CN1189537A - Component of white monilia with reverse transcription transposons structure - Google Patents

Abstract

A leucoidiomycetic component Tcal-2 with reverse transcription transposons structure is composed of 5624 nucleotides and can be transcripted into 5.5kb-long RNA. The output of transcription is controlled by temp, that is more at 25 deg. C than at 37 deg.C. The Tcal-2 component comprises two long-terminal repeated sequences Tcal-2LTR composed of 388 nucleotides. The clone of Tcal-2 and Tcal-2LTR provides useful molecular tool for classifying leucoidiomycetic spawns and also lays a foundation for screen antifungus medicines.

Description

Candida albicans has the element of retrotransposition minor structure

The invention belongs to the retrotransposon field, especially address and from Candida albicans, be cloned into an elements T ca1-2 with retrotransposition minor structure.

Retrotransposon is the mobile gene that a class and retrovirus have the common structure feature, extensively is present in the eukaryote.Retrovirus is to cause the one of the main reasons that the animals and human beings class forms tumour, retrovirus is by influencing the numerous expression of gene regulation and control of host cell with genomic integration and reorganization, and retrovirus can be used as the cell lineage mark and chromosome marker is applied to Mammals auxology and genetic research.Recombinant retrovirus is a good carrier in addition, has been successfully used to gene therapy.Found in the cereuisiae fermentum five class Ty elements (Ty1, Ty2, Ty3, Ty4, Ty5) their swivel base and reorganization have caused genomic rearrangement, activate or the expression of deactivation transposon contiguous gene.The Ty element is used to clone the gene with new function as a useful molecular tool.As expression vector, swivel base is inserted in the genome the Ty element, constitutes endogenic expression.Retrotransposon is dispersed in the seed that can become evolution in the genome, forms the new gene or the structural domain of gene by sudden change, and the swivel base of retrotransposon can promote genomic flowing, and helps the multifarious development of biological heredity.Therefore the research of retrotransposon helps illustrating of gene expression regulation mechanism, for the molecular evolution Mechanism Study of cell provides new approaches, as the expression system of antigenic determinant, is a kind of important instrument in the genetically engineered.

In more than the 100 kind of candidiasis, the pathogenecity of Candida albicans is the strongest, can cause the moniliosis of tissue surface and depths.Candida albicans opportunistic ground causes stomatitis, vaginitis and various skin inflammation, can infect respiratory tract, enteron aisle, also can cause intracardiac Valvulitis, also relates to histoorgans such as liver, brain, and severe infections has lethal possibility.Candida albicans is one of human body endosymbiontic microorganism flora, often exists with thalline and mycelia form, also forms pseudohypha sometimes.Candida albicans does not have sex to change, and karyomit(e) is diploid, and hyphoid Candida albicans has stronger infectivity and pathogenecity (Odds, F.C., 1988, Candida andcandidosis.A review and bibliography., Bailliere Tindal, London.).Multiple ambient conditions such as pH, temperature, serum, N-acetylglucosamine etc. can cause that all the form of Candida albicans changes, and form changes with it pathogenicly has certain related (Mattia, an E., G.Carruba, 1982, J.Bacteriol., 152:555-562.; Buffo, J., M.A.Herman, 1984, Mycopathologia, 85:21-30.).Phenotype and genotypic difference between the different Candida albicans bacterial classifications are bigger, and pathogenecity is also different with the toxicity performance, and Candida albicans also usually shows stronger toxicity under 25 ℃ of conditions.The pathogenesis of research Candida albicans, toxicity performance and form generation regulatory mechanism are one of hot research in recent years (Saporito-Irwin, S.M., C.E.Birse, 1995, Mol.Cell.Biol., 15:601-613. on molecular level; Hoyer, L.L., S.Scherer, 1995, Mol.Microbiol., 15:39-54.; Matthews, G.D., 1997, J.Bacteriol., 179:7118-7128).So far because the shortage of special-purpose antifungal drug makes many trouble diabetes, leukemia, malignant tumour, especially the patient of immunological competence decline (as the AIDS disease) is behind fungal infection, and no medicine can be executed, and life is constituted very big threat.The research of retrotransposition sub-element regulation rule will help the screening of antifungal drug.Has important biology medical significance.

The object of the invention provides the elements T ca1-2 full length nucleotide sequence that a kind of Candida albicans has the retrotransposition minor structure, and this element can be used for the classification of Candida albicans bacterial classification to be identified.The research of this element regulation rule will help the screening of antifungal drug.

The 500bp α tumor-necrosis factor glycoproteins (Chen of the present invention before to be separated to, J.and W.A.Fonzi, 1992, J.Bacteriol, 174:5624-5632.) be probe, utilize Southern hybridization analysis and molecule clone technology to be cloned into two elements T ca1-1 and Tca1-2 with retrotransposition minor structure from the chromosomal DNA of Candida albicans SC5314 bacterium, the physical map of Tca1-2 in karyomit(e) seen Fig. 1.Tca1-2 element full length nucleotide sequence is 5624bp (Fig. 2).The Tca1-1 componentry nucleotide sequence of having measured shows that the nucleotide sequence homology of Tca1-1 element and Tca1-2 element is very high, and size is close, and physical map is similar, and they all have the constitutional features of retrotransposon.Two identical long terminal repeat LTR that arrange are in the same way all contained at Tca1-1 element and Tca1-2 element two ends separately, the LTR at the LTR at Tca1-1 two ends and Tca1-2 two ends is made up of 388 Nucleotide, and LTR has the constitutional features of other retrotransposon LTR.Have only 4 Nucleotide different (Fig. 3) between the nucleotide sequence of Tca1-1LTR and Tca1-2LTR.Through database retrieval, the nucleotide sequence of Tca1-2 element has only two LTR at two ends and the α tumor-necrosis factor glycoproteins homology that the inventor has delivered, and all the other 4848 nucleotide sequences are not found homologous sequence.Tca1-1 and Tca1-2 element can be transcribed out the long transcription product of 5.5kb respectively, and transcribing of they all is subjected to temperature regulation, the Northern hybridization analysis.The transcription product that shows 25 ℃ is more than 37 ℃ transcription product (Fig. 4).With strand Tca1-2 DNA is probe, and the transcriptional orientation of determining Tca1-2 is to transcribe right LTR (Fig. 1) from left LTR.With inboard sequence of Tca1-2 element and Tca1-2LTR sequence is that probe carries out the Southern hybridization analysis to tens kinds from the clinical pathogenic albicans strain that is separated to, and these Candida albicans bacterial classifications can be divided into some groups, exists the intrinsic contact between bacterial classification.Results of hybridization show from the difference between different countries and the geographic Candida albicans greater than from identical geographic bacterial strain (Fig. 5, Fig. 6).The Tca1-2 element can be used for the classification of Candida albicans bacterial classification to be identified.

Advantage of the present invention:

1. the nucleotide sequence of a brand-new Tca1-2 element of Candida albicans is provided, and it has the constitutional features of retrotransposon, can develop into a kind of molecular tool, is applied to genetically engineered.

2.Tca1-2 transcribing of element is subjected to temperature adjusting, helps to illustrate the pathogenesis of Candida albicans, might be used for the screening of antifungal drug.

Identify 3.Tca1-2 can be used for the classification of medical science pathomycete Candida albicans bacterial classification with the Tca1-2LTR sequence, be applied to oidiomycotic treatment.

The present invention is further elaborated by the following drawings and embodiment, but does not limit the scope of the invention.

Description of drawings:

Physical map and the sequence signature of Fig. 1 Tca1-2 element in karyomit(e).

Long terminal repeat LTR goes out with the square frame frame among the figure, the transcriptional orientation of Tca1-2 element from left to right as shown by arrows, the 1kb scale is as shown in the figure.

The nucleotide sequence of Fig. 2 Tca1-2 element.

The comparison of Fig. 3 Tca1-1LTR and Tca1-2LTR sequence.

The Northern hybridization analysis of Fig. 4 Tca1-1 and Tca1-2 element

The total RNA of Candida albicans SC5314 uses inboard sequence probe of (A) Tca1-1 and (B) the inboard sequence probe hybridization of Tca1-2 respectively among the figure.Same RNA dyes with EB respectively.

The Southern hybridization analysis of Fig. 5 Tca1-2 element and Tca1-2LTR sequence and Candida albicans chromosomal DNA.

Among the figure with (A) Tca1-2LTR sequence and (B) the inboard sequence of Tca1-2 element be probe, with the chromosomal DNA hybridization of Candida albicans EcoRI enzymolysis.Bacterial classification 1 to 30 from the U.S..

The Southern hybridization analysis of Fig. 6 Tca1-2 element and Tca1-2LTR sequence and Candida albicans chromosomal DNA

Among the figure with (A) Tca1-2LTR sequence and (B) the inboard sequence of Tca1-2 element be probe, with the chromosomal DNA hybridization of Candida albicans EcoRI enzymolysis.Bacterial classification 1 is from the U.S., bacterial classification 31 to 37 from China.

Embodiment 1

1, the preparation of probe

With restriction enzyme StyI and KpnI (available from U.S. BRL company) enzymolysis α tumor-necrosis factor glycoproteins (Chen, J.and W.A.Fonzi, 1992, J.Bacteriol, 174:5624-5632.), reaction conditions carries out to specifications, separates 300bp StyI-KpnI dna fragmentation, use the 32P mark, be referred to as the α probe.Isotopic compound [α-32P] dATP and probe mark test kit are available from Amersham company, and operation steps is carried out to specifications.With separate the dna fragmentation that Tca1-1 and Tca1-2 element inboard do not contain the LTR long terminal repeat respectively with quadrat method, use the 32P mark, be referred to as the inboard sequence probe of Tca1-1 and Tca1-2.Separate the dna fragmentation of 388bp in the Tca1-2LTR sequence, use the 32P mark, be referred to as the Tca1-2LTR probe.

2, the clone of Tca1-1 and Tca1-2 element

Mix mutually with KW251 bacterium liquid (this laboratory has) in Candida albicans SC5314-λ GEM-12 chromosomal DNA storehouse (being given by professor W.A.Fonzi of U.S. George Washington University), the agar that adds the 3ml fusing then, spread to the LB plate, cultivated 10～12 hours for 37 ℃.Phage DNA is transferred to the nylon Hybond membrane, uses the α probe hybridization, and (quick hybridization liquid is available from Amersham company, and operation steps is carried out to specifications) obtains 68 positive colonies.Cut evaluation through enzyme, clone λ CJY-3 and clone λ CJY-4 contain Tca1-1 and Tca1-2 element respectively, and extracting λ CJY-3 and λ CJY-4DNA connect the BamHI site that is inserted into pBSK (+) plasmid with BamHI restriction enzyme enzymolysis.PBSK (+) plasmid is available from STRATAGENE company, and the T4 dna ligase is available from U.S. BRL company.The subclone recombinant plasmid that obtains is used multiple restriction enzyme enzymolysis respectively, makes physical map, and the physical map of Tca1-2 element in karyomit(e) seen Fig. 1.

Embodiment 2

1, the nucleotide sequencing of Tca1-2 element

According to the physical map of Tca1-2 element, the dna fragmentation of Tca1-2 element is distinguished subclone in M13mp18 and M13mp19 carrier, M13mp18 and M13mp19 carrier are available from Amersham company.Measure dna sequence dna, the determined dna sequence test kit is available from U.S. USB company, and the operation by specification carries out.Record Tca1-2 element nucleotide sequence, total length is 5624bp, and Tca1-2 element full length nucleotide sequence as shown in Figure 2.

2, sequential analysis and comparison

The analysis and the homology comparison query of Tca1-2 element nucleotide sequence are finished by the NCBI E-mail service, and the storehouse of retrieval comprises GenBank, EMBL, GenPept, Swissprot, PIR, DBEST etc.Sequential analysis shows that the Tca1-2 element is a new element with retrotransposition minor structure.5 ends of Tca1-2 element and 3 ends comprise the long terminal repeat LTR of a 388bp respectively, two LTR are with equidirectional arrangement, right LTR inboard have the normal chain primer binding site (+PBS), there is the minus strand primer binding site LTR inboard, a left side (PBS), there is the direct repeat of 5bp in two LTR outsides, and the constitutional features of Tca1-2 element in karyomit(e) as shown in Figure 1.

The transcriptional control of embodiment 3 Tca1-1 and Tca1-2 element

Candida albicans SC5314 (Chen, J.and W.A.Fonzi, 1992, J.Bacteriol, 174:5624-5632.) always preparation, electrophoresis, the commentaries on classics film of RNA all carry out (Sambrook, J. by the method that " molecular cloning, experiment guide " introduced, Fritsch, E.F., and Maniantis, T.Molecular Cloning, Cold SpringHarbor Laboratory Press, 1989).Total RNA placed 10 minutes in room temperature after transferring to nylon membrane, crosslinked with 0.7J/cm2 on ultraviolet lamp, pack into after soaking with 2XSSC in the hybrid pipe, add the quick hybridization liquid (available from Amersham company) of 5ml in 68 ℃ of preheatings, in 68 ℃ of prehybridizations 30 minutes, add respectively behind the inboard sequence probe of Tca1-1 of sex change and the inboard sequence probe of Tca1-2 again in 68 ℃ of hybridization 2 hours.Wash film, film is placed on the filter paper dry, radioautograph, the result is as shown in Figure 4.The Northem results of hybridization shows that the Tca1-1 element can synthesize the long transcription product (A among Fig. 4) of 5.5kb, the Tca1-2 element also can synthesize the long transcription product (B among Fig. 4) of 5.5kb, the transcription product size of Tca1-1 element and Tca1-2 element is close, transcribing of they all is subjected to temperature adjusting, and 25 ℃ transcription product is more than 37 ℃ transcription product.The transcriptional orientation of Tca1-2 element is to transcribe right LTR (indicating) from left LTR among Fig. 1.

Embodiment 4 Tca1-2 elements and Tca1-2LTR sequence are used for the classification of Candida albicans bacterial classification and identify

1 to No. 37 Candida albicans bacterial classification chromosomal DNA of extracting, No. 1 to 30, bacterial classification be from the U.S. (being given by professor W.A.Fonzi of U.S. George Washington University), and bacterial classification 31 to 37 is from China (being given by professor Wu Shaoxi of institute of internal medicine of Chinese medical courses in general institute).Chromosomal DNA restriction enzyme EcoRI enzymolysis, electrophoresis, commentaries on classics film.The extracting of chromosomal DNA, enzymolysis, electrophoresis, commentaries on classics film carry out (Sambrook, J., Fritsch by the method that " molecular cloning experiment guide " introduced, E.F., and Maniantis, T.MolecularCloning, Cold Spring Harbor Laboratory Press, 1989).Nylon membrane after shifting with crosslinked with 0.7J/cm2 on the ultraviolet lamp, is added quick hybridization liquid (available from Amersham company), in 42 ℃ of prehybridizations 60 minutes.Add respectively behind the inboard sequence probe of Tca1-2 of sex change and the Tca1-2LTR probe again in 42 ℃ of hybridization 4 hours.Wash film, radioautograph, result such as Fig. 5, shown in Figure 6.Tca1-2LTR probe hybridization result shows that the Candida albicans bacterial classification can be divided into some groups, from the genetic correlation of areal bacterial classification recently from the genetic correlation big (A among Fig. 5, the A among Fig. 6) of different areas.The inboard sequence probe hybridization of Tca1-2 result supports the classification qualification result of Tca1-2LTR sequence shown in B and the B among Fig. 6 among Fig. 5.Show that 1 to No. 3 Candida albicans belongs to a group, 4 to No. 9 and No. 14, No. 31 Candida albicanss belong to a group, 10 to No. 13 and No. 32 Candida albicanss belong to a group, 15 to No. 17 Candida albicans belongs to a group, 18 to No. 21 Candida albicans belongs to a group, 22 to No. 24 and 26 to No. 28 Candida albicanss belong to a group, and 25,29, No. 30 Candida albicans belongs to a group, and 34 to No. 37 Candida albicans belongs to a group.

Claims (2)

1. A method Candida albicans retrotransposon structure elements Tca1-2, characterized in that It has the following DNA nucleotide sequence: ATTGCTGTTCGCTATAGAGAGATTTCCTAGCCGGAATGCACGACAATCCTGAGACGGAAG 60 TCGATCGTCGATGCCCATGGTGCGTGGTGAAAAATTTTCTTAGAAAATTTGTTCTTTCCT 120 TCAACTGCTTTTAAGAAAGAGAGGTTCAAGTGGTTTAAGTACGACGGTCACAAAGATTGC 180 GGCTTATGAGGCCCGAACTGAGTTGAAATACAAAATCAAGATATAATTATATACCTTACT 240 TGTCCATATTGTTTTATAATACATTCTTCAGATATTTAAATTTCTGTGTATCAACCTATA 300 AAACAGAGATACATTCAGTGCATTTAGTATACTGAGTGAACTGGTACCTGTGACATTCAA 360 GATAACTGTTTCGCGCACGCTGGCAGACGAACAGATTAGAAGCTTGGTAAAGTTCTGCTT 420 TGCTCAATAGGTTTCAGATTCAGAAAGATTGTTAAAACTTAGATCATCTTCGTTCATCAC 480 AAACCAAGAACTTTACGGAATGTACGAATATCACTTTCATTAGTAGATAATTCGTTACTT 540 AATCCAGTGATTAATCTTGAGGTTCGAAAGATGGTTAATAGAAATTTATTTGACAATTAC 600 GACTAAGGTTACATAATAAATCATTGGTATCACGGCTATGAAAGCTTCCAAGATGTGATT 660 TTAATAACAGAGTGTTTTTGGTCTCAACAGATGAGAATACATTGAATTTAATGAATTGCT 720 AACAAAAGTCATCAATTAGTCTACGACTGAAATGGAATACTATTCATATTGTGTTAATGA 780 TTGACATTAAAAATGAACAAAATAAAAGGTATCATAGATTTTATGCTATTCAAAGTGATG 840 GTGGTGTTGACAAAACGTTTTGAAAAGTAACGATATTTGGACTAAAAAGGATTCAAGAAT 900 TATGTCATTGCTGAATAATTTTGGAGAACCTGCCTTAATAATTTTATGAGGTGTCTACTA 960 TATGGTAAAGTTTTGATGAACAAAAGAAATGAAATTATAATGATTGCCAAATTTGTGCAC 1020 ATTCCCATTAAAGGGATAAGAGTCTACAATAAAGCAACAGTAGTTTTCATGAACACTACT 1080 Follow-up TAAACACCAAGGAAGAACAAGATCTATGAGGTGCCATTTTTAAAAAATTAAGTTGAAGGG 1140 GAAAATCTAAAAGTACCTATCAATTTATGGAAGTTTATTTGTTAATCTGTCAACTATGTG 1200 AAGGAACGTATAGTCGAACTACTTGGTAACTCATTTCGTTAATCCACACATGAACGTTTA 1260 AAACCTAAAAAATGAAGTCATACATCTTTTCATACAATGATCTTATCAATTCAAGACATA 1320 CCTTTGTGATGTTATAATTTTGTAAGTCATTCAAAGGGGAGATTTGCACTGAATAACTCA 1380 TCTATGCTCATACATGCTGGTATTGAAAAATTGATTTTCACGTTTGTTACCTTTCAGAAG 1440 TCTATGCATTACAATAGTGAAAATTCAATGCGAACCCATTGCTAAATTTACCAGACTCAA 1500 AAGAGAAAATGATTGAGATAAAAAATTACAGAGATTATTCACAAATCGTCCAGTATTGTT 1560 AAGAGAAAAGTGAATTAGATGATAATCATAAGCATAGGAATCAACTTCATGATGTCAGAT 1620 AAACCCATTATGGTATTTTATCTATCATTATTCCAACATGATATCCCAGAATACATGTGA 1680 TAATGAAATTCAATAAACTGGTTAAAGAGAAATTTTGAAATATGGCTTCTTTAAGAAATT 1740 TTATAAAATGAAAAGAGTTGCTGAAATCCATGCTATTACACACTTTTTCATATTCCTGAG 1800 AAGTGAAAGCTACGTCACACAGTCTTCGTTAGATAAGAACTCTAAATGTTGGAATATTTG 1860 TACCAGGACATGCATAAGCATCCAGTCACAATGGCCATAAACATGAGAAACCTACCCAAT 1920 GAAGACTACCAATGAATTATATACTGAAAAAATGTCCAAGATATGAGTATTAATTAACTA 1980 TTCACCTTATGAATATCCCAAATATTCAGACCTATCATAATGATTATTTCATAGACAAAA 2040 ATGAGTACCAATTCCACATTATGAGTTGTTGAATGTTGTGGAGTATGAAATTATGATGAA 2100 CATCCACGTATAAATTAGTTGTCGAGAATTATCGATCGAACAGATATTAGACCTAGAGCT 2160 GATCCCACCTGGCAACCGTACCTGATGCCGTCATACATCAAGTACACCATACTGTACAGA 2220 CACCTGATCATGGGGTGTAAAGATACCATGATCAATCACACCGACTATTACGATCTGGGG 2280 AGGGTAATTACCCCGGACACCAGGTGCGCACCGAAAAATTGGGAATTCGAGATCGCGGGC 2340 CTACCACCCTAAACACTGCGATTTGATGTTAGGTGTAAACGACGAAACATACGATACCGT 2400 GATACATCGAGGAATCATATTGGTTCTCTAGATTCCAAGATGATTGCCACACCATCTTTA 2460 Follow-up CCAAAACGAATAGATAATCAAAATGATATCAATTCAGGGAAGATGACGTTACTGCAAAGT 2520 TTCTTAAAGCAGGTAACGAACTGAAATTATATAAGACATTGTCATAAAGCAAAACAACAT 2580 GTACATGACATTGCCGCAAACATTGACTTATGCATAAGAAATACATCAGATGAGATAAGA 2640 AACTGAAAAAGTAATCTCATTTTTTCTACGGCAAATTTTACAAAGAAGACAATATGGGAA 2700 TAAAATAGACCATATTACCAAATTATGAAATGGACACTCTCTAAAGACTTATCAATCATT 2760 ACAAGCTGCTTTTAACGTGGAATTTTCTAAGTGTTCGACATTGATGTTGTTTTTATCACT 2820 ATCATCATATTTCCAAAGTTTTCAGGGTCGATGATTATAGAACAAGAGCTAACCTCAACG 2880 AGAGCACTGTTTGAAAGCACCCGACACCCCGAGGATTCAATAAAATTGAGGATAGAACGG 2940 TAAAATAGTAATCACTAACGAAGACATGCTAGAAATTTTTACATGAAAGACATTACATTT 3000 GCCCCTATGTTCAATGAACACGAATGAAATTTTGGATATACAACCACATTTTAAGTAATA 3060 GGTTATGGGTATAAAACTAAACTATAGCTAGTTGGTATTGGTAAAGGAGAAAGGTCCAAT 3120 GACATGATTTTGTCAATAAGTGCTTCAAGAAAATAATATTAGCCAACGGATAAGATTGAT 3180 CTAGACAGGAGCAATAACAACACGATAACACTGATAAATTGCAAGAACCAAGTAGGTCTC 3240 AATATTTGATGGTTATTATCACTCTTTCCCCAACAAAAGAATACAAATGTTCAAACCAAC 3300 ACCACTTCAGGGGAGGAGAATTTATGAATAAAATTGATATGAAGTAGTGGATGATTTGGA 3360 AAAAAATTCGGTAATGTTATGAATTTACCAATAGAATAAGATAACATTACTACCCCTGAT 3420 TGGTATATTAGACAGAACTTGTGATGATACTAAAACTCGAAAAGAGATGTTTCAAGCAGG 3480 GGCATATAATGTGCAAAATGTCACAATGACTCTAAGATTCAAATGGCATCATGATTTCTG 3540 AACTAAATACCAGCCAAACACTCTCGGTTGATACGTCCAATTTAAAAGACGGTCACCGAA 3600 AACAAAGTGAAAGTAAGTGAAATCATATGAATAGATTCACAAGAAATCGTAATGGAAACA 3660 CAATAATCATTTCATAAATCTTTGAAGTATACACAAGAATTGAGGTCAATAATTATCTAT 3720 TAAAAGACTACCACAAGGTTGACTTTATGATCCCAATACACCAATAGAGGAGATTGAAAC 3780 Follow-up ATCCTAGGTCAACGTCGAAGATGGAAATTTGAGACAAAGATGAATAAAAGGTCGACCGAT 3840 AACATTGTTTGATGAATCCAAAGAGAATTGGTACTTCAATACATTAAATAAAGATCTTCC 3900 GTTAAATGTAAGATACGAAATTGATGTGAATCCGTTACTTAAATATCTAAAGAAGAATAA 3960 TGCAACTTATGAGATATGTTGAAAACATGAATTAGCTTAAATCACACTAAAGTGAGAAGT 4020 TTGATTTTAAGAAACATACCATAGAGTATTTATAATATCGTCCATCCTACTGCAAAAAGA 4080 CTATTCCATGCTATACTCAAATCAAAAAACTGGGTACATACTTAATGATGATGACTAAAT 4140 TCAAACCAAGTGTCACCATTTCTAGATGGCTACTCAAAAATGAGAATGATGGCAAACAAA 200 TGGAAGATAGATCAATGTTTGAATTGCTTAGACAGAGATTTTGGCTAAAACACCAGCAGA 4260 CTATGAATGCGGAATACAAGTGATAAAGTTTTACAACTTCACCAAATTGTGAATATAATC 4320 AATCTGGAATCCCGGAAATAGAAATCAAAGATTGAAAATTTTTCGAAATCGACTAATAAA 4380 ATGCATGATTTCGTCAAATTTAAACAACACATACGCAAATATATTAAATTAGAATACCGA 4440 TGTGGATTGAAAATGGAGGTGAATACGTGTAAGTACGGTTAAAATTCCACAGTTTTAATT 4500 GTGTTTCATGGCATGTGTCAAGATTATCCATTTGCATAAACTTATCGAAATATTGGTAGT 4560 AAGGAAACAAGGTGAATGCCGGCCGGATTTACCAAGTACTGAAACTCATATTAGGTTTGG 4620 ATGTATGAAAATCGAGATATCTGATAACTACAGAAAACACATATACTACTTAAAAAATAC 4680 CCGAAGTCATTTTCATATTACAATCGGATAGTTAAACAAGGGGAGCTAACGCTCAAAGAG 4740 AGAGTCACATATTTGAATATGACTGTTGACACAGATCTTTATCTAGTGCGGGTTCATAGA 4800 TCTAGTGCTAAGAAAATAGATTGAGTTAATATGATCATTAGTGAATGAAGAAATGATAAC 4860 GTGAAGCGATGTTACAAGAAGATTACGATACAAAAGTGATACTTACGTTAAAATTATTCA 4920 GTATTTGCAAGCATATTAGATTAATAAGGTTAAAATTAACAAAAAACAATTAGGTCCATA 4980 AGCCCATATTACGAAATTATTCTTAGAAAAAGTTATGATTAATTTTCAATCCAACATTAC 5040 AATATTATCTCAAAATAAAGATGCTAGGTTAAAACACACTATTGTCCTTAGAGAAATGGA 5100 AGTTATGTGGAGTTAAAAAATAATCAGTGGAAAGAATCACCGACACAATGATTTGACATC 5160 Follow-up AATTGTTGATAACAAAGTCGCTAGAAGAGATTAAGGAATCAATGATGAACCATGATTACT 5220 GAATCAGGGAGTGTTCGCTATAGAGAGATTTCCTAGCCGGAATGCACGACAATCCTGAGA 5280 CGGAAGTCGATCGTCGATGCCCATGGTGCGTGGTGAAAAATTTTCTTAGAAAATTTGTTC 5340 TTTCCTTCAACTGCTTTTAAGAAAGAGAGGTTCAAGTGGTTTAAGTACGACGGTCACAAA 5400 GATTGCGGCTTATGAGGCCCGAACTGAGTTGAAATACAAAATCAAGATATAATTATATAC 5460 CTTACTTGTCCATATTGTTTTATAATACATTCTTCAGATATTTAAATTTCTGTGTATCAA 5520 CCTATAAAACAGAGATACATTCAGTGCATTTAGTATACTGAGTGAACTGGTACCTGTGAC 5580 ATTCAAGATAACTGTTTCGCGCACGCTGGCAGACGAACAATTGC 5624 ...
2. A method Candida albicans retrotransposon structure elements Tca1-2, characterized in that It has the following DNA nucleotide sequence: ATTGCTGTTCGCTATAGAGAGATTTCCTAGCCGGAATGCACGACAATCCTGAGACGGAAG 60 TCGATCGTCGATGCCCATGGTGCGTGGTGAAAAATTTTCTTAGAAAATTTGTTCTTTCCT 120 TCAACTGCTTTTAAGAAAGAGAGGTTCAAGTGGTTTAAGTACGACGGTCACAAAGATTGC 180 GGCTTATGAGGCCCGAACTGAGTTGAAATACAAAATCAAGATATAATTATATACCTTACT 240 TGTCCATATTGTTTTATAATACATTCTTCAGATATTTAAATTTCTGTGTATCAACCTATA 300 AAACAGAGATACATTCAGTGCATTTAGTATACTGAGTGAACTGGTACCTGTGACATTCAA 360 GATAACTGTTTCGCGCACGCTGGCAGACGAACAGATTAGAAGCTTGGTAAAGTTCTGCTT 420 TGCTCAATAGGTTTCAGATTCAGAAAGATTGTTAAAACTTAGATCATCTTCGTTCATCAC 480 AAACCAAGAACTTTACGGAATGTACGAATATCACTTTCATTAGTAGATAATTCGTTACTT 540 AATCCAGTGATTAATCTTGAGGTTCGAAAGATGGTTAATAGAAATTTATTTGACAATTAC 600 GACTAAGGTTACATAATAAATCATTGGTATCACGGCTATGAAAGCTTCCAAGATGTGATT 660 TTAATAACAGAGTGTTTTTGGTCTCAACAGATGAGAATACATTGAATTTAATGAATTGCT 720 AACAAAAGTCATCAATTAGTCTACGACTGAAATGGAATACTATTCATATTGTGTTAATGA 780 TTGACATTAAAAATGAACAAAATAAAAGGTATCATAGATTTTATGCTATTCAAAGTGATG 840 GTGGTGTTGACAAAACGTTTTGAAAAGTAACGATATTTGGACTAAAAAGGATTCAAGAAT 900 TATGTCATTGCTGAATAATTTTGGAGAACCTGCCTTAATAATTTTATGAGGTGTCTACTA 960 TATGGTAAAGTTTTGATGAACAAAAGAAATGAAATTATAATGATTGCCAAATTTGTGCAC 1020 ATTCCCATTAAAGGGATAAGAGTCTACAATAAAGCAACAGTAGTTTTCATGAACACTACT 1080 Follow-up TAAACACCAAGGAAGAACAAGATCTATGAGGTGCCATTTTTAAAAAATTAAGTTGAAGGG 1140 GAAAATCTAAAAGTACCTATCAATTTATGGAAGTTTATTTGTTAATCTGTCAACTATGTG 1200 AAGGAACGTATAGTCGAACTACTTGGTAACTCATTTCGTTAATCCACACATGAACGTTTA 1260 AAACCTAAAAAATGAAGTCATACATCTTTTCATACAATGATCTTATCAATTCAAGACATA 1320 CCTTTGTGATGTTATAATTTTGTAAGTCATTCAAAGGGGAGATTTGCACTGAATAACTCA 1380 TCTATGCTCATACATGCTGGTATTGAAAAATTGATTTTCACGTTTGTTACCTTTCAGAAG 1440 TCTATGCATTACAATAGTGAAAATTCAATGCGAACCCATTGCTAAATTTACCAGACTCAA 1500 AAGAGAAAATGATTGAGATAAAAAATTACAGAGATTATTCACAAATCGTCCAGTATTGTT 1560 AAGAGAAAAGTGAATTAGATGATAATCATAAGCATAGGAATCAACTTCATGATGTCAGAT 1620 AAACCCATTATGGTATTTTATCTATCATTATTCCAACATGATATCCCAGAATACATGTGA 1680 TAATGAAATTCAATAAACTGGTTAAAGAGAAATTTTGAAATATGGCTTCTTTAAGAAATT 1740 TTATAAAATGAAAAGAGTTGCTGAAATCCATGCTATTACACACTTTTTCATATTCCTGAG 1800 AAGTGAAAGCTACGTCACACAGTCTTCGTTAGATAAGAACTCTAAATGTTGGAATATTTG 1860 TACCAGGACATGCATAAGCATCCAGTCACAATGGCCATAAACATGAGAAACCTACCCAAT 1920 GAAGACTACCAATGAATTATATACTGAAAAAATGTCCAAGATATGAGTATTAATTAACTA 1980 TTCACCTTATGAATATCCCAAATATTCAGACCTATCATAATGATTATTTCATAGACAAAA 2040 ATGAGTACCAATTCCACATTATGAGTTGTTGAATGTTGTGGAGTATGAAATTATGATGAA 2100 CATCCACGTATAAATTAGTTGTCGAGAATTATCGATCGAACAGATATTAGACCTAGAGCT 2160 GATCCCACCTGGCAACCGTACCTGATGCCGTCATACATCAAGTACACCATACTGTACAGA 2220 CACCTGATCATGGGGTGTAAAGATACCATGATCAATCACACCGACTATTACGATCTGGGG 2280 AGGGTAATTACCCCGGACACCAGGTGCGCACCGAAAAATTGGGAATTCGAGATCGCGGGC 2340 CTACCACCCTAAACACTGCGATTTGATGTTAGGTGTAAACGACGAAACATACGATACCGT 2400 GATACATCGAGGAATCATATTGGTTCTCTAGATTCCAAGATGATTGCCACACCATCTTTA 2460 Follow-up CCAAAACGAATAGATAATCAAAATGATATCAATTCAGGGAAGATGACGTTACTGCAAAGT 2520 TTCTTAAAGCAGGTAACGAACTGAAATTATATAAGACATTGTCATAAAGCAAAACAACAT 2580 GTACATGACATTGCCGCAAACATTGACTTATGCATAAGAAATACATCAGATGAGATAAGA 2640 AACTGAAAAAGTAATCTCATTTTTTCTACGGCAAATTTTACAAAGAAGACAATATGGGAA 2700 TAAAATAGACCATATTACCAAATTATGAAATGGACACTCTCTAAAGACTTATCAATCATT 2760 ACAAGCTGCTTTTAACGTGGAATTTTCTAAGTGTTCGACATTGATGTTGTTTTTATCACT 2820 ATCATCATATTTCCAAAGTTTTCAGGGTCGATGATTATAGAACAAGAGCTAACCTCAACG 2880 AGAGCACTGTTTGAAAGCACCCGACACCCCGAGGATTCAATAAAATTGAGGATAGAACGG 2940 TAAAATAGTAATCACTAACGAAGACATGCTAGAAATTTTTACATGAAAGACATTACATTT 3000 GCCCCTATGTTCAATGAACACGAATGAAATTTTGGATATACAACCACATTTTAAGTAATA 3060 GGTTATGGGTATAAAACTAAACTATAGCTAGTTGGTATTGGTAAAGGAGAAAGGTCCAAT 3120 GACATGATTTTGTCAATAAGTGCTTCAAGAAAATAATATTAGCCAACGGATAAGATTGAT 3180 CTAGACAGGAGCAATAACAACACGATAACACTGATAAATTGCAAGAACCAAGTAGGTCTC 3240 AATATTTGATGGTTATTATCACTCTTTCCCCAACAAAAGAATACAAATGTTCAAACCAAC 3300 ACCACTTCAGGGGAGGAGAATTTATGAATAAAATTGATATGAAGTAGTGGATGATTTGGA 3360 AAAAAATTCGGTAATGTTATGAATTTACCAATAGAATAAGATAACATTACTACCCCTGAT 3420 TGGTATATTAGACAGAACTTGTGATGATACTAAAACTCGAAAAGAGATGTTTCAAGCAGG 3480 GGCATATAATGTGCAAAATGTCACAATGACTCTAAGATTCAAATGGCATCATGATTTCTG 3540 AACTAAATACCAGCCAAACACTCTCGGTTGATACGTCCAATTTAAAAGACGGTCACCGAA 3600 AACAAAGTGAAAGTAAGTGAAATCATATGAATAGATTCACAAGAAATCGTAATGGAAACA 3660 CAATAATCATTTCATAAATCTTTGAAGTATACACAAGAATTGAGGTCAATAATTATCTAT 3720 TAAAAGACTACCACAAGGTTGACTTTATGATCCCAATACACCAATAGAGGAGATTGAAAC 3780 Follow-up ATCCTAGGTCAACGTCGAAGATGGAAATTTGAGACAAAGATGAATAAAAGGTCGACCGAT 3840 AACATTGTTTGATGAATCCAAAGAGAATTGGTACTTCAATACATTAAATAAAGATCTTCC 3900 GTTAAATGTAAGATACGAAATTGATGTGAATCCGTTACTTAAATATCTAAAGAAGAATAA 3960 TGCAACTTATGAGATATGTTGAAAACATGAATTAGCTTAAATCACACTAAAGTGAGAAGT 4020 TTGATTTTAAGAAACATACCATAGAGTATTTATAATATCGTCCATCCTACTGCAAAAAGA 4080 CTATTCCATGCTATACTCAAATCAAAAAACTGGGTACATACTTAATGATGATGACTAAAT 4140 TCAAACCAAGTGTCACCATTTCTAGATGGCTACTCAAAAATGAGAATGATGGCAAACAAA 200 TGGAAGATAGATCAATGTTTGAATTGCTTAGACAGAGATTTTGGCTAAAACACCAGCAGA 4260 CTATGAATGCGGAATACAAGTGATAAAGTTTTACAACTTCACCAAATTGTGAATATAATC 4320 AATCTGGAATCCCGGAAATAGAAATCAAAGATTGAAAATTTTTCGAAATCGACTAATAAA 4380 ATGCATGATTTCGTCAAATTTAAACAACACATACGCAAATATATTAAATTAGAATACCGA 4440 TGTGGATTGAAAATGGAGGTGAATACGTGTAAGTACGGTTAAAATTCCACAGTTTTAATT 4500 GTGTTTCATGGCATGTGTCAAGATTATCCATTTGCATAAACTTATCGAAATATTGGTAGT 4560 AAGGAAACAAGGTGAATGCCGGCCGGATTTACCAAGTACTGAAACTCATATTAGGTTTGG 4620 ATGTATGAAAATCGAGATATCTGATAACTACAGAAAACACATATACTACTTAAAAAATAC 4680 CCGAAGTCATTTTCATATTACAATCGGATAGTTAAACAAGGGGAGCTAACGCTCAAAGAG 4740 AGAGTCACATATTTGAATATGACTGTTGACACAGATCTTTATCTAGTGCGGGTTCATAGA 4800 TCTAGTGCTAAGAAAATAGATTGAGTTAATATGATCATTAGTGAATGAAGAAATGATAAC 4860 GTGAAGCGATGTTACAAGAAGATTACGATACAAAAGTGATACTTACGTTAAAATTATTCA 4920 GTATTTGCAAGCATATTAGATTAATAAGGTTAAAATTAACAAAAAACAATTAGGTCCATA 4980 AGCCCATATTACGAAATTATTCTTAGAAAAAGTTATGATTAATTTTCAATCCAACATTAC 5040 AATATTATCTCAAAATAAAGATGCTAGGTTAAAACACACTATTGTCCTTAGAGAAATGGA 5100 AGTTATGTGGAGTTAAAAAATAATCAGTGGAAAGAATCACCGACACAATGATTTGACATC 5160 Follow-up AATTGTTGATAACAAAGTCGCTAGAAGAGATTAAGGAATCAATGATGAACCATGATTACT 5220 GAATCAGGGAGTGTTCGCTATAGAGAGATTTCCTAGCCGGAATGCACGACAATCCTGAGA 5280 CGGAAGTCGATCGTCGATGCCCATGGTGCGTGGTGAAAAATTTTCTTAGAAAATTTGTTC 5340 TTTCCTTCAACTGCTTTTAAGAAAGAGAGGTTCAAGTGGTTTAAGTACGACGGTCACAAA 5400 GATTGCGGCTTATGAGGCCCGAACTGAGTTGAAATACAAAATCAAGATATAATTATATAC 5460 CTTACTTGTCCATATTGTTTTATAATACATTCTTCAGATATTTAAATTTCTGTGTATCAA 5520 CCTATAAAACAGAGATACATTCAGTGCATTTAGTATACTGAGTGAACTGGTACCTGTGAC 5580 ATTCAAGATAACTGTTTCGCGCACGCTGGCAGACGAACAATTGC 5624 ...

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