaagaagaac ctgcgcctaa 1980 aaaaggtcga ggacgagcag cagcggcttc gaccaagaaa gcacctgcga aaaaaccacc 2040 tgcaagaacg ccagctaagt cgacgacaaa ggcaccagct ggaagacgcc ccgcagtcag 2100 tcagccctcg acagggagag gagtaaccca atcacaatta acgttttcaa ggtctggtac 2160 aggcaaggca gcagcagtgc cgatcgaatt gtcatcagat gaggattaga aatgaatgaa 2220 gagggaatct gcgaaaccag tagttattc ttcaacgtca agtgaatcgg atctgattaa 2280 gatcttgatc actgcaataa ttaagaagat attctttcta tgcttgatga tccctttaact 2340 ttagcctttt actcttacgt cacagtcgtt gtgaaaagtt gtgaaaagta cgtaaagtcg 2400 gcattccatg catccccaaa ccaatacaaa cttttgctat tgcttttact tctgactatc 2460 ttttacacac atgccccata gttactccat aacccccaaa agaaaagaaa aaaggaaaag 2520 ctgaacattt tactccttgg cgacctggag gacgattcgg ccggcaatct tgccttcttc 2580 gagaccctcg tagacgctgt cgtcattaat cagcaagcgg ccttgaatta tggataatag 2640 ctgaacttac gccttaaggt cggcaagagg cttctgctca aagatgacct taacctttcc 2700 atcctcgaca agctggaggg attcaatggc gtcttgtcgg ttaccgacgt aagaaccctg 2760 gattcggatg ctcttgaaga cagtccagaa gacgttggca cccatctcgg cgttggggag 2820 accgacagcg accaaagtac cagagggttt taggtagtcg atagcctgag agtaaccagt 2880 cttgtgggaa gcagtcacaa tagcagcggc tgggccttga cccccagtgg cagccttgac 2940 gtcggcaacg aggtctttgg tggtcttaaa atcaacccaa gcatcggcgc cgagagactt 3000 gaccagcttc tccttggcag ccccagtgtc gatggcaacg accttgagac ccatggcctt 3060 ggcgtactgg acagcaaggt gaccgagacc gccaccagca ccgggaaggg caacccaatc 3120 accgaccttt gtgttggaaa ccttgagagc cttataagat gtgacaccag cacagaggat 3180 agaggcagca ccagcagaat caagggaagg ggggatgggg gtaacgtggt tgacgaaaga 3240 gacgacgtat tcggcaaagg taccatcaac tggttagcca gagagctcgg catggtcaca 3300 gtctggacgg atgatcagct ttggaagcga agtccaaata tacagaagat actcactcat 3360 ctcgaagcct cgtcggcaag cctcacaggt gagacaagag ttggcgagcc acttgatacc 3420 gactcggtca ccgagcttca cgggggaatt gacagtgtta gcaccaatgg caacaatgtg 3480 accgacacct tcattattag tcagcgactt gcttcctata aaaattcgat cggcaaggga 3540 cacttacctt cgtgaccacc aatcaaagga ttcatggggg gaattggcca gtcaccctgc 3600 ttggcgtgca agtcggtgtg gcagacaccg gtgtgggtga ttttgacgag acattggccg 3660 gatttcagtt cgttggcctg aatgaccttg gcctgggtgt cgatctcgat ggcaccgccg 3720 acagaggggga cgacggcagc ggtttgagtc ttagggatgg aaacgccttg aagagtggtc 3780 atcttgtaag ctgtgtctgg agtttgggtc tatgttaagt ctggagaatc taagaggtga 3840 agtaccggag tgtctttgct gcggatgttt gtgaaggaaa gggaaaatca tctggatgga 3900 ggtggccgtt atatagtttg agggtgcgag gaaaagtata aacttggttg gcattcatgt 3960 cgatctggat acatagccgg agaatgagcc ggtgagagat gactatgaag ggcggaagtc 4020 atgtgaggtg cctctccggg gtcttatata ccggtgatac agcaatggga ggtaggccgt 4080 agataacaga tacatactgc ttggctgttg gggaatgaag ctaatctctg atttgattag 4140 cggcttaatc gcagtagttt cccgctgcgc tccggggttt tgggccggtg ttgcccggtt 4200 tcaccaccat ctgacgtaaa cccaaacctc tccaccgttt ccaaattccc catctgtctc 4260 gtaatcaagt gcatacatgt tcgttttaga atgatcgttg ctcgccca 4308 <210> 33 <211> 2543 <212> DNA <213> artificial sequence <220> <223> DBR1 <400> 33 cacgagcaga cacagatcag gacagcgcag tatgaggatc gctatccagg gctgttccca 60 tggcagtctc gcccagatat acgacgttgt caactactac tcctcccaaa caaagaaccc 120 tatagacctc ttgctcctct gtggcgactt ccaagctctg cgatcaaaac atgattatgc 180 ctctcttgcc gtaccagcta aattcaagca gcttgggtca ttccatcagt attactctgg 240 tgaacgtgtc gcacctgtct tgacaatagt gatcggaggc aaccacgagg cgagtaatta 300 tatgtgggag ttgtaccacg gtggatggct agctcctagt atatattacc tcggagcggc 360 tggtagtgtt tatgtgaatg gggtgaggat tgtgggtgcg agtggggattt ataagggatt 420 cgactaccga aagggtcact ttgaaaaggt gccttacaat gataaggagc tgagaagtgt 480 ataccatata cgcgagtacg atgtggaaaa gctcatgcat ttaacaccaa gtcctagcac 540 catcttctta tctcacgact ggcccaccac aatagcgcat catggtaaca agaacgcatt 600 gctcaaacgc aagcccttct ttcgagacga aattgaaaag aacacacttg gttcaccgcc 660 cctcttaaga ctaatgaacc atttccaacc ctcttactgg ttctcagccc atctacacgt 720 caagtttgca gctctgtacg agcatcaagc ccctaatcat ggtccggatg tcgacggtgg 780 cgccccctta ccattactgg caacgtcggc cgtaatagct caggctggtg gtaacccaga 840 tgagattcaa atcgatgaag agatggatgc ggggaacccc gatgagatta ttgttgagga 900 tgagggtgaa gaggttatca ttagaccgag acaggtcaac ccggatgaga ttgcgatgga 960 tgacgatgag tttgacgacc ctgcgcccgc ggtccctcag ccattaccag cgacgaccaa 1020 tagcgccttc aatccagaag aaataatcat atcggaccaa gagttcgatg caccaaccac 1080 agtctctcaa ccccttcaac ctctcccacc taccaaaacg aacgcctcca accctgaaga 1140 aatagccatc tcggacgacg aattcgatga ccctgctcct ttggcgcaat ccctcaccac 1200 aattgatgaa tcgaccgacc tcatcgcgca atcacgttct aacccatccc atccacctgt 1260 tgctggcacc atagcccctc ccacttctga ctctattgca tcacgtgtaa tgcaagaagc 1320 gcgacaagag cagcagaaat gggaactgca cggggggaaa gggatggagg gtgtgaccaa 1380 gtttttggca ttggacaagt gtgggcctgg taaagaccat atgcagttcc ttgagatccc 1440 agacccttcc ccacccccaa tcccagggcc tccaagatta acttacgatc ccgaatggct 1500 agccatatcc cgtgctttcc atccatacct ctcaacctca tatcagccca tccccctccc 1560 atcctctgac atacttgagc agatggtgaa ggatgaagta gcaaggatca aggaggaagg 1620 attgctcgtt cccaccgttc cacaagatgg tgcggtcgag ggacaagaag gattagtatg 1680 ggaaaagggc aaagtggatg tcggaagagt gcaaaggttt tggtggactg caccacctga 1740 aggacatccg ggtggaaatg acactgcgtg gtatacaaac ccgcagacag aggcgttttg 1800 tggcatgttg ggtgttcaga acaagattaa tcctcccgtg aacagataat aaaaatctaa 1860 tgatgtgatg aacatgaccc atgcacgaca cgcccgcagt tcccggcttg accgaaaaag 1920 agatatggag aagatttatg tgtattattc tttgcaacct atgtgccata tatgtattgt 1980 gtcatttctc ctgtaacgag tcggcgaccg tggcatgtta gtccaagctt atggagagga 2040 caggattcac catccatggt taaacgaaat gttgtgaatt ggatggagaa agtgatgaaa 2100 gtagtggcac aggcagcaga cgcggccagg ccctaaatcg actggatcga ctgcggatac 2160 cgatgccaaa tcttaattct tggatcttga cttatctcaa aatcgaagat tcagatggag 2220 aaaatatgga atcttgaaac tcatgtttgc ttgaagggac gggcatgggt tcagactgat 2280 gcaagcagga agcacgaaag atgaaaacgc atcaggatag acagcagaaa aggtggtgat 2340 aaccaaatga tttattgtt catcatatgt aatatcaaag taataagcga acaccgaagt 2400 aaaaataaca gattgtccat acaagtgatt gaagatctat cacttgcgag atttcacaca 2460 agattactaa gaggatacgg tcataactga agggaaaaaa atcttataat ctatagtagg 2520 gtcaagagat gtttaggcaa ggg 2543 <210> 34 <211> 1621 <212> DNA <213> artificial sequence <220> <223> SIW14 <400> 34 gttgatgtta agtcggagat aggttgcaaa ccacaactgt tcagatatac cgtcaacatc 60 tccctttttc ttaactatac tgcttcctgg atcactcatt catcggtata cactcatacc 120 gttctcctga cgacacctgt acccaatttt gcgtagccag gggacgccca tgatcccctt 180 ataggtgggc cgtcaccgag cctgctaaac aaccagacga cggaccgagt atcgacgtgc 240 cgggcgaaaa aacggctatt ttccttgaac tatttccatt ttcctacagg attattccct 300 gccaccaaac catgatgtct tcgtcgccaa caatgtcatc gactcctccg caagtaccat 360 cctttttagc gaatatcctg ttatcccatc tttctccttt agatccatca tcgacctcgc 420 caacaacttt cgaacatcat cgttctccca ctacccaaaa cccctcccat cctcaagctt 480 tgcagactgc agaacctccc ccaaaccctc tgtaccttcc tcctcccgct ttaccgaaag 540 ttgaggaaga tctggttcct ccagagaatt ttgcattggt cagtagtggg gtgtatcggt 600 gtgggttccc gaagaaaaga aatttcaagt ttatggagac tttgaggttg aagaccgtcc 660 tgacattggt attggaagaa tatcctaaag caaatctgga atggtgtcag tcccaagaca 720 tacagttcat gcaattcggt ataccaggaa acaaagaacc tttcgacaac attcccgaag 780 acgtaatttg tgccgctctc gttgccatcc ttgaccggcg gaatcatccc atactcattc 840 attgcaacaa gggcaaacac cgtaccggct gtttgatagg ctgcatccgg cgattacaag 900 catggtctct cacttctata ttcgacgaat atcgacgatt ctctgcgccc aaaagccggg 960 cggtggatca acagtttatc gatctgttcg atatcatgcc tgtttgggaa gctgtatgtc 1020 gacctaaagg tgggggatta gggaatttac cggattgggg aatgttggtc ttgccgaaag 1080 gtgtggtgga agtcggtagg gatggaaagg agaagaagag ggttgagaga gatattttgc 1140 atatgcgggg attataatgc tggagcaatc aaatggtgtt tagaaatcat aaattttatc 1200 catagcgaaa ctgtataatg gtagatagat atgttgtact ttgttctacc cccatatatg 1260 ccttcagaac tgcagaagca tcctaaactc catccatcta ctccaaatcc ttttcatcaa 1320 attcggcttc atagggagtc ctcaaatgtc tcgaaggctc ccaaacctgc agaatgttgt 1380 cctccgaagt gctcgcgatt gtccatggcg agctcgggct ccaagaaata tcgcacactt 1440 tgctggtgtg accaccatgg acaaacaaaa gctcaggagg tccgtcttcg gcatcatcgg 1500 gagtttgttc agcaccaata gcatcgagat cccagatgtg gacgcggcga tcagcagagg 1560 cggatgcaaa gtggacaggc gatgtggagg accaggaaag ttgaaggaca tcgttggtat 1620 g 1621 <210> 35 <211> 2931 <212> DNA <213> artificial sequence <220> <223> SDP102 <400> 35 gttgtctttg ttattgttat tatttttgtt attgttgttt tcaaagttct tctttcaccg 60 ccgctatctg ctcctataac aacttcgcag cacatgctca ccccacaatg cagccccagc 120 aacacaccca caacaaccca tcacccgccg cccgaccaca gcctcttcgt gtcgtccact 180 cacccaccat cccacctcca aatcgaggca caagaacttc agagttagct caaggcccaa 240 aagccccatt ggcagcgcca ttaccgttgc gcattactgc ggcgacgtct ccaaatcgga 300 aacgaccgac tccgcttgtg cttgggaagc ccagagaagc tggtccggaa gctggtccgg 360 aagactggga aatacatcag gaaatatctt ttgcggcttc tttgggtgca agtgccgacc 420 attccttgga caatgaattg caagatttat ccaagctacg caaagcagtg cgacaaaacc 480 tcctggctcg accgattgac tctccattag aactgtccgg ttctgaccag tcggccttca 540 atacacccgg acaacaatca tccttcggct cttcctctat atcaatggat agcataccca 600 ttgaacaagt cttcgatcgg gtggagggcg gaagtgtgtt gttggtcgac acccggccgc 660 tggcttcctt tctcaattcc catttaccaa actccatccc actctcggtc cctactctct 720 tatcaaaacg cttccaaaaa tcgcagtctc aatcaagtcc ttcctctata tcatgggcga 780 ctctttcacc ttttgtctct ctatcaagcg ctcgagaacg gtgggactcg gtagatcaag 840 ataaagtcga gattgctgtg atttgtcaag gcgaagaggg tagagttgtg aaggagatct 900 tgaagagctt gatcgaaggc agagttaaag tggtaaaagg tggatgggct gcagtactaa 960 attatgaaag agccagaaga acgctcgttt ccgggcagac cgctacccgc ccctgcctcg 1020 acgtgacttc gccagaaacc gacagcaaac ctcttccccc tgcatctgca tccaatatac 1080 tccctccaaa atcagctccg ccatgtgata tacctctacc acctatccct gcatccccat 1140 ccccaccaaa atctctcaac caccgtcctt cattaccatc acttcgtcca ccttttacag 1200 ggcctactcg gaatcttcct tcactctcga ttaatgccgg tcaagcgagt cagagacgga 1260 cgccaaaatt gagtttgaat tttgacagac ctttgaagag tgctacgctt ggtggctacc 1320 atgatattcc tcccacaccc catgggtttt catgtacgcg aaccaggcca cagaggtctc 1380 ccggattgtc gttaaacata ccccatactc ctttccagcc gcaacaaggt cagattcaag 1440 accggatatt agaagactcc agacccaacg gatccggttc aatacaaacc aaggcgcacg 1500 aacaatcgcg cttcccgcct tcctcttcga catttggcga tgccaagcag attgagaatg 1560 agggggagga catggcgcct aacctgtatg acggacctgc gcctcgtgcg ccgacatcac 1620 acagtcccag taaaagtcaa gactaccaag cagcccgatt ctattcttcg ccatcttcca 1680 tgaacagcgc cctacccgct tccccgccta cgacccgccc agctgttgcg ccttttaacc 1740 cttccgtcat ccttccatct ttcctctacc tcggccctga catccaatcc gaatccgatg 1800 ttcaatatct tttccgatta ggtgtgaagc ggatattgaa tgtcgcgttg gaatgtgatg 1860 acaatcaggg attgagcttg aaagagaggt tcaagtatag aaaagtgggt atgagagata 1920 tcgtggaaga aaacggggtt gggaagggca tgagagatgc ttgtgaattc ttggatgatg 1980 ctcgccttca ctctgcacct acctacgtcc attgccaagc tggcaaatca cgttccgtca 2040 caataatcct tgcttacctt atccatgcca acgcatggac tctcaaaaca tcctatgctt 2100 atgtcgcaga gcggcgaaag gggattagcc caaatatcgg tttcgtcgcc gagctgatgc 2160 agtgggaaga gaaggaattg ggagtcaagc agagtggggg cgtgcatgga gatgggaatg 2220 ggagggctaa agctccaggt ggtggaggcg gtggtggtgg ttcaaggcac atggaagatg 2280 gaggtgatga tgagggaaaa ggcaagactc atcttcggga tagtttaccg cctacctggt 2340 cgagtagtgt ggatacttat acccgtccag ccaaggtata ctccccagtg ggcagggacg 2400 atggtggaga agaagaaagt ggaagggaag gaaggattgc agtgggcgac gaaagggagg 2460 tgaggaaaaa tggtgtctgg atgcatcatc ggagagcacc tgtggatcga accacccttc 2520 aacccggccg acgagtctcc aaagccggtc tcgaatccct tcgaccattc ctgattacct 2580 ctaccgatgc ctcctcctct tctgccgcgc ctaataatgg tgacaatatc gatagtgaac 2640 gtcaagtcaa taacggctca gaggcgagac cgtcgcctag ggccagcccg gggatgggta 2700 tgggagggca tgcgatgacg cctgccgggg atggaccctt gaagtggata taaatctctg 2760 atgatgcgct aaatgggcga ggagggcaga gagagtgatg atctattggg acgttttaac 2820 gagccattgc gcttctacta tctactggtt gcataattta tgggttgttc atggactgta 2880 taaatataat ttataattta tcagttgtgt atatgtatat ttcttcttgc c 2931 <210> 36 <211> 1016 <212> DNA <213> artificial sequence <220> <223> OCA1 <400> 36 gttcattgag cagattggaa agatcacgtg actcaagcga ttgtgtccat cacagagaaa 60 acgaatgcga agacaacgaa ggaaagcaac gttctcgatc gcataaaccc agcatcaggc 120 atacgctcac ataagagaca gcatggcgaa gatagtgcca cccatgaact ttggcctcgt 180 agaagacgga ttctaccgtt ccgcccagcc ttccgagctg tgcttctctt ttctcgagaa 240 gctaaatttg aaaagcatta tatgggtggg agctgaagag ccttcagaca tcttcttgtc 300 attcatcgaa tctcaaggga tcaagttgta caacctcgcc cctcaaacga gtttgaaccc 360 acatttccca cctccataca cagattcggg cgtagtacct atatctggcc aataccacct 420 tccaccactt cctcctccgc ccgaaccact gatcattcag gctctaactc ttctattacg 480 cccatccact ttcccaacct tattatgttg taatatggga cgccatagga cagggactgt 540 ggtgggatgt tacagaaagc tgcaacggtg ggcgttgagt agtatattgg aagagtatag 600 acggtacgca gggatgaagg tcagggtcct gaacgagcaa ttcattgaac tgtttgatac 660 agacttagtt tcaataacag cggaacaggt gacaaaatag taattgtccg cgacaaaagg 720 ctagcaaaat tcctttcatg ggaccagtag aagaagtcaa actgccaccg tgcccaactg 780 tagcccatac aacgcctaga gtgtctaccc gatcagggat gctggactgc gtctgcacca 840 tgcagcggcg atcattgcgt tgtggttgat aaaatcatta tccataagaa atacatacag 900 cacgaaatcc aatatctgat gtaccatata tgcaactaga atgcgacaac cttggctttt 960 ggcttcgcga cgacctcggt cctcttcttc gccgtcagcg gcacactagg cgcttg 1016 <210> 37 <211> 2392 <212> DNA <213> artificial sequence <220> <223> GDA1 <400> 37 tttgagcctc ttgtgaccgt ctcggcattc acgctgcccc tcccacccct atcaaagatc 60 cgccactatc ccttactccc cattgttaac tttgattcct cttcactttt tttttcaatc 120 tctctctctt tttttccctt tcccaaatat ataacaggtg catgcctgca cacttattcg 180 ctcgttctac cgatactact ttcccttcct ccatagagac atccacatca ggctctctgt 240 ccttttccct tttgtcgtct ctgagacaca gaccgtcagg taggatcccg aaaaccccaa 300 tttcaccaaa atcccccact tccgcttcta cttccacgac cgccgccatg ttctccacgc 360 gcaagtactc tcctttaccc acgagtgcga atggtcccgc aaggaaacgc actggtgccg 420 gtttgacagc atggaagcga tgggctttac tcgccgccat ctctgtggcc gtttcttct 480 tggtgtttag ccgcgctagt ggcggatctg aacaacaaca gatctacaat gaagaaaaca 540 cgtatacacc ctcgctggac gaggatgttg tgggagacgg tgatccgatt gactatagct 600 ctcctccctt ccgtcctgaa gactctgatg tggcccagcc gttggaccat gaagatggag 660 atgacgatgg tgtgattcat acgcttccca ctggcgacgc ttccaaccct catgatccta 720 cttctaccga agcccaggat gcttctgaag ccgagcaaga ctttaccaac gagtctgagt 780 ctgaatctcc gtccgaggct gaatcttctt tccccggatc attcgagcaa gacccggacc 840 ccgcttcgac aacggcctgt accgagcccg tatcttctga caagcctgtc gtgcagtacg 900 cgcttaccat cgacgctggt tccacgggtt ccaggatcca cgtctacaaa ttcaacaact 960 gcggtccgtc cccccagctg gagtatgaga cgttcaaagc tgtcaagccg ggactttcag 1020 catacgctcg tgacccgact gcggccgctg cttctcttga ccctttgctt gaggaggcat 1080 acagggtcgt tcccgagagt ttgcgaaagt gtacgcctgt ggaggtgaag gctactgccg 1140 ggttgaggtt gctcggccag caggagagtg aggctatcct tgatgaagtg aggaacaggc 1200 tcgagaccaa ctgggacttt acggtcagtg gcgagagggc tgtcgagatt atggacggca 1260 aggatgaagg tgtctatgcg tggatcactg ccaactattt gctcaacaag attggtgaag 1320 gcgccgaatc tgacgacacg ctggcggtca tggacctcgg tggcgcttcc acgcaaatcg 1380 tctttgagcc gaaattcccg gcggagtctg accaggcgct ggtggagggc gagcacaagt 1440 acgagctcac ctttggcggc aaggacttta cgctttacca gcactcttac ctcggctatg 1500 gtctcatgcg cgccaggcga agcgtgcaca accttgtcgc attcacatgg agctttggcc 1560 agggtgaggt cgagtggggg aacttgagcg aggatgtaca ggtgccgaac ccttgtttgt 1620 caaagggtat gacccggaga gtcgcgcttg atccgcctgg aaggcagact gtcaatgtta 1680 ccatgcacgg tgggaatggt aactttgagg cttgtaacag ggtcgtcgag ttggtcatgg 1740 ccaaggacgc tatctgtgaa gtcaagcctt gctctttcaa cggtgtttac cagccctctc 1800 ttctcgatac gttccccccgt ggccaactgc tcgcgctttc ctactttacc gaccgcatca 1860 agcctcttct cccatcctcc tcttcctcca cgctctccat ctctgagctt acctctatgg 1920 ccaaggacgt ctgcgccggc ccggacgcgt gggctgaccg atggggcagc gacgcgacgg 1980 cgatggagga gcttgccggt aggcccgagt actgcctcga cttgacgttt atgaacgcgt 2040 tgctcgggct cggatacgag ctttctccgg agagggagtt gatggtggag aagaagttga 2100 ggggtgtgga gcttgggtgg gcgttgggtg ccggtttggc gttggtggag aaggcagaat 2160 tgacttgtac tgcgtagcgt agcgtagcgt aaaactaaaa aaaaaaggag tatggttata 2220 tagagggggt ttagagtgga tggaaggaaa aaaggtcaat gggcccacag tcttttgtaa 2280 aaatcacatt agttttagat ctatacaaag atcatatcaa tcattgcaat tctttggtac 2340 tgttttggat aggatataca tggatgcacc tgtgtaagta caagatggga aa 2392 <210> 38 <211> 2125 <212> DNA <213> artificial sequence <220> <223> FBP26 <400> 38 cgttcacatc catccaatcc gttcttctca atttactgaa aacaatgtcc ataccaccac 60 caccaccgtc caataagtcc cccgcatcag cagtttcccc ctccaagccc cgctccccaa 120 agctcaaacc cctcactccg acatccgaaa aaccttcgcg cactaataat gacgacgatc 180 aagtctacca gcccgtcgag ccacatgtcc tggccgaagc agtctcaaaa ctcgacatga 240 tccgatcagc acctgcaccc atgtctactg tgacttctcc cgcggctagt gcagctccca 300 gtggtcctag ctcgccaaga ctctctggtg cgggccaggg ggcgccatca acaggtccat 360 gggctatgga ccgcacagcg agtggagatg gtaggcacag tgcgcctggt acacctcact 420 tcggggcctc aaccgctttg ctgaagacgc tggatgagac cacgaagggg atcaggcaaa 480 gctccagggc tccgtcgcgt gcgccatccg tgtctggtat cggtactgtc gttgaaaagc 540 ctgactattc cgaagccaag atcgtcgtcg cgatggttgg tctcccggcc cgaggaaaat 600 cttatctcag taacagactt atgcgatacc ttcgctggct cgaatacaac gttcaagtat 660 tcaacgtcgg acaactccgt cgctccaaag cccgctccgc tctccaggcc gggcagggaa 720 aggtcgacca ttccgcgaca tacttctcgc attcagatgc ggaagctacc aagaaacgag 780 aagaactcgc agaagagtct ctcgaatcac ttatttcttg gctgaagaag gaaggaaatg 840 tgggaattat ggatgcgacc aatagtacaa tcgatcgaag ggagaagatc aagtcgcgaa 900 tcgacaagga gccggggctt caagtcctat atcttgaatc tttctgcgat gaccccgtag 960 taattgcaac caacattgca ctcaaggtcc gatctggcga tcctgactac caagggatgt 1020 ctaaagaaga cgcagagcgg gattttagga agagaatcgc tcagtatgag agtgtatatc 1080 agacgatcaa cgagccaaat attcccttct gcaggatatt aaatgtcgga cagagagtca 1140 cgataaataa gattgagggc tatcttcaaa gtcgaattgc attctatttg atgaacctgc 1200 atctcaaacc aagaagtatc tatttgtcaa gacatggaga aagtatgtat aatgtcgagg 1260 ggaagattgg aggtgattcg gatctctcac caagaggatg ggaggtatgcc cgcgcacttc 1320 ccgctcttat caaagacaac attggcgagg ggcctcttga ggtctggacc tcaacccttc 1380 aacgtaccca acaaacagca tcataccttc ctttcgagaa gaaaacgtgg aagtcgctcg 1440 acgaactgga cgccggtgta tgtgatggca tgacgtacaa ggagatcgag caaaagtatc 1500 cagaggatta tgagagtcga gacgacgaca agttcaatta cagatatcgt ggtggagagt 1560 cataccgtga tgtcgtggtc cgtcttgaac ccgtcatcat ggaacttgag aggcaaaaca 1620 atattttgat tattgcccat caggccatcc ttcgatgtct atatgcctat ttccaggcca 1680 gaccccagca agaactccca tacatcaaca tccccctgca cacacttatc aaaatcacgc 1740 ctcaagctta tggctgtcaa gaagaacgct accctctccc tatcgctgca gtagacaccc 1800 atcgaccccg tccatccaag gggagaaaca ctgccggtgt ttcagtggcc gaagaagcct 1860 tccagcctgt caagcgagat tattatggag acagccaaca aggtgtcggg tttggcttga 1920 agcccgaggc gatttcacag gctttggaga acgagatgga acaaggaaag ttgacaccaa 1980 gggctgcggt agctgcgcaa ttgcatcacg agtgatgaat cttattggga gttgtaagcg 2040 tagaattaga tgtatcgaga agcccagaaa tgaaatgcaa ggcatgatgt aatgaagtct 2100 tcctccgatg tatgctctgg tcgcg 2125 <210> 39 <211> 2418 <212> DNA <213> artificial sequence <220> <223> PSR1 <400> 39 ttgctacacc aggaaggaag gaagaacgcg caggaaagaa gaaagaacga acgagagaaa 60 gaaagaggca gtagaagaaa tgcccacaac gcgcacagag ccccccactg tggcaccagc 120 catcagtccc caaaacacaa acaccgccgg acccgcacac acaacatcca tcgaccacaa 180 cacgtcaacc accgacactc agcaaccgtc ctccggcttg caaccttcca tcctgcctcc 240 tgtagccact ccagcgacac agaatctagc ttctactaca gaaatgacca aggacggagg 300 tgcagctgca gcccagcctt ccacggctca gacaactctg cctgaacccg gtacaacgtc 360 tacatccatc aagcctacag aaggagagca aagcaaaggt acccccctgg gaaacttgtc 420 ccgtagatta tcaaacaagt ccccatcgac taccgcttcc tctgcacccc aaacaacggc 480 tgaaaaagca gacccaaaac cggcttcatc acacactcag cctaccacct ctacgtcaaa 540 aacaacggtt aacacgcctg catcccgcag tgtcaatgga gccacaaagt ctaagacagc 600 tcctacatcg aatacaactg cgcccaaggc cgggcaaaag aagaaaagga agcggaaggg 660 tttggcgggc attttactcg cgctcggatg tttatctgtt gatgagtttg aggagggagcc 720 aagcaaacct agcagcacga ctgcgagtgt aggggcgggt aaaactgccg gcgctggcgc 780 tacaacgggg gtgagcacaa aagccgatga gagcgccaaa ccagggtcag gtgatgctgg 840 catgacctca ggtgccttga aggcaccgaa cggtagcgtc gcacctgctc cgtcaggccc 900 atcagcagtc aaaactcaag acaccactgt aggagctgaa caaaaggtgg atgcaaccgg 960 cccaaccggt tctacggttg ttgctgaagg atcgaatgaa gccgataaag gtatcgtccc 1020 cgatgaacaa gtcgtcgtgc ctccgaccga acctcatacc cttccagatg atgagaccgc 1080 tggtgtaacg tcttctgcgg tccagcctcc tggaggaggc tctgtcctcc ttggcacccc 1140 gtctaaacac gtctctcacc gcgaatctga aaccaacctt ggtacacca gtaatgagcg 1200 tacagagaca agcgggggat actcggacat tagcaattct gaaatggttg acgaaagcac 1260 aggacaagga ggagatgaac tcggagaaga ttatcttgag tatgatgacg aagaagatcg 1320 attaattgaa caaggtgggaa ttggaattcc cgtggacgag aatggcaatc cggcaccatt 1380 attacccccc atagctgcca agcaccgtgg acgaaagtgt ctcgtgctcg atctcgatga 1440 aaccctgtta cacagtagct tcaagggcga aaatcagcaa ttgcccacag cggattacat 1500 tgtaccggta gagattgaat ctcaagtgca caacgtttat gtcatcaagc gaccgggtgt 1560 cgaccacttt ttgacagaaa tggcaaagat atatgagatt gtcgtgttca ctgctagttt 1620 gtccaagtac gctgatcccg tccttgacat gcttgacgag aaccgtgtcg tagcccatcg 1680 tctgttccgt gaaagctgct acaaccaaa aggaaactat gtcaaagatt tatcccagct 1740 cggtcgtgac atccaacact ccatcatcat tgacaattca cccgcctctt acatcttcca 1800 ccctaataac gccgtccctg tgtccacttg gttcagcgat ccccacgata gtgaattgac 1860 cgatctttgc cccttccttg cagacctcgc cactgtcgac gacgttcgtg gtgtccttga 1920 tggacgaatc tagacgttct agaaagcatc tcaacttata tccgcgagga ccatcgatac 1980 ccatcttccc ttcttttttg ggatcaggtc agggaaaacat tagagtgtat aaggactcat 2040 acggtaaact tggttgccca acctagaaaa tcataacatc aataacccta tacccagagg 2100 cctttcggga gcttggtcac atattcagtt attttctcta atctcatttg gttcatgtta 2160 cgatctcaag gcgtatgatc tattatgtcg atccctcatg gtctgttttc taatcttttt 2220 tattttcgtg ggtgtataca acctaggtac ccgcgcttat atctatcata ctccctcatg 2280 actgacttct tttttatctt tgctttagtt tcaaaatgtt gagtattct ggaatcgctt 2340 gtagatacgt acaaaaaat aattgggaca aagattgcat tgattgggct cgtcgcatgc 2400 atcgtcgtca tacagtac 2418 <210> 40 <211> 2684 <212> DNA <213> artificial sequence <220> <223> CDC1 <400> 40 gcgtcttgtt ccctggctga gggccggact tggaactagc ttctttttct tatccatttg 60 ctgcttgatc catctgtttc cactagtcac aatgctcgga tcaccgcggg cacactcacc 120 cgcccctccc ctggccgcca gaggaaggaa gacaggcctc aagtcacggg ccacccagat 180 acttgcactc agattcggct gggtcgtgct cgttatatgg tacgaggtcg gagaattctt 240 ccactctctc tccacatgcc gcttccccga ctctgctctg cggcaggccc atccccaagc 300 cccgcctccc acccatgtcg tcctcatcgc cgatccgcat gttccccatg cacgactgtc 360 gtacccctcc gggaacccct ggctgaactg ggccaagcag cagatggatg agctcttcat 420 gcgcaaaagc tggaatgtgg ttatgcgtct agggagggta gaccaggtgc ttgttctcgg 480 agatatgttg gactcgggaa ggggggtcat gtccgatgaa gagtacgtgg agtacatcgc 540 tctattccga tcaatattcc agcttcctcc cacaacgcct atgcactttg taccaggtaa 600 ccacgacatt tctctcgtcc ccaatggcag attctcctct caagctcggt tacgctacca 660 gcaacatttc aaaacgccca acaccgtcct tcccatatca aaccactcgt tcattttgct 720 tgatgccgta ggtttggtgg aagaagacta tcggcggtat gcttcagaga tgcagtttgg 780 ggaatgggat ggtgtcaaag gcggtgtgat tgagtttgtc aaagacctga gggacaatcc 840 tccacctgga cccaaaatct tgctttccca tattccactg gcgagacctg aaggcgccgc 900 ttgtggaccg ctgagagaaa aaggacggat atcaaaaggt gctgggcctg gataccaaaa 960 tttgcttggg agtgagactt ccaaattctt gttggatgcc attcagccga acattgtgtt 1020 tagtggggac gaccacgatt actgtgatta tgtccacaaa gggaatatcc gagaagtgac 1080 agttaaatcc ttctcctcgt ccacgggcat ccgccgtccg ggactccaac tgctctcgct 1140 agttcctcca ccgacagaat ccaccgctag acttctccca acccacgccg accgaccttg 1200 cttccttccc gatcagctgg gtgtctattg gcgtgtctat ctccctctcg ccatccttac 1260 agcactgtac ctttttatca caaaccttcg ttcagcgtat ttgcgatggg accgttcttc 1320 acacgccgtt tcggagaaaa tgcggtcaag ccccgctttg ctctccgcgg agaccatgtc 1380 gcccaactca ttctcgtcac ggcggaatgg acctgtccca cttagtattc cctctcgtaa 1440 atcatcttca catctccccc tttctgctcc ctcagccatt ccctcttcca ccctgcctcg 1500 ccctgtacga tacaactcca ctcctgcaga gtacccgcca ggttccagga gcggccagag 1560 caaccctgta tccccatttg gaagtccgaa attgtccgcc gttgagcgtt ttggggagcg 1620 cgatgtagaa cgcgatggag aagcagcgtc tgctagtgtg acaggtctta acacacctct 1680 cacctgtct cggcgatcat cctacatata catggatcgc ggctttccat cttcagtctc 1740 tgactctgcg ccgttatctg cttcgggcac cactaactgg gggttgggtg caaatacggg 1800 agtcagttca ccctcatcgt cgggttttat tcggagagtt tctagcgcca acctgtcgac 1860 gttgatcact acgaatgtcg cccctccgag cttgagcatc acctcccccg gtacaccccg 1920 gcgtgtcacc ctccccagcc ctctccttct cccgcattcc cctgctcacg cgcaagccca 1980 ccccctctcc caaacctctt cacacgccac gcacccccat ccagctgtga tttacacttt 2040 ccccacccca tcaaggtcat ggttctggtt tgagagggcc aagtcgtttt taagatgggc 2100 atggaaagca aggaagggag ccgtgggcaa gagttggaga gagctgatca gtgttgcatg 2160 ggttggggct atcgtctggc taggtgtgaa tgcgttgttt ttccttgagt aaaacactct 2220 cgagtagaac gccgtggtgc ttgggtgctt ggggtgcttag atgcttgggg tcttgtgttt 2280 attgtttttt tttcttcgag ggcgtgtgga aagatgtcgc ctgcgttgca gcagggtgta 2340 tccaaaaaaa aattgtctac tgtctcttcc tgttttttcc ctcctagatt tgatctcttc 2400 ggtgtccgga agacacattt tctcactttg catcataaaa agagaccccg ttcttctaaa 2460 ggagatgagg gtcggttggc cgtggcctca tagcctcata acatatggat tttggagcgt 2520 gtagcgtaca tacatactgc tgttttacat gacctgcttt ttccgcacat ggtatctggt 2580 attcatgtac ggcaatatac tagttctccc aatcgccctt cacgaacgac cacgtgaagc 2640 tcttcaaatg ggactgactt tgcgccacgc agtgacgaag cccc 2684 <210> 41 <211> 1635 <212> DNA <213> artificial sequence <220> <223> INP5202 <400> 41 ataaaaatgg tgactctcta cttgccagcc aagcgttcat atctatgacg cacgcagttg 60 ttattgtacg taccccgccg cctcgctacc agacgagacg gatgttattt taactcttta 120 caacataatt gactttatca ttactttcag acaatcctca tccaagccac ccctaccgtc 180 ccaagttcaa ccccaagatg gctcctctag acgtcttcat gactacctgg aacaccggtt 240 tacagggatc caaagcccaa tcccaagacc tcacgagctg gcttctgcca gttttgcgca 300 atgcttctaa tcctgaactg cctcaagggt ttattccaga cctttacgct ataggaattc 360 aagagctttt acccctccat cttgctatgg ctggtttgac tgagcctgta ctccttgctc 420 tgactagtcg cattgaaaat ctcttatccg cccacgccag ttctatatcg cccaataaga 480 caccggagag atattcctta gttgccagag tggcccatgt tggaaatgca ctttggatct 540 tttcccggga tagtaccatg gatggaaggc taggaaaacc gtcgaccgca actctgggat 600 tatattgggg aggcatggga aataaagggg ccgtgggtgt cagactccct gtgagaagag 660 gcaagattgg tggatggggag aatctgacct tcgtcaatgc ccatctcgaa gcgcatgacc 720 acaatattcc tcgacgcaat gcacagtatc agaggattct gagctctctc gtcttcaatt 780 cgacggatcc tcttacaact tcacagcaga tttttgacac ttctcattta ttctttatgg 840 gtgatctcaa ctataggctt tccaagcagc cacctccagg agctctacaa gagaacaaaa 900 tgttcggaga tgtgctggaa ttggaaaagt ctcgaatggg gatgctggat actgatactt 960 tgagacaaga acagagagaa gggagggttt tcggaggctt aagggaaggc gatttgacta 1020 gattcgcgcc cacgtacaaa aggatagttg gacaaattga aggatacagc aagaaacgca 1080 tccctggctg gactgaccgt attctctttg cttctcatac cgatcctccg catctctttt 1140 ctcccgaggc gtctttggac ccggtcccct ctaacgtagc cgatacaacc agtatcctcc 1200 atttcaactc cacaatcgag cttgtcattt ctgatcacaa gccagtccat gcgatccttt 1260 ctttaccgga agtttcccac gaagcgccct ctcctcacct tgcaccaacg cttccccctg 1320 ctccttcacc tcatcagcca agacctcttc ccactcagcg cgaggtcctt ttgattgaga 1380 agtttttggg gactttgctc gatagacttg tcgggtggcc atggtgcatt atcgttctgc 1440 ttgggtttgg caacacacgt aggggcatgg gtgtgagcgc ctttgtggcg atgatttggg 1500 gcatctggtg gagcggggta tactctggat gattctgtaa attgataaag gcttctgtct 1560 ataggatact tgatgtatgg atttttcgtc gttcttatac atatataatg tactcatcgt 1620 aataacgaag tgagg 1635 <210> 42 <211> 3400 <212> DNA <213> artificial sequence <220> <223> YND1 <400> 42 gttactttgc tataccattt aatactccac ctcccccagt ttccgaacaa caagtctgga 60 cgagggtgga cgacacggga agatacgagg aaagaagagg tccttcattc caaggcagaa 120 aacgagaccc gcatatcgtc ataactgccc ccaccccacg cacatctccg ctccgcagca 180 aaagaaaacg tacaacatct tggctaacaa aaaagtcgca tacggggcga ctccagaacg 240 ataagatact gcgcacttag ctcccataca catctcattc agatttgtcg agtgcataca 300 taggagccag ggcacaagca cgatggcacc atccgttaca ccactcacca ctcactatgc 360 cctagtcatt gacgccggct catcgggttc gcgattacag atatattcct ggcgagatcc 420 agatttggaa agagcagaaa ttctccagga tgtgcagaac attgagagac aaggttcgag 480 ttcgagctcc aaggaaggcg ctcggtggtg gtggagtgga gaagatggat ggaagggcaa 540 agggaaaggg aaagcgaagg agatggagga gatggctttg agaaggttgg tgagagttgg 600 caaaggggtg gaaggggatg actgggtgaa aagagttgag cctggtatat ccactgtcga 660 ccccgagaat atcccagagt acctcgcccc gctgctcaca catgctctcc aacacatccc 720 accctcggtc cactcctcca cacccatcta cctcctcgcc acagcgggca tgcgtctctt 780 gccttctcag cagcgcgatg ccattttaca agctacgtgc gatacattac gaaacgacta 840 cccgtttctc gtctctgggc cgacagagga agggccttgt ggtgaaaatg taagggtgat 900 tgatggagag gaggaaggta tctggggttg ggtcgctgtg aactatctca tggacggttt 960 cggtcatgcc ccttcgcctt catctatatc caattctgga acatcatcat cgtctagtac 1020 caacctgctc cctctcgccc cgttagcttc tgcccctcca gactcttcct cctcttccat 1080 cacccccgtc gacattgccc accactcacc cacattcggt ttcctcgaca tgggcggcgc 1140 ttccacccaa ctagctttct ccccctctgc ttccgaactt ctcacctctg gtttcccgct 1200 cgacaaactc cggacagtta gtctcagatt actttcgggc gaacaagtcg attggccggt 1260 ctttgtagcg agctggcttg ggttcgggac gaaccgcgct cgggaacggt atatgacttc 1320 tctctatcaa caatgggcct ctgcccatcc ttccccttct gcacaagacc tagcgacacc 1380 cattccggac ccttgtctcc ccaaagacct ctccatcctg ccaccctcct cttctcaacc 1440 ccctttaatc ggaaccggct cattccccga atgcctcacc tccctccacc ctctcctcga 1500 acattccacc ccttgcccca cctcccactg cctcttcgga ggccaaccga cgcctcacat 1560 tgattttgaa cggcacgatc agagagggtt tatagggata agcgagtatt ggtatacgat 1620 gcagcatgtg ttgggggtag gaggggtatg ggattggggg gaatgggaaa agggaatgaa 1680 gggagttttgt gggaaggatt gggaagtgat taaaagtgaa gtggagaatg gggattggga 1740 ggatgttaat atggacccga caagattaga aatgcaatgt ttcaaaggcg cctggatctc 1800 caacgtctta cacgaaggga tcggtatccc ccgactagtc gatgtaggcg ggaatgatac 1860 actcacaggc gggtcgttgg gcgatacgaa cgctgaagct gaacgccggg ctcgtgaaaa 1920 gggtctgttc gaaaagaaag ggcaaggaca agggaaacat catttccaaa gtatggacca 1980 agttgggggaa acagccatct cttggacact cgggaaggtc gtgattgaag catcgaaagc 2040 cgtccaacct cgatcgcaag aaatggaagg gtggtggatg cgtcatctca acctcgggtc 2100 catgcgactg ccgttatcgc taccgatacc aaaacaccta gagggaaagc ttgaagatct 2160 aggattgagc gtggtatgga tttatgcggt agtggggttc tttttggtgg ggatgctctt 2220 ctctcgttct aatcgtcgca gggggagttgg ttcgttgggg tctgggatgg ggagaaggag 2280 aaaaccaagt ttgtcctcac ctcctttacc tgcccgaccg tggttcactt tcccttcttt 2340 cttctccggc cccgccgccg atccgagctt atcgatcgaa gacggcccag acgcttcccc 2400 cacatcatcc acgtcatcca cccctttctc tggcaatggc accgctggtg gtgctagtgg 2460 taaatcgcgt atcgtccccg gccgactccg actttggtcc ctccgtatct ccaacacaat 2520 caacaaatat attcctgcct cacttccttt atctcttggc agcccgaact cgagacagcg 2580 gggaggcgca catgagctgt ggacatcgat agggataggg ttaccgagaa cgcgacataa 2640 ttcgatgccc atgatcggca tgggaccgaa tacttcccca cgtgttggtc ttctctcccc 2700 gggtggtgac ggtggttact cacagcccgg ttcacctcgt atcatctccg caccattctt 2760 catccccgcc gctgctccag gaatcggtgg cctcaacacg ggtgtgggta gtcttacacc 2820 agaaacagtt ctgacaggca tatcaagcgc cacatctgtc tcaccttctc cgagcctcgc 2880 atctacttct tcgccgcctc cgcctaggag cagtttgaaa cctggcaagt ctggtcgacc 2940 gttcaaaccg aggcagaatt caaataatct gcatccacac catggatcgc atgggtttca 3000 ttccgttggg gaaggtatag gtgcaggagg gggagggtgg aatgatccgc ctttggccat 3060 gttgagtagc cccggttcgg gcacaggtcc aagcgggagc ggagcggcag atgatggtgg 3120 cgtgttgacg ccgacggcga atggaggctt gagtaatggt gcattgtcga ggaattctag 3180 tagggcgaat ctgagtgaat tggggttggc acagcggtcg atgagtagaa ccggggacacc 3240 tggttttgat taacctcttt agatgcgcga tataataagg acattcaggg ttgttttgtt 3300 gttggaaaat attcatctag ccccacaagt atatacatat gcacgcctgt tatattattc 3360 cttatcgaag gcactcctta catgtacgtt atatacagga 3400 <210> 43 <211> 20 <212> DNA <213> artificial sequence <220> <223> VPS29 L1 primer <400> 43 tgtcttcttg aaaggggttg 20 <210> 44 <211> 38 <212> DNA <213> artificial sequence <220> <223> VPS29 L2 primer <400> 44 tcactggccg tcgttttaca atgacgagaa ccaggacc 38 <210> 45 <211> 40 <212> DNA <213> artificial sequence <220> <223> VPS29 R1 primer <400> 45 catggtcata gctgtttcct gactcagtct caatcaacgc 40 <210> 46 <211> 20 <212> DNA <213> artificial sequence <220> <223> VPS29 R2 primer <400> 46 gttgtttctt ctttcccagc 20 <210> 47 <211> 20 <212> DNA <213> artificial sequence <220> <223> VPS29 SO primer <400> 47 aatcaggcgt gggttcagac 20 <210> 48 <211> 19 <212> DNA <213> artificial sequence <220> <223> VPS29 PO primer <400> 48 ataggtctcc ttgtcgcag 19 <210> 49 <211> 21 <212> DNA <213> artificial sequence <220> <223> VPS29 STM primer <400> 49 acacctacat caaaccctcc c 21 <210> 50 <211> 24 <212> DNA <213> artificial sequence <220> <223> VPS29 STM common primer <400> 50 gcatgccctg cccctaagaa ttcg 24 <210> 51 <211> 19 <212> DNA <213> artificial sequence <220> <223> YMR1 L1 Primer <400> 51 tactatgtgg gcgagaagg 19 <210> 52 <211> 38 <212> DNA <213> artificial sequence <220> <223> YMR1 L2 Primer <400> 52 tcactggccg tcgttttact gctactcgta atgcgtcc 38 <210> 53 <211> 39 <212> DNA <213> artificial sequence <220> <223> YMR1 R1 Primer <400> 53 catggtcata gctgtttcct gagcaacaaa aggcttggg 39 <210> 54 <211> 21 <212> DNA <213> artificial sequence <220> <223> YMR1 R2 Primer <400> 54 gcttgttctc ctgaagtttg g 21 <210> 55 <211> 21 <212> DNA <213> artificial sequence <220> <223> YMR1 SO Primer <400> 55 ccagacaaaa ggctcttagt g 21 <210> 56 <211> 19 <212> DNA <213> artificial sequence <220> <223> YMR1 PO1 Primer <400> 56 agacttccac ttttaccgc 19 <210> 57 <211> 21 <212> DNA <213> artificial sequence <220> <223> YMR1 PO2 Primer <400> 57 gtcggagaaa aagagtagtc c 21 <210> 58 <211> 20 <212> DNA <213> artificial sequence <220> <223> YMR1 STM Primer <400> 58 tgctagaggg cgggagagtt 20 <210> 59 <211> 24 <212> DNA <213> artificial sequence <220> <223> YMR1 STM common Primer <400> 59 gcatgccctg cccctaagaa ttcg 24 <210> 60 <211> 22 <212> DNA <213> artificial sequence <220> <223> SSU72 L1 Primer <400> 60 gactatgatg aaaagacggt cc 22 <210> 61 <211> 39 <212> DNA <213> artificial sequence <220> <223> SSU72 L2 Primer <400> 61 tcactggccg tcgttttact gggaggcaat gaggatgac 39 <210> 62 <211> 41 <212> DNA <213> artificial sequence <220> <223> SSU72 R1 Primer <400> 62 catggtcata gctgtttcct gtatcctcaa cgctcacggt g 41 <210> 63 <211> 18 <212> DNA <213> artificial sequence <220> <223> SSU72 R2 Primer <400> 63 aaccttggtc tccttgcg 18 <210> 64 <211> 19 <212> DNA <213> artificial sequence <220> <223> SSU72 SO Primer <400> 64 atctccgttc aggactgtc 19 <210> 65 <211> 19 <212> DNA <213> artificial sequence <220> <223> SSU72 PO Primer <400> 65 actgggagga tagtttggc 19 <210> 66 <211> 24 <212> DNA <213> artificial sequence <220> <223> SSU72 STM Primer <400> 66 ctccccacat aaagagagct aaac 24 <210> 67 <211> 24 <212> DNA <213> artificial sequence <220> <223> SSU72 STM common primer <400> 67 gcatgccctg cccctaagaa ttcg 24 <210> 68 <211> 19 <212> DNA <213> artificial sequence <220> <223> NEM1 L1 Primer <400> 68 atctatgcca ctgaaagcg 19 <210> 69 <211> 38 <212> DNA <213> artificial sequence <220> <223> NEM1 L2 Primer <400> 69 tcactggccg tcgttttact atgcgactca gggtgttc 38 <210> 70 <211> 40 <212> DNA <213> artificial sequence <220> <223> NEM1 R1 Primer <400> 70 catggtcata gctgtttcct gtgtgaatga tgtgcggagg 40 <210> 71 <211> 20 <212> DNA <213> artificial sequence <220> <223> NEM1 R2 Primer <400> 71 agaggaggat ttggcttttc 20 <210> 72 <211> 18 <212> DNA <213> artificial sequence <220> <223> NEM1 SO Primer <400> 72 tcacgagcct ttttgtcc 18 <210> 73 <211> 19 <212> DNA <213> artificial sequence <220> <223> NEM1 PO Primer <400> 73 acgctgatgg aggagattg 19 <210> 74 <211> 20 <212> DNA <213> artificial sequence <220> <223> NEM1 STM Primer <400> 74 caccaactcc ccatctccat 20 <210> 75 <211> 24 <212> DNA <213> artificial sequence <220> <223> NEM1 STM common primer <400> 75 gcatgccctg cccctaagaa ttcg 24 <210> 76 <211> 18 <212> DNA <213> artificial sequence <220> <223> YVH1 L1 Primer <400> 76 tgctcatcca ttctcagg 18 <210> 77 <211> 38 <212> DNA <213> artificial sequence <220> <223> YVH1 L2 Primer <400> 77 tcactggccg tcgttttact atggctgtgc gacttgag 38 <210> 78 <211> 40 <212> DNA <213> artificial sequence <220> <223> YVH1 R1 Primer <400> 78 catggtcata gctgtttcct gcacaaaaac tgacgctgag 40 <210> 79 <211> 19 <212> DNA <213> artificial sequence <220> <223> YVH1 R2 Primer <400> 79 ctccagattt tgtggcaag 19 <210> 80 <211> 19 <212> DNA <213> artificial sequence <220> <223> YVH1 SO Primer <400> 80 tctttgccga cttccactc 19 <210> 81 <211> 18 <212> DNA <213> artificial sequence <220> <223> YVH1 PO Primer <400> 81 gaccttttca gcgattcg 18 <210> 82 <211> 20 <212> DNA <213> artificial sequence <220> <223> YVH1 STM Primer <400> 82 tgctagaggg cgggagagtt 20 <210> 83 <211> 24 <212> DNA <213> artificial sequence <220> <223> YVH1 STM common primer <400> 83 gcatgccctg cccctaagaa ttcg 24 <210> 84 <211> 22 <212> DNA <213> artificial sequence <220> <223> OCA101 L1 Primer <400> 84 ttctgatgct cacactactc tg 22 <210> 85 <211> 39 <212> DNA <213> artificial sequence <220> <223> OCA101 L2 Primer <400> 85 tcactggccg tcgttttaca ggaggacagc atacaaatg 39 <210> 86 <211> 40 <212> DNA <213> artificial sequence <220> <223> OCA101 R1 Primer <400> 86 catggtcata gctgtttcct gcgaaacttc cagtagcctg 40 <210> 87 <211> 18 <212> DNA <213> artificial sequence <220> <223> OCA101 R2 Primer <400> 87 gaacgacgga ataatggc 18 <210> 88 <211> 20 <212> DNA <213> artificial sequence <220> <223> OCA101 SO Primer <400> 88 gacgaggtgg tggaagatac 20 <210> 89 <211> 18 <212> DNA <213> artificial sequence <220> <223> OCA101 PO Primer <400> 89 cttgagttct gccattcg 18 <210> 90 <211> 20 <212> DNA <213> artificial sequence <220> <223> OCA101 STM Primer <400> 90 ctagagcccg ccacaacgct 20 <210> 91 <211> 24 <212> DNA <213> artificial sequence <220> <223> OCA101 STM common primer <400> 91 gcatgccctg cccctaagaa ttcg 24 <210> 92 <211> 19 <212> DNA <213> artificial sequence <220> <223> SIT4 L1 Primer <400> 92 agtgaggtag aaaccacgg 19 <210> 93 <211> 37 <212> DNA <213> artificial sequence <220> <223> SIT4 L2 Primer <400> 93 tcactggccg tcgttttact gaagagattg ggatggg 37 <210> 94 <211> 40 <212> DNA <213> artificial sequence <220> <223> SIT4 R1 Primer <400> 94 catggtcata gctgtttcct gactaatctg tcctggctgg 40 <210> 95 <211> 20 <212> DNA <213> artificial sequence <220> <223> SIT4 R2 Primer <400> 95 caagggtcta aaggaagtcc 20 <210> 96 <211> 20 <212> DNA <213> artificial sequence <220> <223> SIT4 SO Primer <400> 96 gataccccaa gtgtccctac 20 <210> 97 <211> 20 <212> DNA <213> artificial sequence <220> <223> SIT4 PO Primer <400> 97 cattaccgca tctgtagcag 20 <210> 98 <211> 18 <212> DNA <213> artificial sequence <220> <223> SIT4 STM Primer <400> 98 ctttaaaggt ggtttgtg 18 <210> 99 <211> 24 <212> DNA <213> artificial sequence <220> <223> SIT4 STM common primer <400> 99 gcatgccctg cccctaagaa ttcg 24 <210> 100 <211> 19 <212> DNA <213> artificial sequence <220> <223> GUA1 L1 Primer <400> 100 tcagtcttgc tctcttcgg 19 <210> 101 <211> 38 <212> DNA <213> artificial sequence <220> <223> GUA1 L2 Primer <400> 101 tcactggccg tcgttttacc aagatgagga tggtgtcg 38 <210> 102 <211> 45 <212> DNA <213> artificial sequence <220> <223> GUA1 R1 Primer <400> 102 catggtcata gctgtttcct gattacttcc aagcctcctg ggacg 45 <210> 103 <211> 18 <212> DNA <213> artificial sequence <220> <223> GUA1 R2 Primer <400> 103 tgcttgcgta tgacagac 18 <210> 104 <211> 19 <212> DNA <213> artificial sequence <220> <223> GUA1 SO Primer <400> 104 gggcatctgt atgtttgcg 19 <210> 105 <211> 19 <212> DNA <213> artificial sequence <220> <223> GUA1 PO Primer <400> 105 cttaggcttc caggacaac 19 <210> 106 <211> 24 <212> DNA <213> artificial sequence <220> <223> GUA1 STM Primer <400> 106 ctggggattt tgatgtgtct atgt 24 <210> 107 <211> 24 <212> DNA <213> artificial sequence <220> <223> GUA1 STM common Primer <400> 107 gcatgccctg cccctaagaa ttcg 24 <210> 108 <211> 19 <212> DNA <213> artificial sequence <220> <223> INP5201 L1 Primer <400> 108 cgtttccatt tggggtcag 19 <210> 109 <211> 38 <212> DNA <213> artificial sequence <220> <223> INP5201 L2 Primer <400> 109 tcactggccg tcgttttact gcgaggctct caaacttg 38 <210> 110 <211> 41 <212> DNA <213> artificial sequence <220> <223> INP5201 R1 Primer <400> 110 catggtcata gctgtttcct gcgaagcgag aaaaggagtt g 41 <210> 111 <211> 22 <212> DNA <213> artificial sequence <220> <223> INP5201 R2 Primer <400> 111 ttcttcctca cctggatacc gc 22 <210> 112 <211> 20 <212> DNA <213> artificial sequence <220> <223> INP5201 SO Primer <400> 112 agtcttgatg gcttcttcac 20 <210> 113 <211> 21 <212> DNA <213> artificial sequence <220> <223> INP5201 PO Primer <400> 113 gctgtttaga gtgagtagag g 21 <210> 114 <211> 24 <212> DNA <213> artificial sequence <220> <223> INP5201 STM Primer <400> 114 acagctccaa acctcgctaa acag 24 <210> 115 <211> 24 <212> DNA <213> artificial sequence <220> <223> INP5201 STM common primer <400> 115 gcatgccctg cccctaagaa ttcg 24 <210> 116 <211> 18 <212> DNA <213> artificial sequence <220> <223> PHS1 L1 Primer <400> 116 gtgggatggg aaatgatg 18 <210> 117 <211> 34 <212> DNA <213> artificial sequence <220> <223> PHS1 L2 Primer <400> 117 ctggccgtcg ttttacagtg acgcttttgt gtcg 34 <210> 118 <211> 36 <212> DNA <213> artificial sequence <220> <223> PHS1 R1 Primer <400> 118 gtcatagctg tttcctgtaa aggcaagacc gtaggc 36 <210> 119 <211> 19 <212> DNA <213> artificial sequence <220> <223> PHS1 R2 Primer <400> 119 ccttcgtctt cttcgtctc 19 <210> 120 <211> 20 <212> DNA <213> artificial sequence <220> <223> PHS1 SO Primer <400> 120 acactgaaga gactcccgag 20 <210> 121 <211> 19 <212> DNA <213> artificial sequence <220> <223> PHS1 PO Primer <400> 121 tgagtagcgg atgacttcg 19 <210> 122 <211> 20 <212> DNA <213> artificial sequence <220> <223> PHS1 STM Primer <400> 122 ccatagaact agctaaagca 20 <210> 123 <211> 24 <212> DNA <213> artificial sequence <220> <223> PHS1 STM common Primer <400> 123 gcatgccctg cccctaagaa ttcg 24 <210> 124 <211> 22 <212> DNA <213> artificial sequence <220> <223> MRE11 L1 Primer <400> 124 ggcatacttg ataaggaact cg 22 <210> 125 <211> 38 <212> DNA <213> artificial sequence <220> <223> MRE11 L2 Primer <400> 125 tcactggccg tcgttttaact accttggttg tgagtcgg 38 <210> 126 <211> 41 <212> DNA <213> artificial sequence <220> <223> MRE11 R1 Primer <400> 126 catggtcata gctgtttcct ggaatgaaga gggaatctgc g 41 <210> 127 <211> 20 <212> DNA <213> artificial sequence <220> <223> MRE11 R2 Primer <400> 127 tctcaaggtc gttgccatcg 20 <210> 128 <211> 19 <212> DNA <213> artificial sequence <220> <223> MRE11 SO Primer <400> 128 tggcttacaa gaactcagc 19 <210> 129 <211> 19 <212> DNA <213> artificial sequence <220> <223> MRE11 PO Primer <400> 129 cggctcatct cctatttcg 19 <210> 130 <211> 24 <212> DNA <213> artificial sequence <220> <223> MRE11 STM Primer <400> 130 gtagcgatag gggtgtcgct ttag 24 <210> 131 <211> 24 <212> DNA <213> artificial sequence <220> <223> MRE11 STM common primer <400> 131 gcatgccctg cccctaagaa ttcg 24 <210> 132 <211> 18 <212> DNA <213> artificial sequence <220> <223> DBR1 L1 Primer <400> 132 ttccaatcca acagtcgc 18 <210> 133 <211> 38 <212> DNA <213> artificial sequence <220> <223> DBR1 L2 Primer <400> 133 tcactggccg tcgttttaact ccgaaatgcc tgttgagg 38 <210> 134 <211> 40 <212> DNA <213> artificial sequence <220> <223> DBR1 R1 Primer <400> 134 catggtcata gctgtttcct gttactccca ctcgctaagc 40 <210> 135 <211> 19 <212> DNA <213> artificial sequence <220> <223> DBR1 R2 Primer <400> 135 ttttggtagg tgggagagg 19 <210> 136 <211> 19 <212> DNA <213> artificial sequence <220> <223> DBR1 SO Primer <400> 136 agaaatagaa aggctggcg 19 <210> 137 <211> 19 <212> DNA <213> artificial sequence <220> <223> DBR1 PO Primer <400> 137 tactgaccct catactgcg 19 <210> 138 <211> 20 <212> DNA <213> artificial sequence <220> <223> DBR1 STM Primer <400> 138 tctctatagc aaaaccaatc 20 <210> 139 <211> 24 <212> DNA <213> artificial sequence <220> <223> DBR1 STM common primer <400> 139 gcatgccctg cccctaagaa ttcg 24 <210> 140 <211> 20 <212> DNA <213> artificial sequence <220> <223> SIW14 L1 Primer <400> 140 ctcattcagg atttaccacg 20 <210> 141 <211> 37 <212> DNA <213> artificial sequence <220> <223> SIW14 L2 Primer <400> 141 tcactggccg tcgttttaca aagttgttgg cgaggtc 37 <210> 142 <211> 40 <212> DNA <213> artificial sequence <220> <223> SIW14 R1 Primer <400> 142 catggtcata gctgtttcct gtcatcggga gtttgttcag 40 <210> 143 <211> 22 <212> DNA <213> artificial sequence <220> <223> SIW14 R2 Primer <400> 143 caactaccac tcacaactct tg 22 <210> 144 <211> 19 <212> DNA <213> artificial sequence <220> <223> SIW14 SO Primer <400> 144 gtttgtccag tttggcaag 19 <210> 145 <211> 18 <212> DNA <213> artificial sequence <220> <223> SIW14 PO Primer <400> 145 agtatgggat gattccgc 18 <210> 146 <211> 20 <212> DNA <213> artificial sequence <220> <223> SIW14 STM Primer <400> 146 ccatagaact agctaaagca 20 <210> 147 <211> 24 <212> DNA <213> artificial sequence <220> <223> SIW14 STM common primer <400> 147 gcatgccctg cccctaagaa ttcg 24 <210> 148 <211> 20 <212> DNA <213> artificial sequence <220> <223> SDP102 L1 Primer <400> 148 tctggtatct tcctcccttc 20 <210> 149 <211> 41 <212> DNA <213> artificial sequence <220> <223> SDP102 L2 Primer <400> 149 tcactggccg tcgttttact tgttgtgggt gtgttgctgg g 41 <210> 150 <211> 44 <212> DNA <213> artificial sequence <220> <223> SDP102 R1 Primer <400> 150 catggtcata gctgtttcct gcgcttctac tatctactgg ttgc 44 <210> 151 <211> 19 <212> DNA <213> artificial sequence <220> <223> SDP102 R2 Primer <400> 151 gttcctgttt ggatgcttc 19 <210> 152 <211> 19 <212> DNA <213> artificial sequence <220> <223> SDP102 SO Primer <400> 152 gaacacatcaag gctctccag 19 <210> 153 <211> 18 <212> DNA <213> artificial sequence <220> <223> SDP102 PO Primer <400> 153 tgtccaagga atggtctg 18 <210> 154 <211> 24 <212> DNA <213> artificial sequence <220> <223> SDP102 STM Primer <400> 154 ctccccacat aaagagagct aaac 24 <210> 155 <211> 24 <212> DNA <213> artificial sequence <220> <223> SDP102 STM common primer <400> 155 gcatgccctg cccctaagaa ttcg 24 <210> 156 <211> 18 <212> DNA <213> artificial sequence <220> <223> OCA1 L1 Primer <400> 156 aataaagaga gacgccgc 18 <210> 157 <211> 39 <212> DNA <213> artificial sequence <220> <223> OCA1 L2 Primer <400> 157 tcactggccg tcgttttacc gagaaaagag aagcacagc 39 <210> 158 <211> 43 <212> DNA <213> artificial sequence <220> <223> OCA1 R1 Primer <400> 158 catggtcata gctgtttcct ggtagaagaa gtcaaactgc cac 43 <210> 159 <211> 19 <212> DNA <213> artificial sequence <220> <223> OCA1 R2 Primer <400> 159 gcagaggaca gaaagcaac 19 <210> 160 <211> 20 <212> DNA <213> artificial sequence <220> <223> OCA1 SO Primer <400> 160 atcagatgaa cctgctcaac 20 <210> 161 <211> 19 <212> DNA <213> artificial sequence <220> <223> OCA1 PO Primer <400> 161 taacatccca ccacagtcc 19 <210> 162 <211> 20 <212> DNA <213> artificial sequence <220> <223> OCA1 STM Primer <400> 162 atgtaggtag ggtgataggt 20 <210> 163 <211> 24 <212> DNA <213> artificial sequence <220> <223> OCA1 STM common Primer <400> 163 gcatgccctg cccctaagaa ttcg 24 <210> 164 <211> 20 <212> DNA <213> artificial sequence <220> <223> GDA1 L1 Primer <400> 164 gctgtaggtg gcaaaggtag 20 <210> 165 <211> 37 <212> DNA <213> artificial sequence <220> <223> GDA1 L2 Primer <400> 165 tcactggccg tcgttttaca cgagcgaata agtgtgc 37 <210> 166 <211> 40 <212> DNA <213> artificial sequence <220> <223> GDA1 R1 Primer <400> 166 catggtcata gctgtttcct gtttggcgtt ggtggagaag 40 <210> 167 <211> 19 <212> DNA <213> artificial sequence <220> <223> GDA1 R2 Primer <400> 167 gcctttgtct ttctccagg 19 <210> 168 <211> 19 <212> DNA <213> artificial sequence <220> <223> GDA1 SO Primer <400> 168 agaaacgggg gaaacgaag 19 <210> 169 <211> 19 <212> DNA <213> artificial sequence <220> <223> GDA1 PO Primer <400> 169 ggaagtagaa gcggaagtg 19 <210> 170 <211> 20 <212> DNA <213> artificial sequence <220> <223> GDA1 STM Primer <400> 170 atagctacca cacgatagct 20 <210> 171 <211> 24 <212> DNA <213> artificial sequence <220> <223> GDA1 STM common primer <400> 171 gcatgccctg cccctaagaa ttcg 24 <210> 172 <211> 20 <212> DNA <213> artificial sequence <220> <223> FBP26 L1 Primer <400> 172 tggaggtcag taatcggtcg 20 <210> 173 <211> 38 <212> DNA <213> artificial sequence <220> <223> FBP26 L2 Primer <400> 173 tcactggccg tcgttttacg gattggatgg atgtgaac 38 <210> 174 <211> 40 <212> DNA <213> artificial sequence <220> <223> FBP26 R1 Primer <400> 174 catggtcata gctgtttcct gtccgatgta tgctctggtc 40 <210> 175 <211> 19 <212> DNA <213> artificial sequence <220> <223> FBP26 R2 Primer <400> 175 tgtttctccc cttgtcacc 19 <210> 176 <211> 20 <212> DNA <213> artificial sequence <220> <223> FBP26 SO Primer <400> 176 tggaaatgag ttctcttggg 20 <210> 177 <211> 19 <212> DNA <213> artificial sequence <220> <223> FBP26 PO Primer <400> 177 tcctaaaatc ccgctctgc 19 <210> 178 <211> 21 <212> DNA <213> artificial sequence <220> <223> FBP26 STM Primer <400> 178 actagccccc cctcaccacc t 21 <210> 179 <211> 24 <212> DNA <213> artificial sequence <220> <223> FBP26 STM common primer <400> 179 gcatgccctg cccctaagaa ttcg 24 <210> 180 <211> 20 <212> DNA <213> artificial sequence <220> <223> PSR1 L1 Primer <400> 180 ggcagagaga agtttggtag 20 <210> 181 <211> 38 <212> DNA <213> artificial sequence <220> <223> PSR1 L2 Primer <400> 181 tcactggccg tcgttttact gggcatttct tctactgc 38 <210> 182 <211> 40 <212> DNA <213> artificial sequence <220> <223> PSR1 R1 Primer <400> 182 catggtcata gctgtttcct ggttcgtggt gtccttgatg 40 <210> 183 <211> 19 <212> DNA <213> artificial sequence <220> <223> PSR1 R2 Primer <400> 183 ccttgagccg tttgatgtc 19 <210> 184 <211> 19 <212> DNA <213> artificial sequence <220> <223> PSR1 SO Primer <400> 184 ctgtcattac tcgctcagc 19 <210> 185 <211> 19 <212> DNA <213> artificial sequence <220> <223> PSR1 PO Primer <400> 185 attctgtgtc gctggagtg 19 <210> 186 <211> 24 <212> DNA <213> artificial sequence <220> <223> PSR1 STM Primer <400> 186 cgctacagcc agcgcgcgca agcg 24 <210> 187 <211> 24 <212> DNA <213> artificial sequence <220> <223> PSR1 STM common primer <400> 187 gcatgccctg cccctaagaa ttcg 24 <210> 188 <211> 20 <212> DNA <213> artificial sequence <220> <223> CDC1 L1 Primer <400> 188 aaagggtcgt gtgagaggac 20 <210> 189 <211> 38 <212> DNA <213> artificial sequence <220> <223> CDC1 L2 Primer <400> 189 tcactggccg tcgttttacc tgacgatgct acagatgc 38 <210> 190 <211> 41 <212> DNA <213> artificial sequence <220> <223> CDC1 R1 Primer <400> 190 catggtcata gctgtttcct gtgggcaaga gttggagaga g 41 <210> 191 <211> 21 <212> DNA <213> artificial sequence <220> <223> CDC1 R2 Primer <400> 191 gaacagacag atggggataa c 21 <210> 192 <211> 20 <212> DNA <213> artificial sequence <220> <223> CDC1 SO Primer <400> 192 gctgtgggtg ttgaatgatg 20 <210> 193 <211> 19 <212> DNA <213> artificial sequence <220> <223> CDC1 PO Primer <400> 193 ggcattgacc agcgagttt 19 <210> 194 <211> 20 <212> DNA <213> artificial sequence <220> <223> CDC1 STM Primer <400> 194 cgcccgccct cactatccac 20 <210> 195 <211> 24 <212> DNA <213> artificial sequence <220> <223> CDC1 STM common Primer <400> 195 gcatgccctg cccctaagaa ttcg 24 <210> 196 <211> 20 <212> DNA <213> artificial sequence <220> <223> INP5202 L1 Primer <400> 196 aagagtttgt caccagtgtc 20 <210> 197 <211> 38 <212> DNA <213> artificial sequence <220> <223> INP5202 L2 Primer <400> 197 tcactggccg tcgttttaact ggcaagtaga gagtcacc 38 <210> 198 <211> 40 <212> DNA <213> artificial sequence <220> <223> INP5202 R1 Primer <400> 198 catggtcata gctgtttcct gtgatttggg gcatctggtg 40 <210> 199 <211> 19 <212> DNA <213> artificial sequence <220> <223> INP5202 R2 Primer <400> 199 ggagtttcgc tacattggg 19 <210> 200 <211> 20 <212> DNA <213> artificial sequence <220> <223> INP5202 SO Primer <400> 200 gcattgctac ttcccaaaag 20 <210> 201 <211> 19 <212> DNA <213> artificial sequence <220> <223> INP5202 PO Primer <400> 201 tcttggggatt gggctttgg 19 <210> 202 <211> 24 <212> DNA <213> artificial sequence <220> <223> INP5202 STM Primer <400> 202 ccatagcgat atctacccca atct 24 <210> 203 <211> 24 <212> DNA <213> artificial sequence <220> <223> INP5202 STM common primer <400> 203 gcatgccctg cccctaagaa ttcg 24 <210> 204 <211> 19 <212> DNA <213> artificial sequence <220> <223> YND1 L1 Primer <400> 204 acgcacagtt tgataaccg 19 <210> 205 <211> 38 <212> DNA <213> artificial sequence <220> <223> YND1 L2 Primer <400> 205 tcactggccg tcgttttaca gtggtgtaac ggatggtg 38 <210> 206 <211> 40 <212> DNA <213> artificial sequence <220> <223> YND1 R1 Primer <400> 206 catggtcata gctgtttcct gagtagggcg aatctgagtg 40 <210> 207 <211> 19 <212> DNA <213> artificial sequence <220> <223> YND1 R2 Primer <400> 207 agtgagttct ccgatgtcc 19 <210> 208 <211> 19 <212> DNA <213> artificial sequence <220> <223> YND1 SO Primer <400> 208 gctcgtgatt gggactaac 19 <210> 209 <211> 20 <212> DNA <213> artificial sequence <220> <223> YND1 PO Primer <400> 209 ccatccatct tctccactcc 20 <210> 210 <211> 24 <212> DNA <213> artificial sequence <220> <223> YND1 STM Primer <400> 210 ctccccacat aaagagagct aaac 24 <210> 211 <211> 24 <212> DNA <213> artificial sequence <220> <223> YND1 STM common primer <400> 211 gcatgccctg cccctaagaa ttcg 24 <210> 212 <211> 21 <212> DNA <213> artificial sequence <220> <223> B1026 - M13 Forward extended <400> 212 gtaaaacgac ggccagtgag c 21 <210> 213 <211> 21 <212> DNA <213> artificial sequence <220> <223> B1027 - M13 Reverse extended <400> 213 caggaaacag ctatgaccat g 21 <210> 214 <211> 21 <212> DNA <213> artificial sequence <220> <223> B79 - Screening primer <400> 214 tgtggatgct ggcggaggat a 21 <210> 215 <211> 24 <212> DNA <213> artificial sequence <220> <223> B1454 - NAT split marker primer 1 <400> 215 aaggtgttcc ccgacgacga atcg 24 <210> 216 <211> 24 <212> DNA <213> artificial sequence <220> <223> B1455 - NAT split marker primer 2 <400> 216 aactccgtcg cgagccccat caac 24 <210> 217 <211> 18 <212> DNA <213> artificial sequence <220> <223> B1886 - NEO split marker primer 1 <400> 217 tggaagagat ggatgtgc 18 <210> 218 <211> 18 <212> DNA <213> artificial sequence <220> <223> B1887 - NEO split marker primer 2 <400> 218 attgtctgtt gtgcccag 18 <210> 219 <211> 22 <212> DNA <213> artificial sequence <220> <223> B679 - qRT-PCR primer for ACT1 <400> 219 cgcccttgct ccttcttcta tg 22 <210> 220 <211> 22 <212> DNA <213> artificial sequence <220> <223> B680 - qRT-PCR primer for ACT1 <400> 220 gactcgtcgt attcgctctt cg 22 <210> 221 <211> 19 <212> DNA <213> artificial sequence <220> <223> B8953 - qRT-PCR primer for LAC1 <400> 221 caccctttgg aagttgtgg 19 <210> 222 <211> 19 <212> DNA <213> artificial sequence <220> <223> B8954 - qRT-PCR primer for LAC1 <400> 222 tgataattgc agagtaccg 19 <210> 223 <211> 19 <212> DNA <213> artificial sequence <220> <223> B3737 - qRT-PCR primer for BZP4 <400> 223 agccaggtaa tcttggagg 19 <210> 224 <211> 19 <212> DNA <213> artificial sequence <220> <223> B8521 - qRT-PCR primer for BZP4 <400> 224 caatatacga atcactccc 19 <210> 225 <211> 20 <212> DNA <213> artificial sequence <220> <223> B6394 - qRT-PCR primer for HOB1 <400> 225 cctcgcaagt tccccagcta 20 <210> 226 <211> 19 <212> DNA <213> artificial sequence <220> <223> B8519 - qRT-PCR primer for HOB1 <400> 226 gtatgaggtc ttgtccacc 19 <210> 227 <211> 20 <212> DNA <213> artificial sequence <220> <223> B8654 - qRT-PCR primer for ITR1A <400> 227 cttcaaccga ggtcatactc 20 <210> 228 <211> 19 <212> DNA <213> artificial sequence <220> <223> B8655 - qRT-PCR primer for ITR1A <400> 228 agattccgat accaagggc 19 <210> 229 <211> 20 <212> DNA <213> artificial sequence <220> <223> B8658 - qRT-PCR primer for ITR3C <400> 229 ccctttggtc aggtgatttc 20 <210> 230 <211> 21 <212> DNA <213> artificial sequence <220> <223> B8659 - qRT-PCR primer for ITR3C <400> 230 gctgaaatag ggatggaaca g 21 <210> 231 <211> 20 <212> DNA <213> artificial sequence <220> <223> B8656 - qRT-PCR primer for MPR1 <400> 231 cgaggttctt gatgatgctg 20 <210> 232 <211> 20 <212> DNA <213> artificial sequence <220> <223> B8657 - qRT-PCR primer for MPR1 <400> 232 atccgaggaa agtctgagcc 20 <210> 233 <211> 19 <212> DNA <213> artificial sequence <220> <223> B8598 - qRT-PCR primer for FZC31 <400> 233 aaatgtcccg aaaaggaag 19 <210> 234 <211> 21 <212> DNA <213> artificial sequence <220> <223> B4951 - qRT-PCR primer for FZC31 <400> 234 tctcttcttc ttctgacctg c 21 <210> 235 <211> 17 <212> DNA <213> artificial sequence <220> <223> B9243 - qRT-PCR primer for GAT201 <400> 235 catcccgtcg ccacagc 17 <210> 236 <211> 19 <212> DNA <213> artificial sequence <220> <223> B9422 - qRT-PCR primer for GAT201 <400> 236 ggaggtatggc tgaaatctg 19 <210> 237 <211> 20 <212> DNA <213> artificial sequence <220> <223> B9061 - qRT-PCR primer for PDR802 <400> 237 tttcgtagcc tgtaagtggc 20 <210> 238 <211> 19 <212> DNA <213> artificial sequence <220> <223> B4913 - qRT-PCR primer for PDR802 <400> 238 ggaacattgg gaaaaggtg 19

Claims (4)

Contacting a candidate material with Cryptococcus neoformans;
PHS1, a gene expressing a phosphatase associated with the cell membrane stress sensitivity of Cryptococcus neoformans; and mRNA of one or more genes selected from the group consisting of VPS29, YMR1, SSU72, NEM1, YVH1, SIT4, GUA1, INP5201, DBR1, SIW14, SDP102, OCA1, GDA1, FBP26, PSR1, CDC1, INP5202, and YND1, or Measuring the expression level of the expressed protein or measuring the activity of the protein; and
An antifungal agent screening method for Cryptococcus neoformans comprising the step of determining that the candidate material has antifungal activity when the measured expression level or activity is significantly reduced compared to a control group not contacted with the candidate material,
The cell membrane stress sensitivity is an antifungal agent screening method in which the growth of Cryptococcus neoformans is inhibited in a medium to which a cell membrane stress inducing substance is added. delete According to claim 1,
The mRNA expression level is measured by reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction, real-time reverse transcription polymerase reaction, RNase protection assay (RPA), Northern blotting, DNA chip (Microarray) or RNA sequencing analysis. to perform,
Antifungal agent screening method. According to claim 1,
The expression level of the protein is measured by Western blot, ELISA (enzyme linked immunosorbent assay), radioimmunoassay (RIA), radioimmunodiffusion, tissue immunostaining, immunoprecipitation assay, complement fixation assay ( Complement Fixation Assay), flow cytometry (Fluorescence Activated Cell Sorter, FACS), or protein chip (protein chip),
Antifungal agent screening method.

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