CN104672317B - Adjust transcription factor ZNF312b and its application of Oct4 gene expressions - Google Patents
Adjust transcription factor ZNF312b and its application of Oct4 gene expressions Download PDFInfo
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- CN104672317B CN104672317B CN201510075746.3A CN201510075746A CN104672317B CN 104672317 B CN104672317 B CN 104672317B CN 201510075746 A CN201510075746 A CN 201510075746A CN 104672317 B CN104672317 B CN 104672317B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
Abstract
The present invention discloses transcription factor ZNF312b and its application of a kind of regulation Oct4 gene expressions.Transcription factor ZNF312b amino acid sequence is as shown in SEQ ID No.1.Transcription factor ZNF312b nucleotide sequence is encoded as shown in SEQ ID No.2.Present invention firstly discovers that transcription factor ZNF312b can activate key transcription factor Oct4 genes in MSCs, promote MSCs propagation.Therefore, the transcription factor can be used to prepare regulation Oct4 gene expressions preparation and/or prepare to promote MSCs propagation preparations.
Description
Technical field
The present invention relates to genetic engineering and biology field, and in particular to a kind of transcription of regulation Oct4 gene expressions
Factor Z NF312b and its application.
Background technology
Mescenchymal stem cell (mesenchymal stem cells, MSCs) is that one kind being capable of self-renewing and with multidirectional
The stem cell of differentiation potential, it can break up to histocytes such as fat, bone, cartilage, nerve, muscle and liver cells, so in group
The fields such as weaver's journey, cell transplantation and gene therapy have huge application potential.MSCs is not only the important of body microenvironment
Part, and the effect of secretion cytokine profiles, growth factor and substrate molecule is respectively provided with vivo and in vitro.MSCs can table
Up to embryonic stem cell marker Oct4, Nanog, alkaline phosphatase and SSEA-4.But MSCs maintain self-renewing with it is undifferentiated
The molecular mechanism of state is also unclear.
Oct4 belongs to the member of POU transcription factor families, is produced by pou5f1 gene codes.Oct4 is to participate in regulation and control embryo
The self-renewing of stem cell (embryonic stem cells, ESCs), maintains the important transcription factor of its totipotency, is recognized
For the important symbol thing of stem cell multi-lineage potential.ZNF312b genes are also referred to as forebrain embryo's zinc finger sample protein I
(Fezfl) gene, be Africa xenopus (Xenopus) forebrain Olfactory sensory neurons (OSN) in first certified gene, and
And known coded has the albumen of zinc ion.Appropriate termination (the proper termination of of the gene pairs olfactory nerve
Theolfactory nerve), the formation of olfactory bulb and the towering progenitor cells of intermediate nerve through being essential for head end migration stream
, and axon guidance (axonal projection) cell self-contr ol and olfactory bulb film are formed for control and important
Factor.It is known to have a small amount of report on ZNF312b gene functions, but they are relevant with the development of stomach cancer.There is research
Show, ZNF312b overexpression is detected in the stomach organization and stomach cancer cell line of people, the ZNF312b mistake in nude mouse
Expression can induced cancer formed, ZNF312b can be as transcription factor induced tumor gene K-ras expression, so as to activate
The ERK signal transduction pathway relevant with cell propagation, causes the formation of stomach cancer；In stomach cancer cell, DNA demethylations and group egg
Baiyi, which is acylated, promotes Sp1 transcription factors and ZNF312b promoter results, so as to activate ZNF312b expression.So far there are no document
Report the adjustment effect expressed about ZNF312b Oct4.
The content of the invention
The shortcomings that primary and foremost purpose of the present invention is to overcome prior art and deficiency, there is provided one kind regulation Oct4 gene expressions
Transcription factor ZNF312b.
Another object of the present invention is to the application for the transcription factor ZNF312b for providing the regulation Oct4 gene expressions.
The purpose of the present invention is achieved through the following technical solutions：A kind of transcription factor of regulation Oct4 gene expressions
ZNF312b, its amino acid sequence is as shown in SEQ ID No.1.
The transcription factor ZNF312b of described regulation Oct4 gene expressions, its coding nucleotide sequence such as SEQ ID No.2
It is shown.
A kind of carrier for recombinantly expressing transcription factor ZNF312b, is by expression vector and above-mentioned regulation Oct4 gene expressions
Transcription factor ZNF312b coding nucleotide sequence recombinate to obtain.
Described expression vector is prokaryotic expression carrier or carrier for expression of eukaryon, preferably carrier for expression of eukaryon.
A kind of recombinant bacterium that can express transcription factor ZNF312b, it is by above-mentioned recombination expression transcription factor ZNF312b load
Body is transferred to engineering bacteria and obtained.
A kind of recombinant cell that can express transcription factor ZNF312b, it is by above-mentioned recombination expression transcription factor ZNF312b
Carrier is transferred to engineering cell and obtained.
The transcription factor ZNF312b of described regulation Oct4 gene expressions prepare regulation Oct4 gene expressions preparation and/
Or prepare the application promoted in MSCs propagation preparations.
Described preparation is preferably medicine.
The transcription factor ZNF312b of described regulation Oct4 gene expressions prepare regulation Oct4 gene expressions preparation and/
Or the application promoted in MSCs propagation preparations is prepared, preferably include following steps：
(1) the transcription factor ZNF312b of described regulation Oct4 gene expressions coding nucleotide sequence and expression are carried
Body weight group, obtain recombinantly expressing ZNF312b carrier；
(2) carrier for recombinantly expressing ZNF312b is transferred in host cell and expressed, purified, adjusted Oct4 bases
Because of the transcription factor ZNF312b of expression；
The regulation Oct4 gene tables that the carrier for the recombination expression ZNF312b that step (1) obtains obtains after purification with step (2)
The transcription factor ZNF312b reached is regulation Oct4 gene expressions preparation and/or promotes MSCs propagation preparations.
The present invention is had the following advantages relative to prior art and effect：
Contain enhancer, including Site 2A and Site 2B in the upstream noncoding region of Oct4 genes.Present invention firstly discovers that
Transcription factor ZNF312b is in MSCs to the regulation mechanism of Oct4 gene expressions.The present invention is using the success of yeast one-hybrid technology
The transcription factor ZNF312b with Site 2A specific bindings is cloned in ground, it was demonstrated that the transcription factor ZNF312b energy in MSCs
Enough and Oct4 genes Site 2A are specifically bound, and can activate the transcription of Oct4 genes.Transcription factor ZNF312b can not
Combined with Site 2B, Site 2B do not work to activation Oct4 gene expressions.The present invention is also instantaneous by ZNF312b expression plasmids
Transfection is expressed into MSCs, it is found that ZNF312b can promote MSCs propagation.Present invention is disclosed transcription factor ZNF312b
To adjust key transcription factor Oct4 genes molecular regulation mechanism, be self-renewing of the ZNF312b transcription factors to MSCs and
Multi-lineage potential plays crucial adjustment effect and provides theoretical foundation.
Brief description of the drawings
Fig. 1 is to be related to carry out the Vector map figure that yeast one-hybrid technology uses in the embodiment of the present invention 1, wherein scheming
A is the pAbAi carriers for building bait plasmid, and figure B is the pAD-GAL4 carriers for construction expression plasmid.
Fig. 2 is the expression inspection that BM-MSCs is divided into Oct4 and ZNF312b after variety classes cell in the embodiment of the present invention 2
Survey result figure.
Fig. 3 is that reporter gene luciferase plasmids and different promoters are built into plasmid simultaneously in the embodiment of the present invention 3
It is intracellular to be transfected into undifferentiated BM-MSCs and HeLa, and reporter gene luciferase plasmids, different promoters are built
The result figure detected after plasmid and ZNF312b expression plasmids while transfection HeLa cell.
Fig. 4 is by Oct4, ZNF312b, Oct4siRNA and ZNF312b siRNA expression plasmids point in the embodiment of the present invention 4
BM-MSCs proliferative conditions result figure after being expressed into BM-MSCs is not transiently transfected.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but is not limited to the model of the present invention
Enclose.If unspecified, embodiment is carried out with reference to conventional laboratory conditions, or with reference to the specification of kit manufacturer.
Used engineering bacteria, cell line are commercialized bacterial strain or cell line in embodiment.The system of competent escherichia coli cell
It is standby according to《Molecular cloning》Prepared.
The yeast one-hybrid technology screening of embodiment 1 and the transcription factor of Site 2A specific bindings
According to Clonetech companies yeast one-hybrid kit Matchmaker Gold Yeast One-Hybrid
The operation requirement of Library Screening System specifications, builds bait plasmid and bait yeast strain.In order to improve
The discrimination and Percentage bound of transcription factor and cis-acting elements, it is substantially single with the Site2A nucleotides sequences row of OCT4 genes
Position, and the Xho I (ctcgag) and Hind III to match with Yeast genome integration vector pAbAi (Figure 1A) is introduced at both ends
(aagctt) restriction enzyme site, 3 repetition Site 2A sequences containing Xho I and Hind III digestions site finally are being synthesized just
After adopted chain and antisense strand (as shown in SEQ ID No.3 and 4), positive-sense strand and antisense strand mix with molal quantity, after being heated to 95 DEG C
Naturally anneal, obtain triple Site 2A oligonucleotide chains with duplex structure.It is double respectively with Xho I and Hind III again
The triple Site 2A oligonucleotides chains of digestion and pAbAi carriers, it is thin that connection product is converted into Escherichia coli 10 competence of Top
Born of the same parents, with the LB solid medium screening positive clones of the benzyl mycin of ammonia containing 100mg/L, extract positive colony plasmid, through Xho I and
Developed the color and identified with 12% PAGE and silver staining after Hind III double digestions, the insertion of verifying purpose fragment, take endonuclease bamhi length
Correct recombinant plasmid is sequenced, and identifies that insetion sequence and triple Site 2A oligonucleotide chain-orderings are completely the same and square
To correct, illustrate that bait carrier pSite2A-AbAi is successfully constructed.The μ g of pSite2A-AbAi bait carriers 1 are taken, through BstB I enzymes
Transformed yeast Y1HGold competent cells after tangent linearization, conversion fluid are coated with SD/-Ura solid mediums, and Dan Ke is chosen after 3 days
It is grand to carry out bacterium colony PCR identifications with Matchmaker Insert Check PCR Mix 1, it was demonstrated that the pSite2A-AbAi of linearisation
Bait carrier has been incorporated into yeast Y1HGold genomes, so as to successfully obtain bait yeast.
Then, yeast one-hybrid library is built.With TRIzol methods extraction BM-MSCs (HUXMA-01001, it is (wide purchased from match industry
State) bio tech ltd) total serum IgE, gel electrophoresis analysis prove RNA without degraded and quality it is good.It is mould to take 1 μ g total serum IgEs
Plate, with the synthesizing single-stranded cDNA of SMART technologies, LD-PCR is carried out as template using single-stranded cDNA and expands synthetic double chain cDNA, is used in combination
CHROMA SPINTE-400 resin chromatographies post purifies.By the double-strand cDNA of purifying and the pAD-GAL4 expression vectors of linearisation (figure
1B) it is attached, converts escherichia coli jm109 competent cell, obtain cDNA library.
Finally, the transcription factor of screening and Site 2A specific bindings is carried out.CDNA library conversion is contained into pSite2A-
AbAi bait competent yeast cells, it is coated on the screening that cDNA library is carried out on SD/-leu/AbA flat boards.Conversion fluid dilutes
211 clones are grown after to 1/100 on SD/-leu/AbA flat boards.Total clone's number according to formula below calculating sifting：Total gram
Grand number=cfu/ applies plate bulk × dilution rate × cell suspension cumulative volume.Result is that total clone's number is 3.17 × 106, cDNA library
Recombination fraction and transformation efficiency meet the requirements.Picking above monoclonal, NCBI Blast are carried out after sequencing and compare analysis, knot
Fruit finds to have screened ZNF312b transcription factors (SEQ ID No.1-2).This illustrates that this laboratory uses yeast one-hybrid skill
Art has successfully cloned the transcription factor ZNF312b with Site 2A specific bindings.
Embodiment 2 detects expression of the Oct4 and ZNF312b genes in BM-MSCs
Respectively out of undifferentiated BM-MSCs (0 day), or by BM-MSCs Osteoblast Differentiation, into cartilage differentiation and into fat
Differentiation (interstital stem cell skeletonization, is purchased from the match limited public affairs of industry biotechnology in Guangzhou into cartilage, Adipogenic induction differential medium
Department) 7 days afterwards, the total mRNA of intracellular extraction of 14 days and 21 days, reverse transcription cDNA, then utilize the forward and reverse of target gene
Primer is expanded to detect its expression, and target gene has respectively：GAPDH (primer sequences：SEQ ID No.5-6),
Oct4 (primer sequences：SEQ ID No.7-8) and ZNF312b (primer sequences：SEQ ID No.9-10).PCR reaction system
For：Each 1 μ l, 10mmol/L dNTP mix 0.4 μ l, 0.5U/ μ of the forward and reverse primers of μ l, 10pmol/ μ l of 10-20ng/ μ l templates 1
The μ l of 1 μ l, 10 × PCR reaction buffer of l high-fidelity Taq archaeal dna polymerases 2, remaining is water, totally 20 μ l.PCR reaction condition：94
DEG C 5 minutes；94 DEG C 20 seconds, 55 DEG C 20 seconds, 72 DEG C 1 minute, totally 35 circulation；72 DEG C 10 minutes.
As a result show, undifferentiated BM-MSCs (0 day), 7 days after Osteoblast Differentiation, or 7 days after cartilage differentiation, Adipose Differentiation
The cell of 7 days can express Oct4 and ZNF312b mRNA afterwards, and the cell of 14 days and 21 days can not all express after three kinds of differentiation
Oct4 and ZNF312b mRNA (Fig. 2).
The luciferase reporter gene of embodiment 3 is analyzed
It is respectively synthesized Oct4Site 2A (the SEQ ID containing BstBI (ttcgaa) and BbsI (tctcta) restriction enzyme site
No.11), Site 2B gene orders (SEQ ID No.12), Site 2A+2B gene orders (SEQ ID No.13) and Site
2A+2A gene orders (SEQ ID No.14), describe for convenience, these sequences are referred to as Oct4 promoter DNAs.By Oct4
Promoter DNA and pGL-3basic plasmids (Promega) carry out BstBI and BbsI double digestions respectively, and digestion condition difference is as follows：
10 μ l or Oct4 promoter DNA of pGL3-basic plasmids 30 μ l, BstBI (10U/ μ L) 2 μ l, BbsI (10U/ μ l) 2 μ l, 10 ×
Buffer 5 μ l, ddH2O complements to 50 μ l.After 37 DEG C of digestions overnight, in 2% agarose gel electrophoresis, and digestion is separately recovered
Oct4 promoter DNAs and pGL3-basic plasmids afterwards.The Oct4 promoter DNA double digestion products of recovery are connected to pGL3-
In basic plasmids, and the pGL3-basi-Oct4 promoter plasmids of this restructuring are converted into bacillus coli DH 5 alpha competent cell, obtained
DNA pGL3-basic DH5 α recombinant clones that must be containing Oct4 promoters.Specific method is as follows：Oct4 promoters are taken respectively
6 μ l, pGL3-basic plasmid double digestion recovery products (160ng/ μ l) of DNA double digestion recovery product (68ng/ μ l) 2 μ l, 10 ×
2 μ l, T4DNA ligase (5U/ μ l, Fermentas) of buffer 1 μ l, ddH2O9 μ l, totally 20 μ l, 16 DEG C of connection 2h, by 20 μ l
Connection product and the competent cells of 100 μ l Escherichia coli DH α 5 mix, and place 30min, 42 DEG C of 90s on ice, on ice 5min, then
700 μ l LB non-resistant culture mediums are added, after 37 DEG C of 250r/min cultivate 1h, are applied to the solid training of the resistances of Amp containing 100mg/L
Support base flat board, 37 DEG C of overnight incubations, there is clone to grow on flat board, some clones of picking carry out positive identifications, select positive colony
In Shanghai, Sheng Gong companies are sequenced.Sequencing result shows that its nucleotide sequence is all correct, obtains pGL3-basic Site
2A, pGL3-basic Site 2B, pGL3-basic Site2A+2B, pGL3-basic Site 2A+2A promoter plasmids.
UseHD transfection reagents (Promega companies) are transfected, and transfection object is one group undifferentiated
BM-MSCs and two group of Human cervical cancer cell lines HeLa cell.For undifferentiated BM-MSCs and lineup's cervical cancer tumer line
HeLa cells, it is each separately transfected into pGL3-basic plasmids, pGL3-basic Site 2A, the pGL3-basic Site of structure
2B, pGL3-basic Site 2A+2B, pGL3-basic Site 2A+2A promoter plasmids, while it is transferred to pRLSV40 fluorescence
Plain expression of enzymes plasmid (E2231, Promega company) compares as transfection efficiency internal reference.For another group of Human cervical cancer cell lines
HeLa cells, it is each separately transfected into pGL3-basic plasmids, pGL3-basic Site 2A, the pGL3-basic Site of structure
2B, pGL3-basic Site 2A+2B, pGL3-basic Site 2A+2A promoter plasmids, while it is transferred to pRLSV40 fluorescence
Plain expression of enzymes plasmid and pGL3-basic the ZNF312b expression plasmid (transcription factor gene containing ZNF312b for obtaining embodiment 1
The clone of sequence and pGL3-basic deliver completes building process by Guangzhou Hui Jun bio tech ltd, and sequencing result shows
PGL- is inserted into by BstBI (ttcgaa) and BbsI (tctcta) restriction enzyme site for the sequence as shown in SEQ ID No.2
In 3basic plasmids).Transfectional cell is collected after 24h, (Passive Lysis Buffer, Promega are public with passive lysate
Department) cell pyrolysis liquid is made, use the Relative luciferase activity assay kit and GloMax 2/20 of Promega companies
Luminometer determines fluorescent value.As a result show, in undifferentiated BM-MSCs, Site 2A can promote luciferase table
Reach, intracellular in HeLa, ZNF312b can activate Site2A, and so as to promote luciferase expression, two Site 2A can promote
The expression of more luciferases；Intracellular in undifferentiated BM-MSCs and HeLa, Site 2B and ZNF312b can not promote fluorescence
Plain expression of enzymes (Fig. 3).As a result demonstrating ZNF312b transcription factors can be combined with Site 2A regions, not tied with Site 2B regions
Close.
Embodiment 4 detects the influence of ZNF312b transcription factors on B M-MSCs propagation
The primer of amplification ZNF312b and Oct4 gene open reading frames is separately designed with Primer 5.0：Expand Oct4's
Primer sequence expands ZNF312b primer sequence as shown in SEQ ID No.17-18, primer as shown in SEQ ID No.15-16
Both ends restriction enzyme site is respectively Xho I, Nco I.Primer is synthesized by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd.Amplification
ZNF312b genes and Oct4 genes are that with gastric carcinoma cell lines MKN28 and human embryonic stem cell H9, (being purchased from Jiangsu, this is smooth respectively
Good fortune Bioisystech Co., Ltd) cDNA be template, the reaction condition of PCR amplifications：94 DEG C 5 minutes；94 DEG C 20 seconds, 55 DEG C 20
Second, 72 DEG C 1 minute, totally 35 circulation；72 DEG C 10 minutes.PCR primer carries out 2% agarose gel electrophoresis, and PCR primer is returned
Receive purifying.The PCR primer of recovery is cloned into pGL3-Basic carrier for expression of eukaryon, be respectively designated as pGL3-ZNF312b and
PGL3-Oct4, DNA sequencing show that sequence is correct.Oct4siRNA(SEQ ID No.19)、ZNF312b siRNA(SEQ ID
No.20) siRNA (SEQ ID No.21) of nothing to do with is bought in Jin Da bio tech ltd of Beijing Cigna.By BM-
MSCs cells are inoculated in six orifice plates, are placed in 5%CO237 DEG C of incubators in, with the DMEM/F12 containing 10% hyclone
Culture medium is cultivated.When Monolayer growth of cells density reaches 80%~90%, pressHD transfection reagent boxes
(Promega companies) operating procedure transfected plasmids.Transfected according to following packet：Compare the BM-MSCs for untransfected, Oct4
To transiently transfect the BM-MSCs of pGL3-Oct4 plasmids, siOct4 is the BM-MSCs for transiently transfecting Oct4siRNA；SiOct4 pairs
According to being to transiently transfect unrelated siRNA as negative control；ZNF312b is the BM- for transiently transfecting pGL3-ZNF312b plasmids
MSCs, siZNF312b are the BM-MSCs for transiently transfecting ZNF312b siRNA, and siZNF312b controls are unrelated to transiently transfect
SiRNA is as negative control.
As a result show, Oct4 and ZNF312b can promote BM-MSCs propagation, Oct4siRNA and ZNF312b siRNA
BM-MSCs propagation can be suppressed, unrelated siRNA breeds without influence (Fig. 4) as negative control on BM-MSCs.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (3)
1. adjust the transcription factor ZNF312b of Oct4 gene expressions application, it is characterised in that：Described regulation Oct4 gene tables
The transcription factor ZNF312b reached answering in preparation regulation Oct4 gene expressions preparation and/or preparation promote MSCs propagation preparations
With；
The transcription factor ZNF312b of described regulation Oct4 gene expressions amino acid sequence is as shown in SEQ IDNo.1.
2. the transcription factor ZNF312b of regulation Oct4 gene expressions according to claim 1 application, it is characterised in that：
Described preparation is medicine.
3. the transcription factor ZNF312b of regulation Oct4 gene expressions according to claim 1 application, it is characterised in that bag
Include following steps：
(1) by the transcription factor ZNF312b of described regulation Oct4 gene expressions coding nucleotide sequence and expression vector weight
Group, obtain recombinantly expressing ZNF312b carrier；
(2) carrier for recombinantly expressing ZNF312b is transferred in host cell and expressed, purified, adjusted Oct4 genes table
The transcription factor ZNF312b reached；
The regulation Oct4 gene expressions that the carrier for the recombination expression ZNF312b that step (1) obtains obtains after purification with step (2)
Transcription factor ZNF312b is regulation Oct4 gene expressions preparation and/or promotes MSCs propagation preparations.
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| CN109913499A (en) * | 2017-12-13 | 2019-06-21 | 上海吉凯基因化学技术有限公司 | It is a kind of suitable for long-chain non-coding RNA construct and express integrated slow virus carrier system and its application |
| CN116790634B (en) * | 2023-06-19 | 2024-04-16 | 昆明理工大学 | Zinc finger transcription factor gene and application thereof |
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| KR100861465B1 (en) * | 2007-08-06 | 2008-10-02 | 한국생명공학연구원 | A gastric carcinoma gene znf312b, a protein translated from the gene and a diagnostic kit using the same |
| WO2009020346A3 (en) * | 2007-08-06 | 2009-04-16 | Korea Res Inst Of Bioscience | A gastric carcinoma gene znf312b, a protein translated from the gene, and a diagnostic kit and a screening method for anticancer agents using the same |
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|---|---|---|---|---|
| KR100861465B1 (en) * | 2007-08-06 | 2008-10-02 | 한국생명공학연구원 | A gastric carcinoma gene znf312b, a protein translated from the gene and a diagnostic kit using the same |
| WO2009020346A3 (en) * | 2007-08-06 | 2009-04-16 | Korea Res Inst Of Bioscience | A gastric carcinoma gene znf312b, a protein translated from the gene, and a diagnostic kit and a screening method for anticancer agents using the same |
| CN101688207A (en) * | 2007-08-06 | 2010-03-31 | 韩国生命工学研究院 | A gastric carcinoma gene znf312b, a protein translated from the gene, and a diagnostic kit and a screening method for anticancer agents using the same |
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| Title |
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| Human ZNF312b oncogene is regulated by Sp1 binding to its promoter region through DNA demethylation and histone acetylation in gastric cancer;In-Sung Song et al.,;《International Journal of Cancer》;20111101;第129卷(第9期);2124-2133 * |
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