CN1563380A - Gene oder of directivity and differentiation factor 1 fat cell of pigs - Google Patents
Gene oder of directivity and differentiation factor 1 fat cell of pigs Download PDFInfo
- Publication number
- CN1563380A CN1563380A CN 200410017860 CN200410017860A CN1563380A CN 1563380 A CN1563380 A CN 1563380A CN 200410017860 CN200410017860 CN 200410017860 CN 200410017860 A CN200410017860 A CN 200410017860A CN 1563380 A CN1563380 A CN 1563380A
- Authority
- CN
- China
- Prior art keywords
- sequence
- gene
- genbank
- add1
- differentiation factor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Peptides Or Proteins (AREA)
Abstract
A gene sequence of a pig fat cell orientation and differential factor 1 is used in gene engineering technological field Gone ADD1, its CDNA sequence and coded protein amino acid sequence amplified from the master RNA of pig's splenic organ have the total gene length of 3830bp, open reading frame 11bp-3453bp segments coded with 1151 amino acid protein, its 3' end un-coded sequence length is 10bp and 5' end is 366bp, a helix-loop-helix core structure is at 320-380 of the protein sequence which is used f in the research of said gene function in pig fat cells separation and its cholesterin level adjustment and plays an instructive role in the production and seeding of lean and low cholesterin by MAS method.
Description
Technical field
What the present invention relates to is a kind of albumen coded sequence that is used for gene engineering technology field, the gene order of particularly a kind of porcine adipocyte orientation and differentiation factor 1.
Technical background
Fatty character is that animal body grows and one of leading indicator of meat proterties, be related to animal in producing such as numerous economic characters such as lean ratio, the speed of growth, feed conversion rate, intramuscular fats.In the long-term production practice, people find on the different animal product price of lipid content very big difference is arranged already.People's consumption habit has been transferred to from happiness food fat meat and has been liked to eat lean meat, makes the economic benefit of fat individuality in the process that animal produces descend like this.Fat has become a kind of byproduct in producing for animal.It has reduced feed conversion rate, has reduced the economic benefit of animal production process simultaneously.Made significant headway in the fatty character breeding process in the past few decades.Along with the develop rapidly of molecular biology theory and technology, provide another method to improve the hereditary basis of animal to us.(obese, ob), (Melanocortin-4 receptor MC4R) has certain influence to fatty character to melanocyte cortical hormone receptor 4 genes of finding in 1998 to the ob gene of nineteen fifty discovery.
Excessive Intake cholesterol and heart trouble are contacted directly, and are to cause arteriosclerosis and cardiopathic main reason.Cardiovascular disordeies such as heart trouble, arteriosclerosis and apoplexy cause dead number annual nearly 1,000,000, have reached 2500 people average every day! And cholesterol level is very high in the animal food, eats the generation that too much animal food can cause above-mentioned disease.Egg healthy, low-cholesterol comes out, but how to produce the also not further research of pork of low-cholesterol.Undoubtedly, healthy, low-cholesterol pork all can bring huge economic benefit and social benefit.Directed and differentiation factor 1 (the Adipocyte Detetmination andDifferentiation factor-1 of adipocyte, ADD1) gene claims sterol regulatory element conjugated protein 1 (SterolRegulatory Element Binding proteins-1 SREBP1) gene again, directed and the differentiation factor 1 of coding adipocyte, ADD1 is a kind of important nuclear factor in the adipocyte atomization, strengthens the atomization that conjugated protein (C/EBPs) controls adipocyte jointly with peroxisome proliferation albumen activated receptor (PPAR γ) and CCAAT element.In addition, ADD1 can also regulate the intravital cholesterol levels of animal by the sterol regulatory element.
In analysis to the prior art document, though " Comparative Biochemistry and PhysiologyPart B; 1999; 28:307-318 " affirms fully the differentiation of DD1 gene pairs adipocyte and the function of directive action, particularly the ADD1 gene pairs regulate other and fatty deposits the genes involved function determine make the application of people aspect fatty deposits obtain rapid progress.But not finding as yet so far has polymorphism in the ADD1 gene order, can not carry out the genotype in mutational site and linkage analysis that proterties is said and carry out marker assisted selection.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, the gene order of a kind of porcine adipocyte orientation and differentiation factor 1 is provided.The adipocyte orientation of pig disclosed in this invention and cDNA sequence, the aminoacid sequence of differentiation factor 1, the regulating effect of pig ADD1 gene protein encoding sequence in the adipocyte atomization made specified, make it can pass through the albumen of this gene of preparation on the one hand, be used for this gene in porcine adipocyte differentiation and control agent inner cholesterol level, the method that can pass through marker assisted selection (MAS) on the other hand acts on performance is guiding in the production of bacon hogs and low cholesterol type pig and the breeding.
The present invention is achieved by the following technical solutions, of the present invention is the ADD1 gene that amplifies from the total RNA of the spleen cell of pig, the aminoacid sequence (as follows) of its cDNA sequence and proteins encoded, this full length gene 3830bp, open reading frame 11bp-3453bp section, coding has 1151 amino acid whose protein, the long 10bp of its 3 ' end non-coding sequence, the long 366bp of 5 ' end non-coding sequence.Helix-loop-helix (Helix-Loop-Helix, core texture HLH) are arranged at the 320-380 place of protein sequence.
The search homologous sequence is 99.9% with the partial sequence homology of known two ADD1 genes in GenBank, it is GenBank AY307771 and 99.1%, be GenBank AF102873, sequence homology 82.9% with the people, be GenBank_NM 004176, homology with mouse is 72.8%, be GenBank U09103, homology with chicken is 65.4%, i.e. GenBank AY029224, AF103873 relatively, 1022 g-a at full length sequence, 1034 t-c, 108 a-g have sudden change, relative AY307771,532 a-g, 625 c-t have sudden change, and 600 c-t sudden changes cause aminoacid sequence to produce into the sudden change of Leu-Pro.Above mutational site all might be used in marker assisted selection.
The cDNA sequence of ADD1 gene is as follows:
CGCCGGCGCC?ATGGACGAGC?CGCCCTTCAC?AGAGGCAGCC?TTGGAGCAGG?CGCTGGCCGA 60
GCCATGCGAG?CTGGACGCGG?CGCTGCTGAC?CGACATCGAA?GACATGCTTC?AGCTCATCAA 120
CAACCAAGAC?AGCGACTTCC?CCGGCCTGTT?CGACGCGCCC?TACGCTGGCG?TTGCAGGAGG 180
CACGGACCCC?ACCAGTCCTG?ATGCCAGCTC?CCCAGGCAGC?CCGACGCCGC?CTCCTTCCAC 240
CATGAGCTCC?CCGCTTGAAG?GCTTCCTGGG?GGGGGCCAGG?ACGCCGCCAC?CGCCCCCCGT 300
GTCCCCAACC?CAGCCTGCAC?CCACCCCGCT?GAAGATGTAC?CCGTCGGTGC?CTGCCTTCTC 360
GCCTGGGCCG?GGCATCAAGG?AGGAGCCAGT?GCCACTGACC?ATCCTGCAGC?CCCCCACCCC 420
ACAGCCCCTG?TCTGGGGCTC?TCCTGCCCCA?GAGCCTTCCA?GCCCTCGCCC?CTCCGCAGTT 480
GAGCCCTGCC?CCCGTGTTGG?GCTACCCCAG?CCCTCCTGGA?AGCTTCTCCT?CGGCGACCCC 540
TCCAGGGAGC?ACCTCACAGA?CGCTGCCTGG?CCTGCCGCTG?GCCTCTCTGC?CCGGGGTCCT 600
GCCCGTCTCC?GTGCACACCC?AGGTTCAAAG?TGCGGCCCCC?CAGCAGCTGT?TGACAGCCAC 660
GGCCACCCCC?GTGGTGTCCC?CTGGAACAAC?CACTGTGACC?TCGCAGATCC?AGCAGGTCCC 720
GGTCCTGCTG?CAGCCCCACT?TCATCAAGGC?GGACTCCCTG?CTCCTGACGA?CCATGAAAAC 780
AGACATGGGG?ACCCCTGTGA?AGGCGGCGGG?CATCGGCTCC?CTGGCCCCTG?GCACAGCCGT 840
GCAGGCAGCA?CCTTTGCAGA?CCCTGGTGAG?CGGCGGGACC?ATCCTGGCGA?CGGTGCCTCT 900
GGTAGTGGAC?ACTGACAAGC?TGCCCATCAA?CCGCCTGGCA?GCTGGGGGCA?AAGCCCTGAG 960
CTCAGGCCAG?AGCCGTGGTG?AGAAGCGGAC?GGCTCACAAT?GCCATCGAGA?AACGCTACCG 1020
CTCCTCCATC?AATGACAAGA?TCATCGAGCT?CAAGGACCTG?GTGGTGGGCA?CGGAGGCGAA 1080
GCTGAATAAA?TCCGCCGTCT?TGCGCAAGGC?CATCGACTAC?ATCCGCTTCC?TTCAGCAGAG 1140
CAACCAGAAG?CTCAAGCAGG?AGAACCTGAG?TCTGCGGACT?GCTGCCCACA?AAAGCAAGTC 1200
TCTGAAGGAC?CTGGTGTCCT?GCAGCAGCGG?AGGCCGCACC?GACGTGCCCA?TGGAGGGTGT 1260
GAAGCCGGAG?GTGGTAGACA?CCCTTAGCCC?GCCCCCCTCC?GACGCCGGCT?CGCCCTCCCA 1320
GAGCAGCCCC?TTGTCCCTCG?GCAGCAGGGG?CAGCAGCAGC?GGTGGCAGTG?GCAGTGACTC 1380
AGAGCCCGAC?AGCCCGGTTT?TTGAGGACAG?CCAGATGAAG?CCAGAGCAGC?TGCCGGCCCC 1440
CCACGGCCGG?GGCATGCTGG?ACCGCTCCCG?CCTGGCCCTG?TGCGTGCTCG?TCTTCCTCTG 1500
TCTCTCCTGC?AACCCCTTGG?CCTCACTGAT?GGGCAGCTGG?GCTCTCCCTG?GCCCCTCCGA 1560
TGCCACCAGC?GCCTACCACG?GGCCCTGGCG?CAGCGTACTG?GGCGCTGAGG?GCAGAGATGG 1620
CCCTGGCTGG?GTCCTGTGGC?TGCTGCCCCC?GCTGGTCTGG?CTGACGAATG?GGCTGCTGGT 1680
GCTGCTCTTC?TTGGCGCTTC?TCTTTGTCTA?TGGAGAGCCG?GTCACGCGGC?CCCACTCGGA 1740
TCCAGCTGTG?CGCTTCTGGA?GGCATCGCAA?GCAGGCCGAC?CTGGACCTGG?CCCGGGGGGA 1800
CTTCGCCCAG?GCTGCCCAGC?AGCTGTGGCT?GGCCCTGCGG?GCGCTGGGCC?GGCCCCTGCC 1860
CACCTCCCAC?CTGGACCTGG?CCTGCAGCCT?GCTCTGGAAC?CTCATCCGCC?ACCTGCTGCA 1920
GCGTCTCTGG?GTGGGCCGCT?GGCTGGCGGG?CCGGGCCGGG?GGCCTGCGGA?GGGACAGCGC 1980
CCTGGAGGCG?GACACCCGCA?CCAGCGCCTG?TGACGCGGCC?CTCGTCTACC?ACAAGCTGCA 2040
CCAGCTGCAC?ACCATGGGGA?AGTACCCAGG?CGGCACCTTC?GATGCCGCCA?ACCTGGCGCT 2100
GAGTGCCCTG?AACCTGGCGG?AGTGTGCGGG?TGATGCCCTG?TCCGTGGCCG?TGCTGGCGGA 2160
GGTCTATGTG?GCCGCTGCCC?TGCGAGTCAA?GACCAGTCTC?CCCCGGGCCT?TGCACTTTCT 2220
GACCCGCTTC?TTCCTGAGTA?GCGCCCGCCA?GGCCTGCCTG?GCACAAAGCG?GCTCGGTGCC 2280
TGTTGCCATG?CAGTGGCTCT?GCCACCCTGT?GGGCCAGCGT?TTCTTCGTGG?ATGGGGACTG 2340
GGCCGTGTGC?GGTGCCCCCC?GGGACAGCTT?GTACAGCGTG?GCTGGGAACC?CAGTGGACCC 2400
CCTGGCCCAG?GTGACTCAGT?TGTTCCGTGA?ACACCTCTTG?GAGCAAGCCC?TGCATTGCGT 2460
GGCCCAGCCC?AGCCCCGGCC?CTGGATCAGC?TGATGGGGAC?AGGGAATTCT?CAGAQQCCCT 2420
CGGGTTCCTG?CAGCTGCTCA?ACAGCTGTTG?TGACACAGCC?GGGGCTCCTG?CCTGCAGCTT 2580
CTCCATTGCC?TCCAGCACAG?CCGCCACGGC?CGGCACAGAC?CCGGTCGCCA?AGTGGTGGGC 2640
CTCGCTGACG?GCTGTGGTGA?CCCACTGGCT?GGGGCGGGAC?GAGGAGGCGG?CCGAGCGGCT 2700
GTACCCGCTG?GTGGAGCACC?TGCCCCGCGC?CCTGCAGGAG?TCCGAGAGAC?CCCTGCCCAG 2760
GGCAGCTCTG?CACTCCTTCA?AGGCCGCCCG?GGCCCTGCTG?GGCCGCGGGA?AGGCAGAGTC 2820
TGGCTCAGCC?AGCCTGGCCA?TGTGTGAGAA?GGCCAGTGGG?TACCTGCAGG?ACAGCCTGGC 2880
CGCCACGCCA?GCCGGCAGCT?CCATTGACAA?GGCCATGCAG?CTGCTCCTGT?GTGTCCTGCT 2940
CCTTGTGGCG?CGCACCAGCC?TGTGGCAGCG?GCAGAAGCCG?CCCCCACCCT?CCCAGGCCTC 3000
GCAGGGCTCG?AGCAGCGGGG?CCCAGGCCTC?TGCGCTCGAG?CTGCGCGGCT?TCCAGAGGGA 3060
CCTGAGTGGC?CTGAGGCGCC?TGGCGCAGAG?CTTCCGGCCT?GCCATGCGGA?GGGTGTTCCT 3120
CCATGAGGCC?ACCGCCAGGC?TGATGGCCGG?GGCCAGCCCC?GCGCGGACGC?ACCAGCTCCT 3180
GGACCGCAGT?CTGAGGAGGA?GGGCGGGCCC?CTGCGGCAGA?GGAGGCGCGG?CGGCCGCGGC 3240
GGCGGCGGAG?CTGGAACCGC?GGCCCACGCG?GCGGGAGCAG?GCCGAGGCCC?TGCTGCTCGC 3300
CTCCTGCTAC?CTGCCGCCCG?GCTTCCTGTC?GGCGCCGGGG?CAGCGCGTGG?GCATGCTGGC 3360
CGAGGCGGCG?CGCACGCTCG?AGAAGCTCGG?CGACCGCCGG?CTGCTGCACG?ACTGCCAGCA 3420
GATGCTCATG?CGCCTGGGCG?GCGGGACCAC?CGTGACCTCC?AGCTAGACCC?CTGCGGGCGG 3480
CCCAGCCCCA?GCGACCCCGT?CTCAGTCTCG?ACGGCCGAGG?GCAGCTCTGG?AGCCCCGGCC 3540
CCCGGCTCGC?CGGCGGAGTG?CCCGGTCCCT?GGGGGTATGG?AGACGCGGGG?CCCGGCGTTG 3600
ACGGGGCCGT?CCAGCCCGCA?CCCCGGGTCG?ACGAGCGGCC?GCCAGGGTGG?TGCTGGCCTC 3660
TGGTGGCCGG?CCGAGGGATT?GCCGGGGCCT?GGCGGCCGCC?AGGTGCCTCC?CATTCCACCC 3720
ACTGGCAGCC?ACTCCGCGGC?TTTCCCTCTC?TGTCCAGGGG?GAGGAAAGGG?GTGCGTTTCA 3780
CTGGGCCGAG?GGGCCGGCTT?GGCCTTCCTT?GCCGCCGGGC?GCCTCCTTCC 3830
The aminoacid sequence of ADD1 gene coded protein is as follows:
MDEPPFTEAA?LEQALAEPCE?LDAALLTDIE?DMLQLINNQD?SDFPGLFDAP?YAGVAGGTDP 60
TSPDASSPGS?PTPPPSTMSS?PLEGFLGGAR?TPPPPPVSPT?QPAPTPLKMY?PSVPAFSPGP 120
GIKEEPVPLT?ILQPPTPQPL?SGALLPQSLP?ALAPPQLSPA?PVLGYPSPPG?SFSSATPPGS 180
TSQTLPGLPL?ASLPGVLPVS?VHTQVQSAAP?QQLLTATATP?VVSPGTTTVT?SQIQQVPVLL 240
QPHFIKADSL?LLTTMKTDMG?TPVKAAGIGS?LAPGTAVQAA?PLQTLVSGGT?ILATVPLVVD 300
TDKLPINRLA?AGGKALSSGQ?SRGEKRTAHN?AIEKRYRSSI?NDKIIELKDL?VVGTEAKLNK 360
SAVLRKAIDY?IRFLQQSNQK?LKQENLSLRT?AAHKSKSLKD?LVSCSSGGRT?DVPMEGVKPE 420
VVDTLSPPPS?DAGSPSQSSP?LSLGSRGSSS?GGSGSDSEPD?SPVFEDSQMK?PEQLPAPHGR 480
GMLDRSRLAL?CVLVFLCLSC?NPLASLMGSW?ALPGPSDATS?AYHGPWRSVL?GAEGRDGPGW 540
VLWLLPPLVW?LTNGLLVLLF?LALLFVYGEP?VTRPHSDPAV?RFWRHRKQAD?LDLARGDFAQ 600
AAQQLWLALR?ALGRPLPTSH?LDLACSLLWN?LIRHLLQRLW?VGRWLAGRAG?GLRRDSALEA 660
DTRTSACDAA?LVYHKLHQLH?TMGKYPGGHL?DAANLALSAL?NLAECAGDAL?SVAVLAEVYV 720
AAALRVKTSL?PRALHFLTRF?FLSSARQACL?AQSGSVPVAM?QWLCHPVGHR?FFVDGDWAVC 780
GAPRDSLYSV?AGNPVDPLAQ?VTQLFREHLL?EQALHCVAQP?SPGPGSADGD?REFSEALGFL 840
QLLNSCCDTA?GAPACSFSIA?SSTAATAGTD?PVAKWWASLT?AVVTHWLGRD?EEAAERLYPL 900
VEHLPRALQE?SERPLPRAAL?HSFKAARALL?GRGKAESGSA?SLAMCEKASG?YLQDSLAATP 960
AGSSIDKAMQ?LLLCDLLLVA?RTSLWQRQKP?PPPSQASQGS?SSGAQASALE?LRGFQRDLSG 1020
LRRLAQSFRP?AMRRVFLHEA?TARLMAGASP?ARTHQLLDRS?LRRRAGPCGR?GGAAAAAAAE 1080
LEPRPTRREQ?AEALLLASCY?LPPGFLSAPG?QRVGMLAEAA?RTLEKLGDRR?LLHDCQQMLM 1140
RLGGGTTVTS?S 1151
The present invention has substantive distinguishing features and marked improvement, at present, has only the cDNA sequence (GenBank AY307771 and AF102873) of part in the research of pig ADD1 gene.Invention clones full length cDNA sequence first.An aspect of of the present present invention can be used for studying the effect of this gene in processes such as porcine adipocyte differentiation and control agent inner cholesterol levels by the albumen of this gene of preparation.The method that can pass through marker assisted selection (MAS) on the other hand is to performance directiveness effect in the production of bacon hogs and low cholesterol type pig and the breeding.
Description of drawings
Fig. 1 is pig spleen total tissue RNA agarose electrophoresis figure
The ratio of 28S and 18S RNA brightness is approximately 2 among the figure, illustrates that total tissue RNA is not degraded, and can carry out reverse transcription.
Fig. 2 is the agarose electrophoresis figure that template is carried out the PCR reaction product with the reverse transcription of the total RNA of spleen tissue
Through the gel imaging system analysis, the segment size is 750bp.
Fig. 3 pig is with the cluster analysis figure between encoding sequence between the ADD1 gene (SREBF1 gene) of other species and SREBF2 gene.
Fig. 4 is for to obtain helix-loop-helix (Helix-Loop-Helix, HLH) three-dimensional plot of core texture by the rpsBlast software analysis.
Embodiment
Below in conjunction with specific embodiment the present invention is further described: these embodiment only are used to the present invention is described and are not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, for example the Sambrook equimolecular is cloned: laboratory manual (New York:Cold Spring Harbor LaboratoryPress, 1989) condition described in, or the condition of advising according to manufacturer.
Embodiment 1
The clone of pig ADD1 gene
1. separate tissue (Isolation)
Gather the fresh spleen tissue (kind is " a plum mountain pig ") of pig from the slaughterhouse and put into the freezing preservation of liquid nitrogen rapidly, and it is taken back the laboratory, put into-70 ℃ of preservations, prepare to extract total tissue RNA.
2. the separation (Total RNA isolation) of total RNA
(1) preparation work of extraction RNA
The NaOH soaked overnight of used in glass products 0.1M is washed repeatedly with tap water, uses distilled water flushing again 2 times, and 180 ℃ were toasted 4 hours.
Homogenizer, electrophoresis chamber be with 3% hydrogen peroxide dipping 20-30 minute, again with 0.1% DEPC water flushing.Because the DEPC of deactivation is not influential to the PCR reaction, available 0.5% SDS handles.
The DEPC water logging bubble of Tip head, EP effective 0.1% spends the night, and autoclaving makes the DEPC inactivation.
Distilled water and solution are handled with DEPC, and promptly every 100ml water or solution add 0.1mlDEPC liquid, and room temperature is placed and spent the night, and autoclaving made the DEPC inactivation in 30 minutes.
(2) total tissue RNA is extracted
Getting 100mg organizes the sample adding that 1ml Trizol (trizal reagent, invitrogen BRL, USA) homogenate in homogenizer are arranged at liquid nitrogen cryopreservation.4 ℃ 12000 rev/mins centrifugal 10 minutes.Draw supernatant liquor, 15-30 ℃ of incubation 5 minutes.Add the 0.2ml chloroform, cover lid is used hand concuss 15 seconds.15-30 ℃ incubation 2-3 minute.4 ℃ 12000 rev/mins centrifugal 15 minutes.Draw supernatant liquor, add the 0.8ml Virahol, mix.15-30 ℃ of incubation 10 minutes, 4 ℃ 12000 rev/mins centrifugal 10 minutes, centrifuge tube bottom white precipitate is RNA.Outwell supernatant, add 1ml 75% washing with alcohol RNA, 4 ℃ 7500 rev/mins 10 minutes.Packing among seasoning or the vacuum-drying RNA. water-soluble (RNase free) ,-80 ℃ of preservations.
(3) evaluation of total tissue RNA
By measuring rna content on the spectrophotometer and with the quality of agarose electrophoretic analysis RNA, concrete grammar is as follows:
Electrophoresis chamber soaked 4-5 hour with RNA scavenging solution (100mM NaOH, 1mM EDTA), washed repeatedly with the DEPC treated water that to pour 1 * TBE into behind the several stand-by again.Sepharose is prepared with 1 * TBE.The 10ul sample-loading buffer (2 * TBE, 13% phenanthrene can, 0.1% bromjophenol blue, 7M urea) in add the above total tissue RNA of 1ul, 65 ℃ of sex change were put into after 10 minutes frozen water 2-3 minute immediately.Point sample is electrophoresis in sepharose.
3. Full Length cDNA Cloning (Cloning of Full-length cDNA)
According to cDNA (GenBank AY307771) the design reverse transcription primer of the known portions of pig (5 '-ACCGTCGCCAGGAT-3 ') and PCR downstream primer (5 '-CAGGGAGTCCGCCGTTGAT-3 '), utilize homologous genes clone principle according to the amino acid conserved sequence of people and mouse design PCR upstream primer (5 '-CGCCGGCGCCATGGACGAGC-3 '), adopt RT-PCR to carry out the cDNA full-length clone, divide three phases to carry out with the method that 5 '-RACE (TaKaRa test kit) combines:
(1) cDNA's is synthetic
Utilize the total RNA 5ug of spleen tissue to dispose reaction solution and carry out the PCR reaction according to TaKaRa 5 '-RACE test kit.
(2) decomposition of heterozygosis chain RAN
Utilize the first step reaction solution according to TaKaRa 5 '-RACE test kit configuration reaction solution, with 30 ℃ of digestion of Rnase H 1 hour, behind the RNA in the digestion heterozygosis chain, with the single stranded DNA diluted for use.
(3) PCR reaction
Reaction solution with second step of diluting is that template is carried out the PCR reaction.PCR[BP001(SEQ?ID?NO.1)+BP002(SEQ?ID?NO.2)]。Its reaction parameter is: 95 ℃ of pre-sex change 5min, 95 ℃ of sex change 30s, 56 ℃ of annealing 30s, 72 ℃ are extended 1min, 35 circulations, and 72 ℃ are extended 10min, 4 ℃ of 10min. obtain the dna segment of 760bp, wherein have 276bp with the known segment homology of pig up to 98.91%
(4) order-checking
The PCR product is reclaimed the back be connected, be transformed in the competent e. coli jm109 with PMD 18-T Vector (TaKaRa).In 37 ℃ of overnight incubation of LB substratum, deliver order-checking through the bacterial strain after bacterial strain PCR and the double digestion evaluation.Sequencing result is total to 760bp through the splicing back.
Embodiment 2
Pig ADD1 Gene Sequence Analysis and homology type are relatively
1, pig ADD1 Gene Sequence Analysis
On http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi, carry out structure prediction after CDS in the cDNA sequence partly become aminoacid sequence with primer priemer 5.0 software translations with rpsBlast software.Discovery has helix-loop-helix (Helix-Loop-Helix, core texture HLH) (seeing accompanying drawing 4) at the 320-380 place of aminoacid sequence.
2, the homology analysis of pig ADD1 gene order
This aminopeptidase gene acid sequence and base sequence are carried out homology analysis with Blastn and Blastp software respectively on http://www.ncbi.nlm.nih.gov/BLAST/BLAST/Blast.cgi.Found that the base sequence of this gene and aminoacid sequence have high homology with the ADD1 gene of people and mouse.
3, the structure of phylogenetic tree
Come drawing system to set the ADD1 gene and SREBP2 gene (another one gene in the SREBPs family) sequence of different plant species with DNAMAN software.Found that homology is higher between pig ADD1 gene is with Mammals people, dog and musculus cdna, and have only 66% with the homology of chicken.Homology has 51% between two genes in the SREBPs gene family.(seeing accompanying drawing 3) result shows that the resulting sequence of experiment is a target sequence.
Finding mutational site, 6 place with the homology of the sequence of original pig in relatively.Find with AF103873 is relatively middle, in 1022 g-a of full length sequence, the sudden change of 1034 t-c, 108 a-g.With the relatively middle sudden change of finding 532 a-g, 625 c-t of AY307771,600 c-t sudden changes cause the sudden change of Leu-Pro.Above mutational site all might be used in marker assisted selection.
Table 1
Same AF103873
Query: 994?tcacaatgccatcgagaaacgctaccgctcctccatcaatgacaagatcatcgagctcaa?1053
||||||||||||||||||?|||||||||||?|||||||||||||||||||||||||||||
AF103873:301?tcacaatgccatcgagaagcgctaccgctcttccatcaatgacaagatcatcgagctcaa?360
Query:1054?ggacctggtggtgggcacggaggcgaagctgaataaatccgccgtcttgcgcaaggccat?1113
|||||||||||||||||||||||||||?|||||||||||||||||||||||||||||||
AF103873:361?ggacctggtggtgggcacggaggcgaaactgaataaatccgccgtcttgcgcaaggccat?420
Same AY307771
Query: 485?cctgcccccgtgttgggctaccccagccctcctggaagcttctcctcggcgacccctcca?544
|||||||||||||||||||||||||||||||||||||||||||||||?||||||||||||
AY307771 1 cctgcccccgtgttgggctaccccagccctcctggaagcttctcctcagcgacccctcca?60
Query: 545?gggagcacctcacagacgctgcctggcctgccgctggcctctctgcccggggtcctgccc?604
|||||||||||||||||||||||||||||||||||||||||||||||||||||||?||||
AY307771:61 gggagcacctcacagacgctgcctggcctgccgctggcctctctgcccggggtcccgccc?120
Query: 605?gtctccgtgcacacccaggttcaaagtgcggccccccagcagctgttgacagccacggcc?664
||||||||||||||||||||?|||||||||||||||||||||||||||||||||||||||
AY307771:121?gtctccgtgcacacccaggtccaaagtgcggccccccagcagctgttgacagccacggcc?180
Query: the nucleotide sequence of the porcine adipocyte differentiation and the directed factor 1
Sequence that the present invention relates to and mark apportion are as follows:
(1) information of SEQ ID NO.1
(i) sequence signature:
(A) length: 14bp
(B) type: Nucleotide
(C) chain: strand
(D) topological framework: linearity
(ii). molecule type: oligonucleotide
(iii). sequence description: SEQ ID NO.1
ACCGTCGCCAGGAT
(2) information of SEQ ID NO.2
(i) sequence signature:
(A) length: 20bp
(B) type: Nucleotide
(C) chain: strand
(D) topological framework: linearity
(ii). molecule type: oligonucleotide
(iii). sequence description: SEQ ID NO.2
CGCCGGCGCCATGGACGAGC
(3) information of SEQ ID NO.3
(i) sequence signature:
(A) length: 18bp
(B) type: Nucleotide
(C) chain: strand
(D) topological framework: linearity
(ii). molecule type: oligonucleotide
(iii). sequence description: SEQ ID NO.3
CAGGGAGTCCGCCTTGAT
(4) information of SEQ ID NO.4
<110〉Shanghai Communications University
<120〉pig ADD1 gene protein encoding sequence
<140>
<141>2004-01-6
<160>3
<170>PatentIn?version?3.1
<210>1
<211>3830
<212>DNA
<213〉pig Sus scrofa ADD1 albumen
<400>1
CGCCGGCGCC?ATGGACGAGC?CGCCCTTCAC?AGAGGCAGCC?TTGGAGCAGG?CGCTGGCCGA 60
GCCATGCGAG?CTGGACGCGG?CGCTGCTGAC?CGACATCGAA?GACATGCTTC?AGCTCATCAA 120
CAACCAAGAC?AGCGACTTCC?CCGGCCTGTT?CGACGCGCCC?TACGCTGGCG?TTGCAGGAGG 180
CACGGACCCC?ACCAGTCCTG?ATGCCAGCTC?CCCAGGCAGC?CCGACGCCGC?CTCCTTCCAC 240
CATGAGCTCC?CCGCTTGAAG?GCTTCCTGGG?GGGGGCCAGG?ACGCCGCCAC?CGCCCCCCGT 300
GTCCCCAACC?CAGCCTGCAC?CCACCCCGCT?GAAGATGTAC?CCGTCGGTGC?CTGCCTTCTC 360
GCCTGGGCCG?GGCATCAAGG?AGGAGCCAGT?GCCACTGACC?ATCCTGCAGC?CCCCCACCCC 420
ACAGCCCCTG?TCTGGGGCTC?TCCTGCCCCA?GAGCCTTCCA?GCCCTCGCCC?CTCCGCAGTT 480
GAGCCCTGCC?CCCGTGTTGG?GCTACCCCAG?CCCTCCTGGA?AGCTTCTCCT?CGGCGACCCC 540
TCCAGGGAGC?ACCTCACAGA?CGCTGCCTGG?CCTGCCGCTG?GCCTCTCTGC?CCGGGGTCCT 600
GCCCGTCTCC?GTGCACACCC?AGGTTCAAAG?TGCGGCCCCC?CAGCAGCTGT?TGACAGCCAC 660
GGCCACCCCC?GTGGTGTCCC?CTGGAACAAC?CACTGTGACC?TCGCAGATCC?AGCAGGTCCC 720
GGTCCTGCTG?CAGCCCCACT?TCATCAAGGC?GGACTCCCTG?CTCCTGACGA?CCATGAAAAC 780
AGACATGGGG?ACCCCTGTGA?AGGCGGCGGG?CATCGGCTCC?CTGGCCCCTG?GCACAGCCGT 840
GCAGGCAGCA?CCTTTGCAGA?CCCTGGTGAG?CGGCGGGACC?ATCCTGGCGA?CGGTGCCTCT 900
GGTAGTGGAC?ACTGACAAGC?TGCCCATCAA?CCGCCTGGCA?GCTGGGGGCA?AAGCCCTGAG 960
CTCAGGCCAG?AGCCGTGGTG?AGAAGCGGAC?GGCTCACAAT?GCCATCGAGA?AACGCTACCG 1020
CTCCTCCATC?AATGACAAGA?TCATCGAGCT?CAAGGACCTG?GTGGTGGGCA?CGGAGGCGAA 1080
GCTGAATAAA?TCCGCCGTCT?TGCGCAAGGC?CATCGACTAC?ATCCGCTTCC?TTCAGCAGAG 1140
CAACCAGAAG?CTCAAGCAGG?AGAACCTGAG?TCTGCGGACT?GCTGCCCACA?AAAGCAAGTC 1200
TCTGAAGGAC?CTGGTGTCCT?GCAGCAGCGG?AGGCCGCACC?GACGTGCCCA?TGGAGGGTGT 1260
GAAGCCGGAG?GTGGTAGACA?CCCTTAGCCC?GCCCCCCTCC?GACGCCGGCT?CGCCCTCCCA 1320
GAGCAGCCCC?TTGTCCCTCG?GCAGCAGGGG?CAGCAGCAGC?GGTGGCAGTG?GCAGTGACTC 1380
AGAGCCCGAC?AGCCCGGTTT?TTGAGGACAG?CCAGATGAAG?CCAGAGCAGC?TGCCGGCCCC 1440
CCACGGCCGG?GGCATGCTGG?ACCGCTCCCG?CCTGGCCCTG?TGCGTGCTCG?TCTTCCTCTG 1500
TCTCTCCTGC?AACCCCTTGG?CCTCACTGAT?GGGCAGCTGG?GCTCTCCCTG?GCCCCTCCGA 1560
TGCCACCAGC?GCCTACCACG?GGCCCTGGCG?CAGCGTACTG?GGCGCTGAGG?GCAGAGATGG 1620
CCCTGGCTGG?GTCCTGTGGC?TGCTGCCCCC?GCTGGTCTGG?CTGACGAATG?GGCTGCTGGT 1680
GCTGCTCTTC?TTGGCGCTTC?TCTTTGTCTA?TGGAGAGCCG?GTCACGCGGC?CCCACTCGGA 1740
TCCAGCTGTG?CGCTTCTGGA?GGCATCGCAA?GCAGGCCGAC?CTGGACCTGG?CCCGGGGGGA 1800
CTTCGCCCAG?GCTGCCCAGC?AGCTGTGGCT?GGCCCTGCGG?GCGCTGGGCC?GGCCCCTGCC 1860
CACCTCCCAC?CTGGACCTGG?CCTGCAGCCT?GCTCTGGAAC?CTCATCCGCC?ACCTGCTGCA 1920
GCGTCTCTGG?GTGGGCCGCT?GGCTGGCGGG?CCGGGCCGGG?GGCCTGCGGA?GGGACAGCGC 1980
CCTGGAGGCG?GACACCCGCA?CCAGCGCCTG?TGACGCGGCC?CTGCTCTACC?ACAAGCTGCA 2040
CCAGCTGCAC?ACCATGGGGA?AGTACCCAGG?CGGGCACCTC?GATGCCGCCA?ACCTGGCGCT 2100
GAGTGCCCTG?AACCTGGCGG?AGTGTGCGGG?TGATGCCCTG?TCCGTGGCCG?TGCTGGCGGA 2160
GGTCTATGTG?GCCGCTGCCC?TGCGAGTCAA?GACCAGTCTC?CCCCGGGCCT?TGCACTTTCT 2220
GACCCGCTTC?TTCCTGAGTA?GCGCCCGCCA?GGCCTGCCTG?GCACAAAGCG?GCTCGGTGCC 2280
TGTTGCCATG?CAGTGGCTCT?GCCACCCTGT?GGGCCACCGT?TTCTTCGTGG?ATGGGGACTG 2340
GGCCGTGTGC?GGTGCCCCCC?GGGACAGCTT?GTACAGCGTG?GCTGGGAACC?CAGTGGACCC 2400
CCTGGCCCAG?GTGACTCAGT?TGTTCCGTGA?ACACCTCTTG?GAGCAAGCCC?TGCATTGCGT 2460
GGCCCAGCCC?AGCCCCGGCC?CTGGATCAGC?TGATGGGGAC?AGGGAATTCT?CAGAGGCCCT 2420
CGGGTTCCTG?CAGCTGCTCA?ACAGCTGTTG?TGACACAGCC?GGGGCTCCTG?CCTGCAGCTT 2580
CTCCATTGCC?TCCAGCACAG?CCGCCACGGC?CGGCACAGAC?CCGGTCGCCA?AGTGGTGGGC 2640
CTCGCTGACG?GCTGTGGTGA?CCCACTGGCT?GGGGCGGGAC?GAGGAGGCGG?CCGAGCGGCT 2700
GTACCCGCTG?GTGGAGCACC?TGCCCCGCGC?CCTGCAGGAG?TCCGAGAGAC?CCCTGCCCAG 2760
GGCAGCTCTG?CACTCCTTCA?AGGCCGCCCG?GGCCCTGCTG?GGCCGCGGGA?AGGCAGAGTC 2820
TGGCTCAGCC?AGCCTGGCCA?TGTGTGAGAA?GGCCAGTGGG?TACCTGCAGG?ACAGCCTGGC 2880
CGCCACGCCA?GCCGGCAGCT?CCATTGACAA?GGCCATGCAG?CTGCTCCTGT?GTGACCTGCT 2940
CCTTGTGGCG?CGCACCAGCC?TGTGGCAGCG?GCAGAAGCCG?CCCCCACCCT?CCCAGGCCTC 3000
GCAGGGCTCG?AGCAGCGGGG?CCCAGGCCTC?TGCGCTCGAG?CTGCGCGGCT?TCCAGAGGGA 3060
CCTGAGTGGC?CTGAGGCGCC?TGGCGCAGAG?CTTCCGGCCT?GCCATGCGGA?GGGTGTTCCT 3120
CCATGAGGCC?ACCGCCAGGC?TGATGGCCGG?GGCCAGCCCC?GCGCGGACGC?ACCAGCTCCT 3180
GGACCGCAGT?CTGAGGAGGA?GGGCGGGCCC?CTGCGGCAGA?GGAGGCGCGG?CGGCCGCGGC 3240
GGCGGCGGAG?CTGGAACCGC?GGCCCACGCG?GCGGGAGCAG?GCCGAGGCCC?TGCTGCTCGC 3300
CTCCTGCTAC?CTGCCGCCCG?GCTTCCTGTC?GGCGCCGGGG?CAGCGCGTGG?GCATGCTGGC 3360
CGAGGCGGCG?CGCACGCTCG?AGAAGCTCGG?CGACCGCCGG?CTGCTGCACG?ACTGCCAGCA 3420
GATGCTCATG?CGCCTGGGCG?GCGGGACCAC?CGTGACCTCC?AGCTAGACCC?CTGCGGGCGG 3480
CCCAGCCCCA?GCGACCCCGT?CTCAGTCTCG?ACGGCCGAGG?GCAGCTCTGG?AGCCCCGGCC 3540
CCCGGCTCGC?CGGCGGAGTG?CCCGGTCCCT?GGGGGTATGG?AGACGCGGGG?CCCGGCGTTG 3600
ACGGGGCCGT?CCAGCCCGCA?CCCCGGGTCG?ACGAGCGGCC?GCCAGGGTGG?TGCTGGCCTC 3660
TGGTGGCCGG?CCGAGGGATT?GCCGGGGCCT?GGCGGCCGCC?AGGTGCCTCC?CATTCCACCC 3720
ACTGGCAGCC?ACTCCGCGGC?TTTCCCTCTC?TGTCCAGGGG?GAGGAAAGGG?GTGCGTTTCA 3780
CTGGGCCGAG?GGGCCGGCTT?GGCCTTCCTT?GCCGCCGGGC?GCCTCCTTCC 3830
<210>2
<211>1151
<212>PRT
<213〉pig Sus scrofa ADD1 albumen
<400>2
MDEPPFTEAA?LEQALAEPCE?LDAALLTDIE?DMLQLINNQD?SDFPGLFDAP?YAGVAGGTDP 60
TSPDASSPGS?PTPPPSTMSS?PLEGFLGGAR?TPPPPPVSPT?QPAPTPLKMY?PSVPAFSPGP 120
GIKEEPVPLT?ILQPPTPQPL?SGALLPQSLP?ALAPPQLSPA?PVLGYPSPPG?SFSSATPPGS 180
TSQTLPGLPL?ASLPGVLPVS?VHTQVQSAAP?QQLLTATATP?VVSPGTTTVT?SQIQQVPVLL 240
QPHFIKADSL?LLTTMKTDMG?TPVKAAGLGS?LAPGTAVQAA?PLQTLVSGGT?ILATVPLVVD 300
TDKLPINRLA?AGGKALSSGQ?SRGEKRTAHN?AIEKRYRSSI?NDKIIELKDL?VVGTEAKLNK 360
SAVLRKAIDY?IRFLQQSNQK?LKQENLSLRT?AAHKSKSLKD?LVSCSSGGRT?DVPMEGVKPE 420
VVDTLSPPPS?DAGSPSQSSP?LSLGSRGSSS?GGSGSDSEPD?SPVFEDSQMK?PEQLPAPHGR 480
GMLDRSRLAL?CVLVFLCLSC?NPLASLMGSW?ALPGPSDATS?AYHGPWRSVL?GAEGRDGPGW 540
VLWLLPPLVW?LTNGLLVLLF?LALLFVYGEP?VTRPHSDPAV?RFWRHRKQAD?LDLARGDFAQ 600
AAQQLWLALR?ALGRPLPTSH?LDLACSLLWN?LIRHLLQRLW?VGRWLAGRAG?GLRRDSALEA 660
DTRTSACDAA?LVYHKLHQLH?TMGKYPGGHL?DAANLALSAL?NLAECAGDAL?SVAVLAEVYV 720
AAALRVKTSL?PRALHFLTRF?FLSSARQACL?AQSGSVPVAM?QWLCHPVGHR?FFVDGDWAVC 780
GAPRDSLYSV?AGNPVDPLAQ?VTQLFREHLL?EQALHCVAQP?SPGPGSADGD?REFSEALGFL 840
QLLNSCCDTA?GAPACSFSIA?SSTAATAGTD?PVAKWWVSLT?AVVTHWLGRD?EEAAERLYPL 900
VEHLPRALQE?SERPLPRAAL?HSFKAARALL?GRGKAESGSA?SLAMCEKASG?YLQDSLAATP 960
AGSSIDKAMQ?LLLCDLLLVA?RTSLWQRQKP?PPPSQASQGS?SSGAQASALE?LRGFQRDLSG 1020
LRRLAQSFRP?AMRRVFLHEA?TARLMAGASP?ARTHQLLDRS?LRRRAGPCGR?GGAAAAAAAE 1080
LEPRPTRREQ?AEALLLASCY?LPPGFLSAPG?QRVGMLAEAA?RTLEKLGDRR?LLHDCQQMLM 1140
RLGGGTTVTS?S 1151
Claims (4)
1, the gene order of a kind of porcine adipocyte orientation and differentiation factor 1, it is characterized in that, the ADD1 gene that from the total RNA of the spleen cell of pig, amplifies, the aminoacid sequence of its cDNA sequence and proteins encoded, this full length gene 3830bp, open reading frame 11bp-3453bp section, coding has 1151 amino acid whose protein, the long 10bp of its 3 ' end non-coding sequence, 5 ' holds the long 366bp of non-coding sequence, and the core texture of helix-loop-helix is arranged at the 320-380 place of protein sequence.
2, the gene order of porcine adipocyte orientation according to claim 1 and differentiation factor 1, it is characterized in that, the search homologous sequence is 99.9% with the partial sequence homology of known two ADD1 genes in GenBank, it is GenBank AY307771 and 99.1%, be GenBank AF102873, sequence homology 82.9% with the people, be GenBank_NM 004176, homology with mouse is 72.8%, be GenBank U09103, homology with chicken is 65.4%, i.e. GenBank AY029224, AF103873 relatively, 1022 g-a at full length sequence, 1034 t-c, 108 a-g have sudden change, relative AY307771,532 a-g, 625 c-t have sudden change, and 600 c-t sudden changes cause aminoacid sequence to produce into the sudden change of Leu-Pro.
3, the gene order of porcine adipocyte orientation according to claim 1 and differentiation factor 1 is characterized in that, the cDNA sequence of ADD1 gene:
CGCCGGCGCCATGGACGAGCCGCCCTTCACAGAGGCAGCCTTGGAGCAGGCGCTGGCCGAGCCATGCGAGCTGGACGCGGC
GCTGCTGACCGACATCGAAGACATGCTTCAGCTCATCAACAACCAAGACAGCGACTTCCCCGGCCTGTTCGACGCGCCCTA
CGCTGGCGTTGCAGGAGGCACGGACCCCACCAGTCCTGATGCCAGCTCCCCAGGCAGCCCGACGCCGCCTCCTTCCACCAT
GAGCTCCCCGCTTGAAGGCTTCCTGGGGGGGGCCAGGACGCCGCCACCGCCCCCCGTGTCCCCAACCCAGCCTGCACCCAC
CCCGCTGAAGATGTACCCGTCGGTGCCTGCCTTCTCGCCTGGGCCGGGCATCAAGGAGGAGCCAGTGCCACTGACCATCCT
GCAGCCCCCCACCCCACAGCCCCTGTCTGGGGCTCTCCTGCCCCAGAGCCTTCCAGCCCTCGCCCCTCCGCAGTTGAGCCC
TGCCCCCGTGTTGGGCTACCCCAGCCCTCCTGGAAGCTTCTCCTCGGCGACCCCTCCAGGGAGCACCTCACAGACGCTGCC
TGGCCTGCCGCTGGCCTCTCTGCCCGGGGTCCTGCCCGTCTCCGTGCACACCCAGGTTCAAAGTGCGGCCCCCCAGCAGCT
GTTGACAGCCACGGCCACCCCCGTGGTGTCCCCTGGAACAACCACTGTGACCTCGCAGATCCAGCAGGTCCCGGTCCTGCT
GCAGCCCCACTTCATCAAGGCGGACTCCCTGCTCCTGACGACCATGAAAACAGACATGGGGACCCCTGTGAAGGCGGCGGG
CATCGGCTCCCTGGCCCCTGGCACAGCCGTGCAGGCAGCACCTTTGCAGACCCTGGTGAGCGGCGGGACCATCCTGGCGAC
GGTGCCTCTGGTAGTGGACACTGACAAGCTGCCCATCAACCGCCTGGCAGCTGGGGGCAAAGCCCTGAGCTCAGGCCAGAG
CCGTGGTGAGAAGCGGACGGCTCACAATGCCATCGAGAAACGCTACCGCTCCTCCATCAATGACAAGATCATCGAGCTCAA
GGACCTGGTGGTGGGCACGGAGGCGAAGCTGAATAAATCCGCCGTCTTGCGCAAGGCCATCGACTACATCCGCTTCCTTCA
GCAGAGCAACCAGAAGCTCAAGCAGGAGAACCTGAGTCTGCGGACTGCTGCCCACAAAAGCAAGTCTCTGAAGGACCTGGT
GTCCTGCAGCAGCGGAGGCCGCACCGACGTGCCCATGGAGGGTGTGAAGCCGGAGGTGGTAGACACCCTTAGCCCGCCCCC
CTCCGACGCCGGCTCGCCCTCCCAGAGCAGCCCCTTGTCCCTCGGCAGCAGGGGCAGCAGCAGCGGTGGCAGTGGCAGTGA
CTCAGAGCCCGACAGCCCGGTTTTTGAGGACAGCCAGATGAAGCCAGAGCAGCTGCCGGCCCCCCACGGCCGGGGCATGCT
GGACCGCTCCCGCCTGGCCCTGTGCGTGCTCGTCTTCCTCTGTCTCTCCTGCAACCCCTTGGCCTCACTGATGGGCAGCTG
GGCTCTCCCTGGCCCCTCCGATGCCACCAGCGCCTACCACGGGCCCTGGCGCAGCGTACTGGGCGCTGAGGGCAGAGATGG
CCCTGGCTGGGTCCTGTGGCTGCTGCCCCCGCTGGTCTGGCTGACGAATGGGCTGCTGGTGCTGCTCTTCTTGGCGCTTCT
CTTTGTCTATGGAGAGCCGGTCACGCGGCCCCACTCGGATCCAGCTGTGCGCTTCTGGAGGCATCGCAAGCAGGCCGACCT
GGACCTGGCCCGGGGGGACTTCGCCCAGGCTGCCCAGCAGCTGTGGCTGGCCCTGCGGGCGCTGGGCCGGCCCCTGCCCAC
CTCCCACCTGGACCTGGCCTGCAGCCTGCTCTGGAACCTCATCCGCCACCTGCTGCAGCGTCTCTGGGTGGGCCGCTGGCT
GGCGGGCCGGGCCGGGGGCCTGCGGAGGGACAGCGCCCTGGAGGCGGACACCCGCACCAGCGCCTGTGACGCGGCCCTCGT
CTACCACAAGCTGCACCAGCTGCACACCATGGGGAAGTACCCAGGCGGGCACCTCGATGCCGCCAACCTGGCGCTGAGTGC
CCTGAACCTGGCGGAGTGTGCGGGTGATGCCCTGTCCGTGGCCGTGCTGGCGGAGGTCTATGTGGCCGCTGCCCTGCGAGT
CAAGACCAGTCTCCCCCGGGCCTTGCACTTTCTGACCCGCTTCTTCCTGAGTAGCGCCCGCCAGGCCTGCCTGGCACAAAG
CGGCTCGGTGCCTGTTGCCATGCAGTGGCTCTGCCACCCTGTGGGCCACCGTTTCTTCGTGGATGGGGACTGGGCCGTGTG
CGGTGCCCCCCGGGACAGCTTGTACAGCGTGGCTGGGAACCCAGTGGACCCCCTGGCCCAGGTGACTCAGTTGTTCCGTGA
ACACCTCTTGGAGCAAGCCCTGCATTGCGTGGCCCAGCCCAGCCCCGGCCCTGGATCAGCTGATGGGGACAGGGAATTCTC
AGAGGCCCTCGGGTTCCTGCAGCTGCTCAACAGCTGTTGTGACACAGCCGGGGCTCCTGCCTGCAGCTTCTCCATTGCCTC
CAGCACAGCCGCCACGGCCGGCACAGACCCGGTCGCCAAGTGGTGGGCCTCGCTGACGGCTGTGGTGACCCACTGGCTGGG
GCGGGACGAGGAGGCGGCCGAGCGGCTGTACCCGCTGGTGGAGCACCTGCCCCGCGCCCTGCAGGAGTCCGAGAGACCCCT
GCCCAGGGCAGCTCTGCACTCCTTCAAGGCCGCCCGGGCCCTGCTGGGCCGCGGGAAGGCAGAGTCTGGCTCAGCCAGCCT
GGCCATGTGTGAGAAGGCCAGTGGGTACCTGCAGGACAGCCTGGCCGCCACGCCAGCCGGCAGCTCCATTGACAAGGCCAT
GCAGCTGCTCCTGTGTGACCTGCTCCTTGTGGCGCGCACCAGCCTGTGGCAGCGGCAGAAGCCGCCCCCACCCTCCCAGGC
CTCGCAGGGCTCGAGCAGCGGGGCCCAGGCCTCTGCGCTCGAGCTGCGCGGCTTCCAGAGGGACCTGAGTGGCCTGAGGCG
CCTGGCGCAGAGCTTCCGGCCTGCCATGCGGAGGGTGTTCCTCCATGAGGCCACCGCCAGGCTGATGGCCGGGGCCAGCCC
CGCGCGGACGCACCAGCTCCTGGACCGCAGTCTGAGGAGGAGGGCGGGCCCCTGCGGCAGAGGAGGCGCGGCGGCCGCGGC
GGCGGCGGAGCTGGAACCGCGGCCCACGCGGCGGGAGCAGGCCGAGGCCCTGCTGCTCGCCTCCTGCTACCTGCCGCCCGG
CTTCCTGTCGGCGCCGGGGCAGCGCGTGGGCATGCTGGCCGAGGCGGCGCGCACGCTCGAGAAGCTCGGCGACCGCCGGCT
GCTGCACGACTGCCAGCAGATGCTCATGCGCCTGGGCGGCGGGACCACCGTGACCTCCAGCTAGACCCCTGCGGGCGGCCC
AGCCCCAGCGACCCCGTCTCAGTCTCGACGGCCGAGGGCAGCTCTGGAGCCCCGGCCCCCGGCTCGCCGGCGGAGTGCCCG
GTCCCTGGGGGTATGGAGACGCGGGGCCCGGCGTTGACGGGGCCGTCCAGCCCGCACCCCGGGTCGACGAGCGGCCGCCAG
GGTGGTGCTGGCCTCTGGTGGCCGGCCGAGGGATTGCCGGGGCCTGGCGGCCGCCAGGTGCCTCCCATTCCACCCACTGGC
AGCCACTCCGCGGCTTTCCCTCTCTGTCCAGGGGGAGGAAAGGGGTGCGTTTCACTGGGCCGAGGGGCCGGCTTGGCCTTC
CTTGCCGCCGGGCGCCTCCTTCC。
4, the gene order of porcine adipocyte orientation according to claim 1 and differentiation factor 1 is characterized in that, the aminoacid sequence of ADD1 gene coded protein:
MDEPPFTEAALEQALAEPCELDAALLTDIEDMLQLINNQDSDFPGLFDAPYAGVAGGTDPTSPDASSPGSPTPPPSTMSSP
LEGFLGGARTPPPPPVSPTQPAPTPLKMYPSVPAFSPGPGIKEEPVPLTILQPPTPQPLSGALLPQSLPALAPPQLSPAPV
LGYPSPPGSFSSATPPGSTSQTLPGLPLASLPGVLPVSVHTQVQSAAPQQLLTATATPVVSPGTTTVTSQIQQVPVLLQPH
FIKADSLLLTTMKTDMGTPVKAAGIGSLAPGTAVQAAPLQTLVSGGTILATVPLVVDTDKLPINRLAAGGKALSSGQSRGE
KRTAHNAIEKRYRSSINDKIIELKDLVVGTEAKLNKSAVLRKAIDYIRFLQQSNQKLKQENLSLRTAAHKSKSLKDLVSCS
SGGRTDVPMEGVKPEVVDTLSPPPSDAGSPSQSSPLSLGSRGSSSGGSGSDSEPDSPVFEDSQMKPEQLPAPHGRGMLDRS
RLALCVLVFLCLSCNPLASLMGSWALPGPSDATSAYHGPWRSVLGAEGRDGPGWVLWLLPPLVWLTNGLLVLLFLALLFVY
GEPVTRPHSDPAVRFWRHRKQADLDLARGDFAQAAQQLWLALRALGRPLPTSHLDLACSLLWNLIRHLLQRLWVGRWLAGR
AGGLRRDSALEADTRTSACDAALVYHKLHQLHTMGKYPGGHLDAANLALSALNLAECAGDALSVAVLAEVYVAAALRVKTS
LPRALHFLTRFFLSSARQACLAQSGSVPVAMQWLCHPVGHRFFVDGDWAVCGAPRDSLYSVAGNPVDPLAQVTQLFREHLL
EQALHCVAQPSPGPGSADGDREFSEALGFLQLLNSCCDTAGAPACSFSIASSTAATAGTDPVAKWWASLTAVVTHWLGRDE
EAAERLYPLVEHLPRALQESERPLPRAALHSFKAARALLGRGKAESGSASLAMCEKASGYLQDSLAATPAGSSIDKAMQLL
LCDLLLVARTSLWQRQKPPPPSQASQGSSSGAQASALELRGFQRDLSGLRRLAQSFRPAMRRVFLHEATARLMAGASPART
HQLLDRSLRRRAGPCGRGGAAAAAAAELEPRPTRREQAEALLLASCYLPPGFLSAPGQRVGMLAEAARTLEKLGDRRLLHD
CQQMLMRLGGGTTVTSS。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 200410017860 CN1279167C (en) | 2004-04-22 | 2004-04-22 | Gene oder of directivity and differentiation factor 1 fat cell of pigs |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 200410017860 CN1279167C (en) | 2004-04-22 | 2004-04-22 | Gene oder of directivity and differentiation factor 1 fat cell of pigs |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1563380A true CN1563380A (en) | 2005-01-12 |
CN1279167C CN1279167C (en) | 2006-10-11 |
Family
ID=34479199
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN 200410017860 Expired - Fee Related CN1279167C (en) | 2004-04-22 | 2004-04-22 | Gene oder of directivity and differentiation factor 1 fat cell of pigs |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN1279167C (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102031304B (en) * | 2010-11-04 | 2012-07-04 | 徐州师范大学 | Single nucleotide polymorphism (SNP) of ADD1 gene of cattle and detection method thereof |
-
2004
- 2004-04-22 CN CN 200410017860 patent/CN1279167C/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
CN1279167C (en) | 2006-10-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1170941C (en) | Human BMP-7 promoter and method for exploring bone-related substance by using the same | |
CN1807623A (en) | Positive regulation gene for cell cycle and its coded protein and uses | |
CN1279167C (en) | Gene oder of directivity and differentiation factor 1 fat cell of pigs | |
CN1206359C (en) | High efficiency retroviral vector which contains genetically engineered cellular non-coding sequence harboring splicing acceptor | |
CN1757732A (en) | CDNA sequence of coding sweet potato phytoene synthetase | |
CN1908171A (en) | Amylose content control gene DU1 of rice endosperm and application thereof | |
CN1524122A (en) | Isocitrate dehydrogenase, gene thereof, and use of the same in the treatment of obesity, hyperlipidemia, and fatty liver in lipid biosynthesis | |
CN100339479C (en) | Gene participating in the synthesis of brassinosteroid | |
CN101062942A (en) | Aspergillus fumigatus original active oxygen lethality related protein and its coding gene | |
CN1295340C (en) | Method for extablishing polymorphism adenoid tumour mouse model | |
CN1301266C (en) | High molecular wheat glutelin subunit, genes encoding same and use thereof | |
CN101054583A (en) | Gene sequence of penaeus monodon cell periodic protein B | |
CN1687414A (en) | Genes in gene gorup of decoupling protein 3 of pig and application thereof | |
CN100344759C (en) | cDNA sequence of dehydrolysis responding transcription factor DREB gene of wheal | |
CN1563379A (en) | Gene order of b subtype of 4 receptor of hydroxytryptarmine in pig | |
CN1563386A (en) | Reducing enzyme protein coded sequence of sulfoxide methionine of cotton | |
CN1280415C (en) | Wheat disease-resistant gene Lr-L1 and its use | |
CN1600861A (en) | Protein coded sequence of binding factor in ethane response element of cotton | |
CN1699567A (en) | Protein encoding sequence of pig nuclear receptor coactivity factor 1 gene | |
CN1225556C (en) | Recombinant nucleic acid of coded human osteogenic protein-1 and its application | |
CN1928084A (en) | Nucleotide and amino acid sequence of gene of penaeus monodon heat shock protein 90 | |
CN1266273C (en) | Promoter for regulating and controlling mouse orphan nucleus recepter and its use | |
CN1740319A (en) | Lefteye flounder MYOSTATIN gene order | |
CN1528889A (en) | Chinese cabbage 9-cis-carotene epoxide dioxygen zymoprotein coding sequence | |
CN1712532A (en) | Pig tyraminase beta protein coding sequence |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C17 | Cessation of patent right | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20061011 |