CN1427009A - Human zinc finger protein family new gene ZNF325 - Google Patents
Human zinc finger protein family new gene ZNF325 Download PDFInfo
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Abstract
A novel gene ZNF325 in human zinc finger protein family has 2697bp for the whole length of cDNA positioned at chromosome 7p22, 462 amino acids for the whole length of coded protein and 15 C2H2-type-contained zinc fingers. It has wide expression in adult and embryo stage, so playing important role in embryo development and normal function of human organs.
Description
Technical field:
The present invention relates to a kind of Human genome, specifically is the cDNA full-length clone of a human gene ZNF325 of zinc finger protein family, and other expression and expression in each organ of different developmental phases human embryos in each organ of adult.
Background technology:
Zinc finger protein family is a huge supergene family, and they are present in the various species widely, comprise yeast, fruit bat, Xenopus laevis, mouse and people.Zinc finger protein family generally has characteristic structure-----zinc finger protein structural domain of this family, be in fruit bat, be established at first, the necessary regulatory factors of tissue differentiation (Postigo et al.1997) such as muscle of unifying for central nervous system.Zinc finger protein gene zfh-1 expresses in the multiple mesoderm derived structure of the mesoderm of drosophila embryos and late embryo in early days, participates in regulating the formation of the heart precursor cell (EPC) of expressing the eve gene; Also together regulate the mesoblastic further differentiation in the outside that comprises the sexual gland mesoderm (Lai Z C., et al, 1999) with the tinman gene.The homologous gene of zfh-1 in vertebrates also has similar function.Expression product ZEB-1 and ZEB-2 albumen as zinc finger protein gene ZEB, in tissues such as muscle, central nervous system, T lymphocyte, expression is arranged all, and recent findings ZEB albumen comprises two distinct area, regulates Mammals myocyte and lymphocytic differentiation (Postigo et al.1997) respectively.
Zinc finger protein gene family also is one of human maximum gene family, and a lot of present known human transcription regulatory factors all are zinc finger protein member (Gou et al., 2001).Typical zinc finger protein is C2H2 type, i.e. Zn
++Ion and two and half Guang amino-acid residues and two histidine residues form coordinate bond.Except these four amino-acid residues, also has also high conservative of two die aromatischen Aminosaeuren residues and leucine residue.Coordination structure adds Phe and the formed hydrophobic core of Leu, has determined basic structure and shape that zinc refers to.The three-dimensional structure that zinc refers to comprises beta sheet and alpha-helix of two weak points.Studies show that zinc fingers is essential for the structure of keeping the dna structure territory, under many circumstances it also with dna direct interact (Coppin et al., 2002).It is believed that this zinc fingers works when protein bound is to Nucleotide, in the differentiation and reproductive process of cell, it can activate or the expression of suppressor gene, as zinc finger protein ZEB-1, snail, hFOG etc., and, this zinc fingers can be discerned identical dna sequence dna, as the zinc fingers of ZEB-1 and snail can discern they separately the E-BOX of target site and E-BOX similar sequence (Burke et al .2002).
Studies show that zinc finger protein gene serves as the role of housekeeping gene or regulatory factor in the biological development process, also have a lot of zinc finger proteins in human early embryonic development process, to play an important role.As ZNF174 is the negative regulon (Williams et al., 1995) of some growth factor gene; AJ18 participates in the growth (Jheon et al., 2001) of bone; Hsal1, WT1, and zinc finger protein such as TR-alpha-1 is relevant with heart function and disease, HFHZ may the pathology of cardiac hypertrophy be played an important role (Dai and Liew, 1998).Zinc finger protein GATA4 and hFOG2 participate in the formation of heart and the generation (Crispino et al., 2001) of disease.Many zinc finger proteins participate in the formation of some tumour, as WT1 (Dennis et al., 2002) and RB gene (Lee, et al., 1987).Simultaneously, in the animal pattern fruit bat, zinc finger protein Zfh1 participate in to regulate the eve expression of gene, and the formation of control heart precursor cell (EPC) is essential (Su et al., 1999) to the special differentiation of fruit bat heart.
Summary of the invention:
The object of the present invention is to provide the full length cDNA sequence of a new gene ZNF325 of human zinc finger protein family, analyze its expression in adult each organ and human embryos different developmental phases, with explore this gene in the human embryos growth course and adult grow in role.
The present invention is a template with the cDNA library of the brain mRNA reverse transcription gained that extract in this laboratory, conserved sequence region design degenerate primer A1 and A2 between referring to according to C2H2 type zinc fingers and adjacent zinc, carry out pcr amplification, the gained pcr amplification product separates through the swimming of sepharose point, the back conversion that is connected with pUCm-T (SANGONG) carrier enters bacillus coli DH 5 alpha, transforms a laggard pacing preface analysis.The sequence of gained is found it is the partial sequence of EST AU125576 with Blastn removal search dbEST, ESTAU125576 blastn EST removal search Genbank, we have found an EST group who comprises EST AU125576, they are by ESTs BG957653, BE568216, AV708178, AU125576 andAU131235 forms, and this EST group belongs to a new gene.With the embryo and brain library is template, uses four pairs of primers 5 '-RACE anti-sense-outer/UPM respectively, 5 '-RACEanti-sense-inner/UPM; 3 '-RACE sense-outer/NUP, 3 '-RACEsense-inner/NUP carry out 5 ｀-RACE and 3 ｀-RACE rapid amplifying end, 5 '-RACEanti-sense-outer wherein, 5 '-RACE anti-sense-inner; 3 '-RACE sense-outer, 3 '-RACE sense-inner is a gene specific primer, UPM and NUP are provided by SMART RACE test kit, and the RACE reaction has obtained the purpose fragment of 930bp and 318bp respectively, through T-carrier cloning, order-checking, they are the 5 ｀ end and the 3 ｀ end of this gene, they are spliced into full-length cDNA, are total to 2697bp, so far, we have obtained the full-length cDNA of new gene, through the approval called after ZNF325 of international man's genoid NK.
This gene of ZNF325 is positioned at karyomit(e) 7p22, and long 2697 bases contain 5 exons, long 15kb on genomic dna.Preceding 5 ' the end upstream of its open reading frame contains the non-coding region of 924bp, is open reading frame from 925bp to 2313bp, is 3 ' end non-coding sequence behind the 2313bp.462 amino acid of its encoded protein matter total length, the zinc that contains 15 C2H2 types refers to, it is a kind of typical Zinc finger domain, form by #-X-C-X (1-5)-C-X3-#-X5-#-X2-H-X (3-6)-[H/C], wherein X represents any amino acid, last amino acid is Histidine or halfcystine, and the # representative plays the amino acid position of important stable folding effect to zinc fingers.
The gene order of ZNF325 has been delivered to GenBank, and the number of accepting is AF505656.Analyze the same Feed Discovery of other zinc finger protein of itself and zinc finger protein family with the ALIGNMENT of JELLYFISH program, ZNF325 and RBAK homology degree are up to 71% (see figure 1).RBAK is first and interactional transcription inhibition factor of RB (Skkapek et al., 2000) that it is found that.And RB is a cancer suppressor gene that can suppress the genetic expression of E2F transcription factor family member activated.
This gene of Northern Bolt analysis revealed is generally expressed in each tissue of adult.Carry out the Northern hybridization analysis of 8 kinds of normal human tissue mRNA, the result generally expresses (seeing Fig. 2 A) for the ZNF325 mRNA of 2.7kb in each tissue of adult, this result shows that the expression of ZNF325 in adult do not have cell-specific, and it may all play an important role to the normal function of each tissue of human body.
Carried out for 16 weeks, 18 weeks, 20.5 week, 22 weeks, the Northern hybridization analysis of the various total tissue RNA films of human embryo in 23 weeks and 24 weeks, the result shows that this gene is similar to the expression at adult at the embryo's of human body different development stage expression, and ZNF325 mRNA illustrates that at embryo's wide expression in period (seeing Fig. 2 B-2G) ZNF325 may play vital role to the normal development of human body early embryo.What its structure and expression characteristic were indicating its coding is the transcriptional regulator of a tool DNA combined function.
Advantage of the present invention:
1. the full length cDNA sequence of a human gene ZNF325 gene is provided, and has described the homology of its encoded protein product and important gene RBAK, proved that this gene is C
2H
2One of member of type zinc finger protein gene family.
2. the protein product of this genes encoding is a zinc finger protein, it is a kind of transcriptional regulator, and wide expression in human adult and embryonic tissue, it is very important to illustrate that this gene has play a part in fetal development in early days as transcription factor, and this effect has been extended to adult period.
The present invention is further elaborated by the following drawings and embodiment, but does not limit the scope of the invention.
Description of drawings:
The proteinic homology of Fig. 1 .ZNF325 and RBAK relatively.
Fig. 2 .ZNF325 is in the Northern of human tissue hybridization.
Fig. 2 A, 2B, 2C, 2D, 2E, 2F, 2G are respectively adult's tissue, 16 weeks, 18 weeks, 20.5 weeks, 22 weeks, 23 weeks, 24 all embryonic tissues.
" cerebellum 6 intestines 7 livers 8 hearts 9 suprarenal gland 10 pancreas 11 spleens 12 lungs 13 gall-bladders 14 bladders 15 tongues among the figure: 1 stomach, 2 kidneys, 3 skeletal muscle, 4 lungs, 5 brains, 5 ' brain 5
Embodiment: the clone of the new gene ZNF325 of human zinc finger protein family, order-checking and expression thereof.
1. degenerate primer carries out pcr amplification and screens new gene
Conserved sequence region design degenerate primer A1 and A2A1:sense 5 '-GRNGARAARCCNUMY-3 ' A2:antisense 5 '-RTGDATNCGNTGRTGVAW-3 ' PCR reaction between referring to according to C2H2 type zinc fingers and adjacent zinc are template with brain cDNA library, reaction conditions: 94 ℃ of sex change of elder generation 4 minutes, 30 seconds, 50 ℃ 30 seconds, 72 ℃ totally 30 circulations in 30 seconds of 94 ℃ of sex change then, last 72 ℃ were extended 7 minutes, the band of amplification, carrying out the T-carrier connects, transform, select transformant arbitrarily and go order-checking.
Sequencing result goes to carry out the dbEST retrieval; Finds that this fragment is the part fragment of EST AU125576; Carries out Blastn nr retrieval with this EST and obtains a new gene, the cDNA sequences Design RACE primer that the clone of carries out the dbEST retrieval again.2.ZNF325 full-length cDNA obtains with retrieval: 5 '-RACE anti-sense-outer, 5 '-CTGATGCACAGTAAGGGTTCC-3 ' 5 '-RACE anti-sense-inner 5 '-AGGTCCGACATCTGGAGAAAGGC-3 ' 3 '-RACE sense-outer 5 '-TGATGTATAGTGAGATAGGACC-3 ' 3 '-RACE sense-inner 5 '-TCAGAGAAGCCTTCCCACATC-3 ' utilizes the SMART RACE kit of CLONTECH company to do the primer that 5 ' end and 3 ' end CLONTECH company provides: MUP:5 '-CTAATACGACTCACTATAGGGCAAGCAGTGGTATCAAC
GCAGAGT-3′
5′-CTAATACGACTCACTATAGGGC-3′NUP：5′-AAGCAGTGGTATCAACGCAGAGT-3′
Concrete operations are undertaken by the protocol of the said firm.The RACE product cloning is gone into 5 ｀ end and the 3 ｀ terminal sequences of T carrier and order-checking acquisition ZNF325, reaction has obtained the purpose fragment of 930bp and 318bp respectively, will obtain the ZNF325 full-length cDNA after their splicings, altogether 2697bp, so far, we have obtained the full-length cDNA of new gene.3.Northern the extraction of hybridization (1) total tissue RNA (press Wang Yanyun etc., a kind of method for extracting total RNA fast, life science, 5 (1): 88-90 (2001) carries out).(2) electrophoresis of total RNA and commentaries on classics film, all the method for introducing by " modern molecular biology experimental technique " is carried out (Lu Shengdong chief editor, second edition).(3) Northern hybridization
Will be from CLONTECH company the adult's tissue mRNA film bought and the nylon membrane that changes total RNA that the multiple tissue of 20ug human embryo and total embryonic tissue are arranged, put into culture dish, add sex change through the quick hybridization liquid 10ml of 42 ℃ of preheatings (Clontech company), prehybridization is 30 minutes under 42 ℃ of conditions, add the cDNA fragment of the isotope-labeled ZNF325 of sex change, 42 ℃ of hybridization are spent the night.With 2 * SSC, 0.05%SDS, washed twice, 15 minute/time for 42 ℃; 0.1 * SSC, 0.1%SDS washed twice, 15 minute/time for 42 ℃.(4)-80 ℃ exposure is 3 days, X-ray sheet developing.(5) wash 10 minutes films with β-actin cDNA mark again with 0.1 * SSC and 0.5%SDS at 95 ℃, do Northern hybridization carrying out positive control.
ZNF325 cDNA：accaggtctg gtctacagtc gactaagtcg gtcggcccgg gcccctctgg agcanaangg 60ccttggtgag gcggacagga agggacctcg cgagcagacg cgcgcccgcg acagcaatcc 120cgccccggcc tgtcgggagc ggtggggcag aggctgcgga gccccaggag gatctgcctc 180cgctttcaca gtcctccaga tttttccaag agcagcagaa aatgaataaa tccctggggc 240cagtgtcatt caaggacgtg gctgtggact tcacccagga ggaatggcag cagctggatc 300ctgagcagaa gataacttac agggatgtga tgctggagaa ctacagcaat ctagtttctg 360tggggtatca cattatcaaa ccggatgtta tcagcaagtt ggagcaagga gaagagccat 420ggatagtaga aggagaattc ctacttcaga gctatccaga tgaagtctgg caaactgatg 480acctaataga gagaatccag gaagaggaaa ataaaccttc aaggcaaact gtgttcattg 540agaccctgat tgaagagaga ggtaatgttc ctggtaaaac ttttgatgta gaaacgaacc 600ctgttccttc aagaaaaata gcctataaaa atagcctctg tgactcatgt gaaaagtgtt 660taacgtctgt ttcagaatat attagtagtg atggaagcta tgcaagaatg aaagctgatg 720aatgtagtgg attgtgggaa atcactcctc catattaagc ttgagaaaac tcatccagga 780gatcnagctt atgatttaat caaaatgggg aaccttatac cctaaattga anaagtntta 840tcaaaaattc gtattttgga gaaacctttt gaatatattg aatgccagaa agccttccaa 900aaggacactg tttttgttaa tcac atg gaa gaa aag ccc tat aag 945
Met?Glu?Glu?Lys?Pro?Tyr?Lys
1 5 tgg?aat?gga?tct?gaa?ata?gcc?ttt?ctc?cag?atg?tcg?gac?ctc?act 990 Trp?Asn?Gly?Ser?Glu?Ile?Ala?Phe?Leu?Gln?Met?Ser?Asp?Leu?Thr
10 15 20 gta?cat?cag?aca?tct?cat?atg?gaa?atg?aag?ccc?tat?gaa?tgc?agt 1035 Val?His?Gln?Thr?Ser?His?Met?Glu?Met?Lys?Pro?Tyr?Glu?Cys?Ser
25 30 35 gaa?tgt?ggg?aaa?tcc?ttc?tgt?aaa?aag?tca?aaa?ttt?att?ata?cat 1080 Glu?Cys?Gly?Lys?Ser?Phe?Cys?Lys?Lys?Ser?Lys?Phe?Ile?Ile?His
40 45 50 cag?agg?act?cac?aca?gga?gag?aaa?cct?tac?gaa?tgt?aat?cag?tgt 1125 Gln?Arg?Thr?His?Thr?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Asn?Gln?Cys
55 60 65 ggg?aaa?tcc?ttc?tgc?cag?aag?gga?acc?ctt?act?gtg?cat?cag?aga 1170 Gly?Lys?Ser?Phe?Cys?Gln?Lys?Gly?Thr?Leu?Thr?Val?His?Gln?Arg
70 75 80 aca?cac?aca?ggg?gag?aag?ccc?tat?gaa?tgt?aat?gaa?tgt?ggg?aag 1215 Thr?His?Thr?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Asn?Glu?Cys?Gly?Lys
85 90 95aac?ttt?tac?cag?aag?tta?cac?ctc?att?cag?cat?cag?aga?act?cac 1260Asn?Phe?Tyr?Gln?Lys?Leu?His?Leu?Ile?Gln?His?Gln?Arg?Thr?His
100 105 110tca?gga?gag?aag?ccc?tat?gaa?tgt?agt?tat?tgt?gga?aaa?tcc?ttt 1305Ser?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Ser?Tyr?Cys?Gly?Lys?Ser?Phe
115 120 125tgc?cag?aag?aca?cac?ctc?aca?caa?cat?cag?aga?aca?cat?tca?gga 1350Cys?Gln?Lys?Thr?His?Leu?Thr?Gln?His?Gln?Arg?Thr?His?Ser?Gly
130 135 140gag?aga?cct?tat?gtt?tgt?cat?gac?tgt?ggg?aaa?acc?ttc?tcg?cag 1395Glu?Arg?Pro?Tyr?Val?Cys?His?Asp?Cys?Gly?Lys?Thr?Phe?Ser?Gln
145 150 155aag?tca?gca?ctt?aat?gac?cat?cag?aaa?att?cac?aca?ggt?gtg?aaa 1440Lys?Ser?Ala?Leu?Asn?Asp?His?Gln?Lys?Ile?His?Thr?Gly?Val?Lys
160 165 170ctc?tac?aag?tgt?agt?gaa?tgt?ggg?aaa?tgc?ttc?tgc?cgc?aag?tct 1485Leu?Tyr?Lys?Cys?Ser?Glu?Cys?Gly?Lys?Cys?Phe?Cys?Arg?Lys?Ser
175 180 185act?ctc?acg?acc?cac?ctg?agg?acc?cac?aca?gga?gag?aaa?ccg?tat 1530Thr?Leu?Thr?Thr?His?Leu?Arg?Thr?His?Thr?Gly?Glu?Lys?Pro?Tyr
190 195 200gaa?tgt?aat?gag?tgt?gga?aaa?ttc?ttc?tct?cgg?ttg?tca?tat?ctc 1575Glu?Cys?Asn?Glu?Cys?Gly?Lys?Phe?Phe?Ser?Arg?Leu?Ser?Tyr?Leu
205 210 215act?gta?cat?tat?aga?act?cat?tca?gga?gag?aaa?ccc?tat?gaa?tgt 1620Thr?Val?His?Tyr?Arg?Thr?His?Ser?Gly?Glu?Lys?Pro?Tyr?Glu?Cys
220 225 230aat?gaa?tgt?gga?aaa?acc?ttc?tac?ctg?aat?tca?gcc?ctc?atg?aga 1665Asn?Glu?Cys?Gly?Lys?Thr?Phe?Tyr?Leu?Asn?Ser?Ala?Leu?Met?Arg
235 240 245cat?cag?aga?gtg?cac?aca?gga?gag?aaa?cct?tac?gaa?tgt?aat?gaa 1710His?Gln?Arg?Val?His?Thr?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Asn?Glu
250 255 260tgt?gga?aag?tta?ttc?tcc?cag?ttg?tca?tac?ctc?act?atc?cat?cat 1755Cys?Gly?Lys?Leu?Phe?Ser?Gln?Leu?Ser?Tyr?Leu?Thr?Ile?His?His
265 270 275aga?act?cat?tca?gga?gta?aaa?ccc?tat?gaa?tgt?agt?gaa?tgt?ggg 1800Arg?Thr?Lis?Ser?Gly?Val?Lys?Pro?Tyr?Glu?Cys?Ser?Glu?Cys?Gly
280 285 290aaa?acc?ttc?tac?cag?aac?tca?gcc?ctt?tgt?aga?cat?cgg?aga?ata 1845Lys?Thr?Phe?Tyr?Gln?Asn?Ser?Ala?Leu?Cys?Arg?His?Arg?Arg?Ile
295 300 305cac?aaa?gga?gag?aag?ccc?tat?gaa?tgc?tat?ata?tgt?gga?aaa?ttc 1890His?Lys?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Tyr?Ile?Cys?Gly?Lys?Phe
310 315 320ttc?tct?cag?atg?tca?tac?ctc?act?ata?cat?cat?aga?att?cat?tca 1935Phe?Ser?Gln?Met?Ser?Tyr?Leu?Thr?Ile?His?His?Arg?Ile?His?Ser
325 330 335gga?gag?aag?ccc?tat?gaa?tgt?agt?gaa?tgt?ggg?aaa?acc?ttc?tgc 1980Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Ser?Glu?Cys?Gly?Lys?Thr?Phe?Cys
340 345 350cag?aat?tca?gcc?ctt?aat?cga?cat?cag?aga?aca?cac?aca?gga?gag 2025Gln?Asn?Ser?Ala?Leu?Asn?Arg?His?Gln?Arg?Thr?His?Thr?Gly?Glu
355 360 365aaa?gcc?tac?gaa?tgt?tat?gaa?tgt?ggg?aag?tgc?ttc?tct?cag?atg 2070Lys?Ala?Tyr?Glu?Cys?Tyr?Glu?Cys?Gly?Lys?Cys?Phe?Ser?Gln?Met
370 375 380tcc?tat?ctc?act?ata?cat?cat?cga?att?cat?tca?gga?gag?aaa?ccc 2115Ser?Tyr?Leu?Thr?Ile?His?His?Arg?Ile?His?Ser?Gly?Glu?Lys?Pro
385 390 395ttt?gaa?tgt?aat?gag?tgt?gga?aaa?gcc?ttc?tct?cgg?atg?tca?tac 2160Phe?Glu?Cys?Asn?Glu?Cys?Gly?Lys?Ala?Phe?Ser?Arg?Met?Ser?Tyr
400 405 410ctc?act?gta?cat?tat?aga?act?cat?tca?gga?gag?aaa?ccc?tat?gag 2205Leu?Thr?Val?His?Tyr?Arg?Thr?His?Ser?Gly?Glu?Lys?Pro?Tyr?Glu
415 420 425tgt?act?gaa?tgt?gga?aaa?aaa?ttc?tac?cac?aaa?tca?gca?ttc?aac 2250Cys?Thr?Glu?Cys?Gly?Lys?Lys?Phe?Tyr?His?Lys?Ser?Ala?Phe?Asn
430 435 440agc?cat?cag?aga?att?cat?agg?aga?ggc?aat?atg?aat?gta?ata?gat 2295Ser?His?Gln?Arg?Ile?His?Arg?Arg?Gly?Asn?Met?Asn?Val?Ile?Asp
445 450 455gtg?gga?agg?ctt?ctc tgaagtcaga?cctcatttta?tatcagagaa 2340Val?Gly?Arg?Leu?Leu
460 462ccctttcagt atagtgaatc agaaactcct gcctgaagtc aaacaccttg tacatcagag 2400agttcacaca ggttagtgtg gacatcccct tgtgtgttgg actcataatc tgaagactca 2461cagaatggaa accatgatta taacaagacc acatggtata acaatactag actatagaca 2520agtaaaaatt tataaatatt aagaatgtat atacatgtca ccatggattg gaactgtttt 2580gcatatcagg gaaatcatag ccaaggggaa atctatcagt ataaggaatg tggaagacat 2640aatcctttgg aaactgttaa tactaaaaga tatgtttctg atacaatagc aaacttg 2697ZNF325 cDNAaccaggtctg gtctacagtc gactaagtcg gtcggcccgg gcccctctgg agcanaangg 60ccttggtgag gcggacagga agggacctcg cgagcagacg cgcgcccgcg acagcaatcc 120cgccccggcc tgtcgggagc ggtggggcag aggctgcgga gccccaggag gatctgcctc 180cgctttcaca gtcctccaga tttttccaag agcagcagaa aatgaataaa tccctggggc 240cagtgtcatt caaggacgtg gctgtggact tcacccagga ggaatggcag cagctggatc 300ctgagcagaa gataacttac agggatgtga tgctggagaa ctacagcaat ctagtttctg 360tggggtatca cattatcaaa ccggatgtta tcagcaagtt ggagcaagga gaagagccat 420ggatagtaga aggagaattc ctacttcaga gctatccaga tgaagtctgg caaactgatg 480acctaataga gagaatccag gaagaggaaa ataaaccttc aaggcaaact gtgttcattg 540agaccctgat tgaagagaga ggtaatgttc ctggtaaaac ttttgatgta gaaacgaacc 600ctgttccttc aagaaaaata gcctataaaa atagcctctg tgactcatgt gaaaagtgtt 660taacgtctgt ttcagaatat attagtagtg atggaagcta tgcaagaatg aaagctgatg 720aatgtagtgg attgtgggaa atcactcctc catattaagc ttgagaaaac tcatccagga 780gatcnagctt atgatttaat caaaatgggg aaccttatac cctaaattga anaagtntta 840tcaaaaattc gtattttgga gaaacctttt gaatatattg aatgccagaa agccttccaa 900aaggacactg tttttgttaa tcacatggaa gaaaagccct ataagtggaa tggatctgaa 960atagcctttc tccagatgtc ggacctcact gtacatcaga catctcatat ggaaatgaag 1020ccctatgaat gcagtgaatg tgggaaatcc ttctgtaaaa agtcaaaatt tattatacat 1080cagaggactc acacaggaga gaaaccttac gaatgtaatc agtgtgggaa atccttctgc 1140cagaagggaa cccttactgt gcatcagaga acacacacag gggagaagcc ctatgaatgt 1200aatgaatgtg ggaagaactt ttaccagaag ttacacctca ttcagcatca gagaactcac 1260tcaggagaga agccctatga atgtagttat tgtggaaaat ccttttgcca gaagacacac 1320ctcacacaac atcagagaac acattcagga gagagacctt atgtttgtca tgactgtggg 1380aaaaccttct cgcagaagtc agcacttaat gaccatcaga aaattcacac aggtgtgaaa 1440ctctacaagt gtagtgaatg tgggaaatgc ttctgccgca agtctactct cacgacccac 1500ctgaggaccc acacaggaga gaaaccgtat gaatgtaatg agtgtggaaa attcttctct 1560cggttgtcat atctcactgt acattataga actcattcag gagagaaacc ctatgaatgt 1620aatgaatgtg gaaaaacctt ctacctgaat tcagccctca tgagacatca gagagtgcac 1680acaggagaga aaccttacga atgtaatgaa tgtggaaagt tattctccca gttgtcatac 1740ctcactatcc atcatagaac tcattcagga gtaaaaccct atgaatgtag tgaatgtggg 1800aaaaccttct accagaactc agccctttgt agacatcgga gaatacacaa aggagagaag 1860ccctatgaat gctatatatg tggaaaattc ttctctcaga tgtcatacct cactatacat 1920catagaattc attcaggaga gaagccctat gaatgtagtg aatgtgggaa aaccttctgc 1980cagaattcag cccttaatcg acatcagaga acacacacag gagagaaagc ctacgaatgt 2040tatgaatgtg ggaagtgctt ctctcagatg tcctatctca ctatacatca tcgaattcat 2100tcaggagaga aaccctttga atgtaatgag tgtggaaaag ccttctctcg gatgtcatac 2160ctcactgtac attatagaac tcattcagga gagaaaccct atgagtgtac tgaatgtgga 2220aaaaaattct accacaaatc agcattcaac agccatcaga gaattcatag gagaggcaat 2280atgaatgtaa tagatgtggg aaggcttctc tgaagtcaga cctcatttta tatcagagaa 2340ccctttcagt atagtgaatc agaaactcct gcctgaagtc aaacaccttg tacatcagag 2400agttcacaca ggttagtgtg gacatcccct tgtgtgttgg actcataatc tgaagactca 2460cagaatggaa accatgatta taacaagacc acatggtata acaatactag actatagaca 2520agtaaaaatt tataaatatt aagaatgtat atacatgtca ccatggattg gaactgtttt 2580gcatatcagg gaaatcatag ccaaggggaa atctatcagt ataaggaatg tggaagacat 2640aatcctttgg aaactgttaa tactaaaaga tatgtttctg atacaatagc aaacttg 2697
Claims (1)
1、ZNF325,cDNA：accaggtctg gtctacagtc gactaagtcg gtcggcccgg gcccctctgg agcanaangg 60ccttggtgag gcggacagga agggacctcg cgagcagacg cgcgcccgcg acagcaatcc 120cgccccggcc tgtcgggagc ggtggggcag aggctgcgga gccccaggag gatctgcctc 180cgctttcaca gtcctccaga tttttccaag agcagcagaa aatgaataaa tccctggggc 240cagtgtcatt caaggacgtg gctgtggact tcacccagga ggaatggcag cagctggatc 300ctgagcagaa gataacttac agggatgtga tgctggagaa ctacagcaat ctagtttctg 360tggggtatca cattatcaaa ccggatgtta tcagcaagtt ggagcaagga gaagagccat 420ggatagtaga aggagaattc ctacttcaga gctatccaga tgaagtctgg caaactgatg 480acctaataga gagaatccag gaagaggaaa ataaaccttc aaggcaaact gtgttcattg 540agaccctgat tgaagagaga ggtaatgttc ctggtaaaac ttttgatgta gaaacgaacc 600ctgttccttc aagaaaaata gcctataaaa atagcctctg tgactcatgt gaaaagtgtt 660taacgtctgt ttcagaatat attagtagtg atggaagcta tgcaagaatg aaagctgatg 720aatgtagtgg attgtgggaa atcactcctc catattaagc ttgagaaaac tcatccagga 780gatcnagctt atgatttaat caaaatgggg aaccttatac cctaaattga anaagtntta 840tcaaaaattc gtattttgga gaaacctttt gaatatattg aatgccagaa agccttccaa 900aaggacactg tttttgttaa tcac atg gaa gaa aag ccc tat aag 945
Met?Glu?Glu?Lys?Pro?Tyr?Lys
1 5?tgg?aat?gga?tct?gaa?ata?gcc?ttt?ctc?cag?atg?tcg?gac?ctc?act 990?Trp?Asn?Gly?Ser?Glu?Ile?Ala?Phe?Leu?Gln?Met?Ser?Asp?Leu?Thr
10 15 20?gta?cat?cag?aca?tct?cat?atg?gaa?atg?aag?ccc?tat?gaa?tgc?agt 1035?Val?His?Gln?Thr?Ser?His?Met?Glu?Met?Lys?Pro?Tyr?Glu?Cys?Ser
25 30 35?gaa?tgt?ggg?aaa?tcc?ttc?tgt?aaa?aag?tca?aaa?ttt?att?ata?cat 1080?Glu?Cys?Gly?Lys?Ser?Phe?Cys?Lys?Lys?Ser?Lys?Phe?Ile?Ile?His
40 45 50cag?agg?act?cac?aca?gga?gag?aaa?cct?tac?gaa?tgt?aat?cag?tgt 1125Gln?Arg?Thr?His?Thr?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Asn?Gln?Cys
55 60 65ggg?aaa?tcc?ttc?tgc?cag?aag?gga?acc?ctt?act?gtg?cat?cag?aga 1170Gly?Lys?Ser?Phe?Cys?Gln?Lys?Gly?Thr?Leu?Thr?Val?His?Gln?Arg
70 75 80aca?cac?aca?ggg?gag?aag?ccc?tat?gaa?tgt?aat?gaa?tgt?ggg?aag 1215Thr?His?Thr?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Asn?Glu?Cys?Gly?Lys
85 90 95aac?ttt?tac?cag?aag?tta?cac?ctc?att?cag?cat?cag?aga?act?cac 1260Asn?Phe?Tyr?Gln?Lys?Leu?His?Leu?Ile?Gln?His?Gln?Arg?Thr?His
100 105 110tca?gga?gag?aag?ccc?tat?gaa?tgt?agt?tat?tgt?gga?aaa?tcc?ttt 1305Ser?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Ser?Tyr?Cys?Gly?Lys?Ser?Phe
115 120 125tgc?cag?aag?aca?cac?ctc?aca?caa?cat?cag?aga?aca?cat?tca?gga 1350Cys?Gln?Lys?Thr?His?Leu?Thr?Gln?His?Gln?Arg?Thr?His?Ser?Gly
130 135 140gag?aga?cct?tat?gtt?tgt?cat?gac?tgt?ggg?aaa?acc?ttc?tcg?cag 1395Glu?Arg?Pro?Tyr?Val?Cys?His?Asp?Cys?Gly?Lys?Thr?Phe?Ser?Gln
145 150 155aag?tca?gca?ctt?aat?gac?cat?cag?aaa?att?cac?aca?ggt?gtg?aaa 1440Lys?Ser?Ala?Leu?Asn?Asp?His?Gln?Lys?Ile?His?Thr?Gly?Val?Lys
160 165 170ctc?tac?aag?tgt?agt?gaa?tgt?ggg?aaa?tgc?ttc?tgc?cgc?aag?tct 1485Leu?Tyr?Lys?Cys?Ser?Glu?Cys?Gly?Lys?Cys?Phe?Cys?Arg?Lys?Ser
175 180 185act?ctc?acg?acc?cac?ctg?agg?acc?cac?aca?gga?gag?aaa?ccg?tat 1530Thr?Leu?Thr?Thr?His?Leu?Arg?Thr?His?Thr?Gly?Glu?Lys?Pro?Tyr
190 195 200gaa?tgt?aat?gag?tgt?gga?aaa?ttc?ttc?tct?cgg?ttg?tca?tat?ctc 1575Glu?Cys?Asn?Glu?Cys?Gly?Lys?Phe?Phe?Ser?Arg?Leu?Ser?Tyr?Leu
205 210 215act?gta?cat?tat?aga?act?cat?tca?gga?gag?aaa?ccc?tat?gaa?tgt 1620Thr?Val?His?Tyr?Arg?Thr?His?Ser?Gly?Glu?Lys?Pro?Tyr?Glu?Cys
220 225 230aat?gaa?tgt?gga?aaa?acc?ttc?tac?ctg?aat?tca?gcc?ctc?atg?aga 1665Asn?Glu?Cys?Gly?Lys?Thr?Phe?Tyr?Leu?Asn?Ser?Ala?Leu?Met?Arg
235 240 245cat?cag?aga?gtg?cac?aca?gga?gag?aaa?cct?tac?gaa?tgt?aat?gaa 1710His?Gln?Arg?Val?His?Thr?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Asn?Glu
250 255 260tgt?gga?aag?tta?ttc?tcc?cag?ttg?tca?tac?ctc?act?atc?cat?cat 1755Cys?Gly?Lys?Leu?Phe?Ser?Gln?Leu?Ser?Tyr?Leu?Thr?Ile?His?His
265 270 275aga?act?cat?tca?gga?gta?aaa?ccc?tat?gaa?tgt?agt?gaa?tgt?ggg 1800Arg?Thr?His?Ser?Gly?Val?Lys?Pro?Tyr?Glu?Cys?Ser?Glu?Cys?Gly
280 285 290aaa?acc?ttc?tac?cag?aac?tca?gcc?ctt?tgt?aga?cat?cgg?aga?ata 1845Lys?Thr?Phe?Tyr?Gln?Asn?Ser?Ala?Leu?Cys?Arg?His?Arg?Arg?Ile
295 300 305cac?aaa?gga?gag?aag?ccc?tat?gaa?tgc?tat?ata?tgt?gga?aaa?ttc 1890His?Lys?Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Tyr?Ile?Cys?Gly?Lys?Phe
310 315 320ttc?tct?cag?atg?tca?tac?ctc?act?ata?cat?cat?aga?att?cat?tca 1935Phe?Ser?Gln?Met?Ser?Tyr?Leu?Thr?Ile?His?His?Arg?Ile?His?Ser
325 330 335gga?gag?aag?ccc?tat?gaa?tgt?agt?gaa?tgt?ggg?aaa?acc?ttc?tgc 1980Gly?Glu?Lys?Pro?Tyr?Glu?Cys?Ser?Glu?Cys?Gly?Lys?Thr?Phe?Cys
340 345 350cag?aat?tca?gcc?ctt?aat?cga?cat?cag?aga?aca?cac?aca?gga?gag 2025Gln?Asn?Ser?Ala?Leu?Asn?Arg?His?Gln?Arg?Thr?His?Thr?Gly?Glu
355 360 365aaa?gcc?tac?gaa?tgt?tat?gaa?tgt?ggg?aag?tgc?ttc?tct?cag?atg 2070Lys?Ala?Tyr?Glu?Cys?Tyr?Glu?Cys?Gly?Lys?Cys?Phe?Ser?Gln?Met
370 375 380tcc?tat?ctc?act?ata?cat?cat?cga?att?cat?tca?gga?gag?aaa?ccc 2115Ser?Tyr?Leu?Thr?Ile?His?His?Arg?Ile?His?Ser?Gly?Glu?Lys?Pro
385 390 395ttt?gaa?tgt?aat?gag?tgt?gga?aaa?gcc?ttc?tct?cgg?atg?tca?tac 2160Phe?Glu?Cys?Asn?Glu?Cys?Gly?Lys?Ala?Phe?Ser?Arg?Met?Ser?Tyr
400 405 410ctc?act?gta?cat?tat?aga?act?cat?tca?gga?gag?aaa?ccc?tat?gag 2205Leu?Thr?Val?His?Tyr?Arg?Thr?His?Ser?Gly?Glu?Lys?Pro?Tyr?Glu
415 420 425tgt?act?gaa?tgt?gga?aaa?aaa?ttc?tac?cac?aaa?tca?gca?ttc?aac 2250Cys?Thr?Glu?Cys?Gly?Lys?Lys?Phe?Tyr?His?Lys?Ser?Ala?Phe?Asn
430 435 440agc?cat?cag?aga?att?cat?agg?aga?ggc?aat?atg?aat?gta?ata?gat 2295Ser?His?Gln?Arg?Ile?His?Arg?Arg?Gly?Asn?Met?Asn?Val?Ile?Asp
445 450 455gtg?gga?agg?ctt?ctc tgaagtcaga?cctcatttta?tatcagagaa 2340Val?Gly?Arg?Leu?Leu
460 462ccctttcagt?atagtgaatc?agaaactcct?gcctgaagtc?aaacaccttg?tacatcagag?2400agttcacaca?ggttagtgtg?gacatcccct?tgtgtgttgg?actcataatc?tgaagactca?2461cagaatggaa?accatgatta?taacaagacc?acatggtata?acaatactag?actatagaca?2520agtaaaaatt?tataaatatt?aagaatgtat?atacatgtca?ccatggattg?gaactgtttt?2580gcatatcagg?gaaatcatag?ccaaggggaa?atctatcagt?ataaggaatg?tggaagacat?2640aatcctttgg?aaactgttaa?tactaaaaga?tatgtttctg?atacaatagc?aaacttg 2697
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