CN101712958A - Novel human tumor inhibiting gene KIAA0157 and application thereof - Google Patents
Novel human tumor inhibiting gene KIAA0157 and application thereof Download PDFInfo
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- CN101712958A CN101712958A CN200810167194A CN200810167194A CN101712958A CN 101712958 A CN101712958 A CN 101712958A CN 200810167194 A CN200810167194 A CN 200810167194A CN 200810167194 A CN200810167194 A CN 200810167194A CN 101712958 A CN101712958 A CN 101712958A
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Abstract
The invention provides a novel human tumor inhibiting gene and an encoded protein thereof. The gene and the encoded protein can inhibit the growth of tumor cells, are down-regulated in various tumor tissues, and have a close relation with the generation of tumors. The gene and the encoded protein and derivatives thereof can be used for the diagnosis, treatment and drug screening of various tumors.
Description
Technical field
The present invention relates to a kind of new human tumor suppressor gene, also relate to the protein and the application in diagnosing tumor and treatment of the DNA that forms this gene and this dna encoding.
Background of invention:
Tumour is that environmental carcinogenesis factor and host gene interact, and causes gradually and constantly accumulates the unusual of series of genes, causes the result of cell proliferation and dead running balance imbalance.Develop into life-threatening aggressive malignant tumour from normal cell, need through the precancerous lesion stage mostly.Malignant tumour be the process that progressively develops a multistage, be broadly divided into excite, promote, several stages such as progress and transfer.In canceration multistage property evolution process, the sudden change that has often accumulated series of genes can relate to the variation of several genes on the coloured differently body, comprising: oncogene, cancer suppressor gene, injury repairing genes involved, cell cycle control gene etc.Studies show that, the generation of tumour, relate in the evolution because the activation of the oncogene that genetically deficient or sudden change cause and the deactivation of tumor suppressor gene, wherein the inactivation of cancer suppressor gene is a recurrent incident in most of tumour cells. at present, to comprising RB1, P53, the understanding of a plurality of cancer suppressor genes of WT1 etc. is very deep, these researchs have not only disclosed the molecular mechanism of tumor development, also provide target spot for Zao Qizhenduan Fen Zifenxing medicine research and development of tumour etc. therefore, the research of this respect is one of focus of academia always, and new cancer suppressor gene also just constantly obtains finding and using.
As hereinafter describing, in the research of THAP11 protein interactive protein matter, the present inventor find some gene coded product can with the THAP11 protein interaction, and system has identified this gene after separation.KIAA0157 is positioned 10q26.13 (NM_032182.), mRNA total length 3008bp, and cDNA1248bp, 416 amino acid (sequence 2) of encoding, molecular weight is the protein of 47KD.Mouse, rat and people's KIAA0157 homology illustrates its high conservative during evolution up to more than 90%, may bring into play important biological function.Utilize bioinformatic analysis to show, KIAA0157 active site of protein 215-266 amino acids place exists a coiled coil structural domain, this structural domain is present in the monomeric protein usually, and high conservative, mediates the interaction between albumen and the albumen usually.In the interaction protein screening process to inhibitor of apoptosis protein BPE (BRCA complex subunit 4), KIAA0157 is screened to be arrived.But, do not see any research report so far as yet at present to its function.
Summary of the invention
The present invention is cloned into the KIAA0157 full-length cDNA from people's marrow library, and utilizes the GFP-KIAA0157 fusion protein technology to detect the cellular localization of KIAA0157, and the result shows that KIAA0157 is full cell distribution (Fig. 2).Utilize the cDNA micro-array chip to detect and show that KIAA0157 is expressed in multiple tissue, and obviously downward modulation expression (Fig. 3) in the kinds of tumors tissue; In addition, utilize Real-time PCR to detect the expression of KIAA0157 in the 12 routine hepatocarcinoma patients, show that KIAA0157 expression level detail in liver cancer tissue is higher than cancer beside organism (Fig. 4) in 10 routine patients.High expression level KIAA0157 can suppress to comprise liver cancer cell lung carcinoma cell the growth (Fig. 5) of neuroblastoma cell, and cause cell that the G0/G1 phase takes place blocking (Fig. 6).Therefore monopolist of the present invention reaches a conclusion, and this gene may be a kind of new tumor suppressor gene.
Therefore, the invention provides a kind of DNA that constitutes people's gene, have sequence 1 described nucleotide sequence or the nucleotide sequence of one or more base deletions, replacement or increase is arranged.
The invention provides a kind of protein, the aminoacid sequence that it has aminoacid sequence shown in the sequence 2 or one or more aminoacid deletion is arranged, substitute or increase with respect to described aminoacid sequence.
The invention provides a kind of probe, it comprises the DNA with the DNA hybridization of forming the foregoing nucleotide sequence composition.
The invention provides a kind of protein antibodies, comprise polyclonal antibody and monoclonal antibody.Be easy to make polyclone and monoclonal antibody with method well known in the art at tumor suppressor gene encoded protein of the present invention (polypeptide) or its segment and analogue.In case purifying, these antibody can be used as laboratory reagent and diagnostic reagent with tumor suppressor gene diseases associated of the present invention.Gained antibody can be used for preparing antibody column, immunoprecipitation and identifies antigen with the Western trace.As follows with the milligram macro preparation at the universal method of the proteic monoclonal antibody of KIAA0157 coded by said gene of the present invention: with rabbit, horse, mouse and cavy as being subjected to immune animal, with a kind of scheme antigen inoculation albumen of immune animal known in the art, from the serum of collecting, separate (Fig. 1) such as IgG then.The also available ordinary method of monoclonal antibody of the present invention makes: carry out immunity with antigen protein inoculation mouse, take out spleen in the mouse body that shows enough antibody titerss.Separating Morr. cell makes the myeloma cell in selected B cell and B cell rice source merge the hybridoma that forms secretory antibody.Obtain hybridoma excretory monoclonal antibody with methods such as affinity column, ion-exchange or gel-filtration purifying from substratum.
The invention provides a kind of recombinant viral vector, it comprises forms the DNA that aforementioned a kind of nucleotide sequence is formed.
The invention provides a kind of dna fragmentation, this fragment can be used as primer, and is made up of the partial sequence of aforementioned a kind of nucleotide sequence.
The invention provides a kind of human diagnostic preparation, it comprises aforementioned probe and aforementioned proteinic antibody.Can be used for the inspection of genetic expression in the malignant tumours such as liver cancer, lung cancer, colorectal carcinoma, mammary cancer, leukemia, kidney, nervous system neoplasm.
The present invention also provides and has contained aforementioned proteic anti-tumor medicinal preparation.
Therefore, constitute the DNA of gene of the present invention, the protein of their transcript mRNA and translation thereof can be used as treatment, diagnosis and the prophylactic of kinds of tumors.
The present invention is described in conjunction with the accompanying drawings better:
The full cell distribution of Fig. 1 KIAA0157.KIAA0157-GFP rotaring copolymering NIH 3 T 3 cell, fluorescent microscope are observed the fluorescent signal distribution situation down.The result shows that KIAA0157 is full cell distribution.
The preparation of Fig. 2 KIAA0157 antibody.KIAA0157 (151-311aa)-GST carrier Transformed E .coliBL21 (DE3) bacterial strain, and purifying protein (A, B).(C) the KIAA0157 antiserum(antisera) detects the expression of KIAA0157 in HEK293.Serum titer is can detect the expression of target protein at 1: 500.
Fig. 3 utilizes the expression of tumor tissues chip detection KIAA0157.Detect the expression of KIAA0157 by hybridizing method at 19 kinds of cancers and corresponding adjacent tissues and 9 kinds of tumor cell lines.
Fig. 4 Real-time PCR detects the expression of KIAA0157 in liver cancer tissue and cancer beside organism.The result shows, has 10 routine patients' the expression of KIAA0157 in liver cancer tissue to be starkly lower than cancer beside organism in the 12 routine patients that detected.
Fig. 5 KIAA0157 suppresses cell proliferation.Behind the different tumour cells of PcDNA3.1-KIAA0157 or the transfection of PcDNA3.1 empty carrier, inoculation 6 orifice plates cultivated for 2 weeks after 12 hours, detected colony number of cell.The result shows that in three kinds of tumour cells, KIAA0157 crosses and expresses propagation capable of inhibiting cell.
Fig. 6 KIAA0157 expresses and to cause that the SKNSH cell G0/G1 phase blocks.KIAA0157-GFP or GFP empty carrier be transfection SKNSH cell respectively, utilizes Flow cytometry GFP to express and the cell cycle after 36 hours.
The acquisition of embodiment 1 people KIAA0157 cDNA
With the THAP11 gene is bait, utilize the general yeast-two hybrid technique in this area to study its interaction protein, obtain the section of DNA sequence, adopt this dna sequence dna to do the BLAST retrieval, the present invention finds to have higher homology with the Human genome KIAA0157 that infers.The total RNA of exhibitor medullary cell carries out reverse transcription by reverse transcription test kit (Promega company) specification sheets.Get the RNA of 1 μ g after quantitatively, add water and mend to 10.5 μ l, 70 ℃, 10min add 10 * buffer of 2 μ l, the MgCl of 2 μ l
2, 2 μ l the AMV (15U) of rRNasin, 1.5 μ l of oligo dT, 1 μ l of dNTP, 1 μ l, reaction system totally 20 μ l; 42 ℃, 1h; 94 ℃, 5min; 4 ℃, 5min.RNA is that reverse transcription is cDNA.With 5 '-CGGAATTCAATGTCCTACAG AGAGCAGGTT-3 '; 5 '-GGGGATCCTTAAATCTGGGAGG TCTGAGTG-3 ' is primer PCR amplification purpose fragment.The PCR condition is:
Cycle?1(1x)
94℃?5min
Cycle?2(30x)
94℃?1min
55℃?30s
72℃?1min
Cycle?3(1x)
72℃?10min
The PCR product carries out 1% agarose gel electrophoresis, the result produces special DNA band, and this dna fragmentation is connected with PMD-18-T carrier (TAKARA biotech firm), connects product Transformed E .coli JM109, after bacterium colony PCR is accredited as positive colony, order-checking after enzyme is cut evaluation again.Sequence after the order-checking is shown in the sequence 1.Because dna sequence dna is confirmed its verity, therefore, can produce the specific hybrid probe with following method: make the coli strain growth of carrying plasmid, plasmid DNA purification digests it with suitable Restriction Enzyme, the electrophoretic separation dna fragmentation.
Bioinformatic analysis shows that KIAA0157 protein has a coil-coil structural domain, and mediating protein interacts usually.Homology comparison shows that people and the proteic homology of mouse KIAA0157 are 93.5%, with the proteic homology of rat KIAA0157 be 94.2%.KIAA0157 albumen has so high homology explanation KIAA0157 gene in people, mouse and rat conservative at evolution of long period of time process camber, points out this gene may have important biological function.
Embodiment 2KIAA0157 Subcellular Localization
For observing the Subcellular Localization of KIAA0157, we form the KIAA0157-GFP integrative gene expression vector with the EcoRI/KpnI site that KIAA0157 cDNA is building up to the pEGFP carrier.With this expression vector rotaring copolymering NIH 3 T 3 cell, laser scanning co-focusing microscope detects and shows that fluorescent signal is distributed as full cell (Fig. 1).
The preparation of the proteic prokaryotic expression of embodiment 3KIAA0157, purifying and polyclonal antiserum
With the KIAA0157-Flag plasmid is template, with 5 '-CGGAATTCCCAAGGAACAAGAAAGAAGATT-3 '; 5 '-CCGCTCGAGTTAAATCTGGGAGG TCTGAGT-3 ' is a primer, utilizes 161 amino acid of C end of round pcr amplification KIAA0157.The PCR product is connected behind EcoRI, Xholl double digestion with the PGEX-4T-2 carrier, Transformed E .coliBL21 (DE3) competence bacterial strain, and the picking mono-clonal, after enzyme was cut evaluation, sequential analysis was correct.Conversion is had E.coliBL21 (DE3) the inoculation LB substratum of GST-KIAA0157 plasmid, and 37 ℃ are cultured to OD:0.6～1.0, and the IPTG, 30 ℃ that add 1mM induce 4h, centrifugal collection bacterium, and SDS-PAGE detects EDAG protein expression situation.Obtain purer GST-KIAA0157 fusion rotein through B-PER GST Spin Purification Kit purifying.The big ear of GST-KIAA0157 fusion protein immunization New Zealand with purifying is white, through behind the booster immunization several times, get rabbit anteserum, Western blot detects the sero-fast effect of KIAA0157, see Fig. 2, antiserum(antisera) is 1: 500 when dilution, can detect the target protein in the HEK29 cell of transfection KIAA0157-Myc plasmid.
Embodiment 4KIAA0157 has notable difference in the expression of multiple cancer and corresponding adjacent tissues
With 5 '-CGGAATTCAATGTCCTACAGAGAGCAGGTT-3 '; 5 '-GGGGATCCTTAAATCTGGGAGGTCTGAGTG-3 ' is a primer, adopts round pcr amplification cDNA segment as probe template, α-
32With CancerProfiling ArrayII hybridization, detect its expression behind the p mark at 19 kinds of cancers and corresponding adjacent tissues and 9 kinds of tumor cell lines.154 couples of cDNA that detected come from 154 patients' cancer and corresponding adjacent tissues.Results of hybridization shows that KIAA0157 all has expression in multiple tissue, and in different tumor tissues expression widely different (Fig. 3).In Tiroidina (8/10), liver (3/3), skin (6/10), breast (2/10), rectum (3/10), testis cancerous tissues such as (5/10), KIAA0157 expresses and significantly is lower than corresponding cancer beside organism; Then opposite in the middle of stomach-tissue (7/10), ovary tissue (6/10), prostata tissue (3/4), KIAA0157 expression level showed increased.Above result shows that the KIAA0157 expression does not have tissue specificity, and its function performance may be different with its tissue distribution relevant.In addition, in the lower right of film, arranged the cDNA of 9 kinds of tumor cell lines, KIAA0157 is expressed in all 9 kinds of tumor cell lines, wherein expression level is very high in HL60, A549, MLOT4, SW480 and the Raji clone, and the hybridization signal of other tumor cell line relatively a little less than.
We have further detected the expression level of KIAA0157 in the hepatocarcinoma patient.By liver cancer and the cancer beside organism total RNA of ordinary method extraction from 12 routine hepatocarcinoma patients.Obtain the first chain cDNA through reverse transcription.With 5 ' AGGAAATACTAGCCAGCAAGAG-3 '; 5 ' AAGATTCAACAACTCGCTCACT-3 ' are primer, utilize SYBRgreen reagent, carry out Real-time PCR.The PCR condition:
Cycle?1：(1X)
Step?1： 95.0℃ for?01:00.
Cycle?2：(40X)
Step?1： 95.0℃ for?00:10.
Step?2： 65.0℃ for?00:30.
Cycle?3：(1X)
Step?1： 95.0℃ for?01:00.
Cycle?4：(1X)
Step?1： 55.0℃ for?01:00.
Cycle?5：(81X)
Step?1： 55.0℃-95.0℃?for?00:10.
Use IQ
TM5 Multicolor Real-time PCR Detection System monitor in real time to report fluorescence dye institute emitted fluorescence.The fluorescent emission amount reflects cycle number and is read by the software of sequential detection instrument that provide the significant cycle number threshold value of pcr amplification, the value of cycle number and genomic dna logarithm are linear.The result shows (Fig. 4), has in 10 routine patients' the liver cancer tissue KIAA0157 express in the 12 routine hepatocarcinoma patients and is starkly lower than cancer beside organism, shows KIAA0157 down-regulated expression in liver cancer takes place.
Embodiment 5KIAA0157 crosses the expression inhibiting cell colony and forms
Whether on cell proliferation is influential in order to study KIAA0157, and we have made up KIAA0157-myc/hisB
+Carrier for expression of eukaryon, and adopt colony formation experiment to detect KIAA0157 and in the SKNSH cell, cross the changing conditions of expressing its propagation of back.As shown in Figure 5, transfection KIAA0157-myc/hisB
+The SKNSH cell with respect to the cell that changes empty carrier, the ability that its colony forms is obviously suppressed.In liver cancer cell HepG2 cell and breast cancer cell MCF-7 cell, also obtain identical result.
Embodiment 6KIAA0157 expresses and to cause that the SKNSH cell G0/G1 phase blocks
Be to understand KIAA0157 to the influence of SKNSH cell cycle, we at the SKNSH cell transfecting carrier for expression of eukaryon of KIAA0157 of band GFP label, flow cytometer detect the cell cycle each the time phase distribution.Compare with GFP-transfected empty carrier, the SKNSH cell the tangible G0/G1 phase occurs and blocks (Fig. 6) behind the transfection KIAA0157 carrier for expression of eukaryon.
Sequence table
＜110〉INST OF EMISSION ﹠ RADIATION M
＜120〉a kind of new human tumor inhibiting gene KIAA 0157 and application thereof
<160>2
<210>1
<211>1248
<212>DNA
＜213〉ethnic group (Homo Sapiens.)
<400>1
atggcggcgt?ccatttcggg?ctacaccttc?agtgctgtgt?gtttccacag?cgccaacagc 60
aacgcggacc?acgaaggatt?tttactggga?gaggtaagac?aagaggaaac?gtttagcatc 120
agtgactcac?aaatcagcaa?cacagaattt?ctgcaagtaa?ttgaaatcca?taaccatcag 180
ccttgttcaa?aactttttag?tttttatgac?tacgcaagca?aagtgaatga?ggagagtttg 240
gacaggattc?ttaaagatcg?gagaaagaaa?gtcattgggt?ggtacagatt?ccggcgcaat 300
acgcagcagc?agatgtccta?cagagagcag?gttcttcaca?agcagctcac?ccgcatcctc 360
ggcgtgcccg?acctcgtctt?tcttctcttc?agcttcatct?ccactgccaa?caattccact 420
cacgctttag?aatatgtgct?cttcagacca?aatagaaggt?ataatcagag?gatatcactc 480
gctattccca?atctaggaaa?tactagccag?caagagtaca?aagtgtcttc?agtgccaaat 540
acttctcaga?gttatgccaa?agtgattaaa?gaacatggta?ctgacttttt?tgacaaggat 600
ggagtgatga?aagacatcag?ggcgatttat?caggtttata?atgcacttca?ggagaaagtt 660
caggcagtgt?gtgcagatgt?tgaaaagagt?gagcgagttg?ttgaatcttg?tcaggcagaa 720
gtgaacaaat?taagaagaca?aatcactcag?aggaaaaatg?aaaaggaaca?agaaagaaga 780
ttgcagcagg?cagtgttaag?cagacagatg?ccgtctgaaa?gcttggaccc?agcgttcagt 840
cctcggatgc?cgtcctctgg?gtttgcagct?gaaggcagaa?gtacacttgg?agatgcagag 900
gcctcggatc?ctcctccccc?ttactctgat?tttcacccaa?acaatcaaga?aagtactttg 960
agccactctc?gcatggaaag?gagtgtcttt?atgcctcgac?ctcaagctgt?gggctcttcc?1020
aattatgctt?ccaccagtgc?cggactgaag?tatcctggaa?gtggggctga?ccttcctcct?1080
ccccaaagag?cagctggaga?cagtggtgag?gattcagacg?acagtgatta?tgaaaatttg 1140
attgacccta?cagagccttc?taatagtgaa?tactcacatt?caaaggattc?tcgacccatg 1200
gcacatcccg?acgaggaccc?caggaacact?cagacctccc?agatttaa 1248
<210>2
<211>416
<212>PRT
＜213〉ethnic group (Homo Sapiens.)
<400>2
Met?Ala?Ala?Ser?Ile?Ser?Gly?Tyr?Thr?Phe?Ser?Ala?Val?Cys?Phe?His 16
Ser?Ala?Asn?Ser?Asn?Ala?Asp?His?Glu?Gly?Phe?Leu?Leu?Gly?Glu?Val 32
Arg?Gln?Glu?Glu?Thr?Phe?Ser?Ile?Ser?Asp?Ser?Gln?Ile?Ser?Asn?Thr 48
Glu?Phe?Leu?Gln?Val?Ile?Glu?Ile?His?Asn?His?Gln?Pro?Cys?Ser?Lys 64
Leu?Phe?Ser?Phe?Tyr?Asp?Tyr?Ala?Ser?Lys?Val?Asn?Glu?Glu?Ser?Leu 80
Asp?Arg?Ile?Leu?Lys?Asp?Arg?Arg?Lys?Lys?Val?Ile?Gly?Trp?Tyr?Arg 96
Phe?Arg?Arg?Asn?Thr?Gln?Gln?Gln?Met?Ser?Tyr?Arg?Glu?Gln?Val?Leu 112
His?Lys?Gln?Leu?Thr?Arg?Ile?Leu?Gly?Val?Pro?Asp?Leu?Val?Phe?Leu 128
Leu?Phe?Ser?Phe?Ile?Ser?Thr?Ala?Asn?Asn?Ser?Thr?His?Ala?Leu?Glu 144
Tyr?Val?Leu?Phe?Arg?Pro?Asn?Arg?Arg?Tyr?Asn?Gln?Arg?Ile?Ser?Leu 160
Ala?Ile?Pro?Asn?Leu?Gly?Asn?Thr?Ser?Gln?Gln?Glu?Tyr?Lys?Val?Ser 176
Ser?Val?Pro?Asn?Thr?Ser?Gln?Ser?Tyr?Ala?Lys?Val?Ile?Lys?Glu?His 192
Gly?Thr?Asp?Phe?Phe?Asp?Lys?Asp?Gly?Val?Met?Lys?Asp?Ile?Arg?Ala 208
Ile?Tyr?Gln?Val?Tyr?Asn?Ala?Leu?Gln?Glu?Lys?Val?Gln?Ala?Val?Cys 224
Ala?Asp?Val?Glu?Lys?Ser?Glu?Arg?Val?Val?Glu?Ser?Cys?Gln?Ala?Glu 240
Val?Asn?Lys?Leu?Arg?Arg?Gln?Ile?Thr?Gln?Arg?Lys?Asn?Glu?Lys?Glu 256
Gln?Glu?Arg?Arg?Leu?Gln?Gln?Ala?Val?Leu?Ser?Arg?Gln?Met?Pro?Ser 272
Glu?Ser?Leu?Asp?Pro?Ala?Phe?Ser?Pro?Arg?Met?Pro?Ser?Ser?Gly?Phe 288
Ala?Ala?Glu?Gly?Arg?Ser?Thr?Leu?Gly?Asp?Ala?Glu?Ala?Ser?Asp?Pro 304
Pro?Pro?Pro?Tyr?Ser?Asp?Phe?His?Pro?Asn?Asn?Gln?Glu?Ser?Thr?Leu 320
Ser?His?Ser?Arg?Met?Glu?Arg?Ser?Val?Phe?Met?Pro?Arg?Pro?Gln?Ala 336
Val?Gly?Ser?Ser?Asn?Tyr?Ala?Ser?Thr?Ser?Ala?Gly?Leu?Lys?Tyr?Pro 352
Gly?Ser?Gly?Ala?Asp?Leu?Pro?Pro?Pro?Gln?Arg?Ala?Ala?Gly?Asp?Ser 368
Gly?Glu?Asp?Ser?Asp?Asp?Ser?Asp?Tyr?Glu?Asn?Leu?Ile?Asp?Pro?Thr 384
Glu?Pro?Ser?Asn?Ser?Glu?Tyr?Ser?His?Ser?Lys?Asp?Ser?Arg?Pro?Met 400
Ala?His?Pro?Asp?Glu?Asp?Pro?Arg?Asn?Thr?Gln?Thr?Ser?Gln?Ile 415
Claims (10)
1. DNA who constitutes people's gene has nucleotide sequence shown in the sequence 1 or shows the nucleotide sequence of one or more base deletions, replacement or increase with respect to described nucleotides sequence.
2. protein, the aminoacid sequence that it has aminoacid sequence shown in the sequence 2 or one or more aminoacid deletion is arranged, substitute or increase with respect to described aminoacid sequence.
3. probe, it is contained in the DNA of the DNA hybridization that the described nucleotide sequence of claim 1 forms.
4. recombinant viral vector, it comprises the DNA that is made up of the described nucleotide sequence of claim 1.
5. the dna fragmentation as primer is made up of the partial sequence of the described nucleotide sequence of claim 1.
6. human diagnostic preparation, it comprises the described probe of claim 3.
7. the described diagnostic preparation of claim 6 application that genetic expression is checked in malignant tumours such as liver cancer, lung cancer, colorectal carcinoma, mammary cancer, leukemia, kidney, nervous system neoplasm.
8. anti-tumor medicinal preparation, it contains the described protein of one of claim 2.
9. an energy and claim 2 described protein specific bonded polyclone and monoclonal antibody.
10. diagnostic pharmaceutical preparation, it contains the expression that the described antibody of claim 9 is used for detecting oncogene.
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| CN106432465A (en) * | 2016-11-08 | 2017-02-22 | 李露青 | Human tumor-inhibiting protein variant and application thereof |
| CN106397567A (en) * | 2016-11-08 | 2017-02-15 | 李露青 | Human-tumor inhibition protein variants and application thereof |
| CN106349360A (en) * | 2016-11-09 | 2017-01-25 | 李露青 | Human tumor suppression protein variant and application thereof |
| CN106349362A (en) * | 2016-11-09 | 2017-01-25 | 李露青 | Human tumor suppression protein variant and application thereof |
| CN106349361A (en) * | 2016-11-09 | 2017-01-25 | 李露青 | Human tumor suppression protein variant and application thereof |
| CN106349364A (en) * | 2016-11-10 | 2017-01-25 | 李露青 | Human tumor suppression protein variant and application thereof |
| CN106349363A (en) * | 2016-11-10 | 2017-01-25 | 李露青 | Human tumor suppression protein variant and application thereof |
| CN106432466A (en) * | 2016-11-10 | 2017-02-22 | 李露青 | Human tumor arrestin variant and application thereof |
| CN106349365A (en) * | 2016-11-14 | 2017-01-25 | 李露青 | Human tumor suppression protein variant and application thereof |
| CN106349366A (en) * | 2016-11-14 | 2017-01-25 | 李露青 | Human tumor suppression protein variant and application thereof |
| CN106478799A (en) * | 2016-11-14 | 2017-03-08 | 李露青 | A kind of human tumor inhibiting protein variant and its application |
| CN107723368A (en) * | 2017-11-28 | 2018-02-23 | 杭州可帮基因科技有限公司 | One group of gene for being used for clear-cell carcinoma molecule parting and its application |
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