CN102703444A - Radix polygonati officinalis microsatellite DNA (deoxyribonucleic acid) molecular marker - Google Patents
Radix polygonati officinalis microsatellite DNA (deoxyribonucleic acid) molecular marker Download PDFInfo
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Abstract
The invention discloses a radix polygonati officinalis microsatellite DNA molecular marker, comprising the steps of constructing a radix polygonati officinalis microsatellite (CT) n enrichment library; screening and sequencing microsatellite sequence positive clone; obtaining 55 clones containing the microsatellite repeated sequence; and screening to obtain 9 high polymorphic microsatellite molecular markers Po1, Po2, Po3, Po4, Po5, Po6, Po7, Po8 and Po9. The radix polygonati officinalis microsatellite DNA molecular marker can be used for researching genetic diversity of the radix polygonati officinalis, variation and evolutionary relationship between polygonatum plants and population, has good repeatability and is a reliable and efficient molecular marker.
Description
Technical field
The present invention relates to molecular marking technique, be specifically related to a kind of radix polygonati officinalis micro-satellite molecule genetic marker.
Background technology
Little satellite (Microsatellite) be called again the repetition of short polyphone (Short Tandem Repeats, STRs), simple repeated sequence (Simple Sequence Repeat, SSRs), SSLP (SSLP).Be meant that is dispersed throughout (being called core sequence, being generally 1～6bps) the simple series connection repetition for repeating unit's composition by several nucleotide pairs in the biological gene group.Its multiplicity is an alterable height between the different genotype of same species; But this section multiple two ends are conservative relatively single-copy sequences; Design primer in view of the above, can analyze the variation of core sequence multiplicity, between different genotype, detect polymorphic through round pcr.According to the formation of repeating unit, the microsatellite DNA sequence is divided into 3 types: single type (pure), compound (compound) and discontinuous form (interrupted).For example:
Single type: ATATATATATATATATATATATAT
Compound: ATATATCACACACACACACAC
Discontinuous form: ATATATCA ATATATCA ATATATA
As a kind of molecule marker, microsatellite DNA has following characteristics: be distributed widely in the Eukaryotic genome, in per according to estimates 6 kb length 1 microsatellite locus just arranged; The multiplicity of core sequence is the height variability between individuality, the polymorphism information capacity is high: be codominance, follow the heredity of broad-mouthed receptacle for holding liquid Dare rule; Select neutrality or the like.
Based on above characteristics, microsatellite DNA is widely used in the research of analysis, assistant breeding, structure finger printing, genetic diversity and aspects such as spore and sibship of structure, the assignment of genes gene mapping, cultivar identification and quality saving, the quantitative character gene of plant genetic collection of illustrative plates.
Radix polygonati officinalis has another name called the tail ginseng, beautiful ginseng, and the small bell dish, ground tube and sweet grass are under the jurisdiction of Liliaceae (Liliaceae L.) Polygonatum (Polygonatum Mill.), are per nnial herb, and happiness is cloudy gives birth to, and is distributed widely in area, temperate zone, Eurasia.
The radix polygonati officinalis subterraneous stem can be used as medicine, and its medicinal history has exceeded more than 2,000 year in China.Traditional Chinese medicine thinks that radix polygonati officinalis property is sweet, is slightly cold, attaches to the lung and stomach meridians, can Yin nourishing and lung moistening, reinforcing stomach reg fluid is used for scorching impairment of yin, the disease of thermal burn stomach-Yin.Modern study proof radix polygonati officinalis contains polysaccharide and multiple extract, is delaying senility, and strengthening immunity suppresses tumour, and aspects such as promotion lymphocyte transformation have certain effect.
Since among the people dig excessively serious, in addition the natural crude drugs cycle long, the radix polygonati officinalis wild resource is exhaustion day by day.In fact, radix polygonati officinalis medicine source is comparatively complicated, and little satellite helps to identify the radix polygonati officinalis strain, and distinguishes the pseudo-article of radix polygonati officinalis.Simultaneously, radix polygonati officinalis a series of problems occurred in the cultivation domestication process, and is low like breeding coefficient, even utilize microsatellite marker to the research of radix polygonati officinalis molecular breeding seldom.
Summary of the invention
The technical problem that the present invention solves: the microsatellite DNA molecule marker of separating clone radix polygonati officinalis; Set up radix polygonati officinalis satellite DNA technical system and carry out the radix polygonati officinalis genetic diversity with these molecule markers; The research of Polygonatum platymiscium genetic affinity, and carry out the Polygonatum Idioplasm identification.
The technical scheme of technical solution problem of the present invention: radix polygonati officinalis microsatellite DNA molecule marker, comprise the structure of the little satellite of radix polygonati officinalis (CT) n enriched library, the screening of microsatellite sequence positive colony and order-checking obtain 55 clones that contain little satellite Tumor-necrosis factor glycoproteins.Obtain 9 microsatellite molecular marker Po1, Po2, Po3, Po4, Po5, Po6, Po7, Po8, Po9 that polymorphum is high; Po1 is 360 Nucleotide, and Po2 is 471 Nucleotide, and Po3 is 447 Nucleotide, and Po4 is 408 Nucleotide, and Po5 is 463 Nucleotide, and Po6 is 497 Nucleotide, and Po7 is 424 Nucleotide, and Po8 is 509 Nucleotide, and Po9 is 342 Nucleotide.
Description of drawings
Fig. 1: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po1 site dyes PAGE figure.
Fig. 2: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po2 site dyes PAGE figure.
Fig. 3: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po3 site dyes PAGE figure.
Fig. 4: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po4 site dyes PAGE figure.
Fig. 5: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po5 site dyes PAGE figure.
Fig. 6: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po6 site dyes PAGE figure.
Fig. 7: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po7 site dyes PAGE figure.
Fig. 8: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po8 site dyes PAGE figure.
Fig. 9: the silver of the DNA of 24 radix polygonati officinalis of primer amplified in Po9 site dyes PAGE figure.
Embodiment
Below in conjunction with embodiment the present invention is done detailed explanation.
Embodiment 1:
The structure of radix polygonati officinalis microsatellite DNA enriched library
1) genomic extraction and enzyme are cut:
The CTAB method of introducing with Doyle J (Doyle J. DNA protocols for plants-CTAB total DNA isolation [A]. In Hewitt GM; Johnston A (eds.); Molecular Techniques in Taxonomy [M]. Berlin:Springer; 1991,283-293) extract genome.Cut genome with restriction enzyme Sau 3AI enzyme.Behind 2% the agarose gel electrophoresis, under UV-light, downcut the dna fragmentation of 300-900 bp.Reclaiming test kit with glue reclaims.
2) add joint:
The product that glue is reclaimed behind the purifying adds joint:
oligo?A(5’-GGCCAGAGACCCCAAGCTTCG-3’)
oligo?B?(5’-PO
4-GATCCGAAGCTTGGGGTCTCTGGCC-3’),
Under T4 DNA Ligase effect, spend the night in 16 ℃ of water-baths connections.
3) the PCR detection tabs connects:
With oligo A is primer, is that template is carried out pcr amplification to connect product, (50 μ l reaction systems are following: 25 μ l GoTaq Green Master Mix, and 5 μ l primer oligo A, 5 μ l connect product, add water and mend to 50 μ l.The PCR response procedures: 94 ℃ of preparatory sex change 5 minutes, 94 ℃ of sex change 50 seconds, 56 ℃ of annealing one minute, 72 ℃ were extended 2 minutes, sex change, annealing, three step cycle of extension 25 times, last 72 ℃ were fully extended 10 minutes.After agarose gel electrophoresis 2% detected, the PCR product carried out purifying with purification kit.
4) enrichment with magnetic bead:
Dna fragmentation sex change behind the purifying is formed after the strand and the little satellite probe hybridization that contains vitamin H, form " little satellite probe comprises the genomic dna strand of little satellite " complex body.Hybridization solution is added the magnetic bead of having handled well, and room temperature left standstill 30 minutes.Centrifuge tube is placed on the magnetic force frame, inhales and remove supernatant, use 6 * SSC successively, 2 * SSC, 1 * SSC washes twice under 60 ℃, use at last under 0.1 * SSC room temperature and wash once.Add the TE damping fluid, 95 ℃ of sex change are 15 minutes on the PCR appearance, collect supernatant.Repeat twice.
5) PCR enrichment:
With the enrichment with magnetic bead product is template, and oligo A is that primer carries out pcr amplification, and 25 μ l reaction systems are following, 12.5 μ l GoTaq Green Master Mix, and 1.5 μ l primer oligo A, the DNA after the 2.5 μ l enrichments adds water and mends to 25 μ l.The PCR response procedures is following: 94 ℃ of preparatory sex change 3 minutes, and 94 ℃ of sex change 35 seconds, 60 ℃ of annealing property 35 seconds, 72 ℃ were extended 1 minute, sex change, annealing, three step cycle of extension 25 times, last 72 ℃ were fully extended 12 minutes.The PCR product carries out purifying with purification kit after 2% agarose gel electrophoresis detection.
6) preparation of competent cell:
The single bacterium colony of E.coli DH-5 α is inoculated in the LB substratum that does not contain Amp of 5 ml on the picking flat board, and 37 ℃ of shaking culture are spent the night.Get 1 ml nutrient solution in the LB of 50 ml liquid nutrient medium, 37 ℃ of shaking culture, 2～4 h are to logarithmic phase.Survey OD (0.2～0.6).The nutrient solution branch is filled in the EP pipe (every pipe 1.5 ml bacterium liquid) water-bath 10～15 min.4 ℃ centrifugal, 4000 rpm, 10 min (earlier cooling after centrifugal again) abandon supernatant.Abandon three pipe and pipes behind the supernatant, and ice bath 30 min (first pipe adds 0.1 mol/L, and 1 ml is the CaCl2 of precooling in the ice in advance, mixing, sucking-off to the second pipe is so with three pipes and a pipe).Take out the back gradient centrifugation, 4 ℃, 4000 rpm, 5～8 min.Abandon supernatant.Add 200 μ l precooling glycerine (15%)/CaCl
2(0.1 M) mixing ,-80 ℃ of preservations.
7) connect conversion:
The purified product of getting after the PCR enrichment connects into TA cloning vector (pMD19-T Simple Vector), and 16 ℃ of water-baths connect 2-3 hour.Take out frozen pipe, 10 min on ice recover; The connection product of 10 μ l is added in the 200 μ l competent cells mixing, ice bath 30 min; 42 ℃ of water-bath heat-shocked 90 s; Ice bath 2 min add 600 μ l LB substratum, 37 ℃ of shaking culture 1.5 h; Get 70 μ l nutrient solutions, splash on the LB/Amp/X-gal/IPTG flat board and coating even (X-Gal needs lucifuge); 37 ℃ of forwards of flat board are placed cultivation 0.5h, in 37 ℃ of baking ovens, be inverted overnight cultures then.Through the cultivation of 12 h, treat that the bacterium colony on the flat board is of moderate size, take out flat board and locus coeruleus is fully manifested in 4 ℃ of placements.With white mono-clonal bacterium colony on the toothpick picking flat board of the bacterium of going out, place 96 deep-well plates (deep-well plates has added the LB/Amp substratum that contains the Amp resistance of 400 μ l in advance).After the single bacterium colony picking of white finishes, with non-setting adhesive orifice plate is built the back in 37 ℃, 225rpm shaking culture 4 h.
8) screening of positive colony:
To connect the bacterium liquid that transforms gained is template, utilizes primer oligo A and primer (CT)
12Carry out pcr amplification.15 μ l PCR reaction systems comprise: rTaq enzyme 0.375 U, 10 * Buffer, 1.5 μ l, dNTP 0.2 μ M, MgCl
21.5 μ M, and primer 0.4 μ M (oligo A, (CT)
12), water, template.Response procedures: 94 ℃ of preparatory sex change 3 minutes, 94 ℃ of sex change 50 seconds, 56 ℃ of annealing 35 seconds, 72 ℃ were extended 1 minute, sex change, annealing, three step cycle of extension 30 times.Last 72 ℃ were fully extended 12 minutes.1.5% agarose gel electrophoresis detects.
9) positive colony order-checking and design of primers:
Choose the amplification fragment length and contain the bacterium liquid of positive colony, send the order-checking of the biological (Shanghai) Co., Ltd. of living worker, obtain 51 sequences that contain microsatellite DNA altogether at 300-900 bp.Utilize software Primer Premier 5.0 sequences to design primer according to the microsatellite DNA two ends.
10) screening has the primer of polymorphum:
Extract the genome of radix polygonati officinalis with above-mentioned CTAB method.With primer (table 1) the amplification gene group that designs.Response procedures: 94 ℃ of preparatory sex change 4 minutes, 94 ℃ of sex change 45 seconds, corresponding annealing temperature 30 seconds, 72 ℃ were extended 35 seconds, sex change, annealing, three step cycle of extension 35 times.Last 72 ℃ were fully extended 8 minutes.1.5% agarose gel electrophoresis detects.The PCR product detects with the PAGE electrophoresis.Obtain 9 microsatellite markers that polymorphum is arranged altogether.Po1, Po2, Po3, Po4, Po5, Po6, Po7, Po8, Po9; Po1 is 360 Nucleotide, and Po2 is 471 Nucleotide, and Po3 is 447 Nucleotide, and Po4 is 408 Nucleotide, and Po5 is 463 Nucleotide, and Po6 is 497 Nucleotide, and Po7 is 424 Nucleotide, and Po8 is 509 Nucleotide, and Po9 is 342 Nucleotide.
11) interpretation of result:
Use Cervus3.0 computed in software expectation heterozygosity and observe heterozygosity.Use the value of Genepop3.4 computed in software Hardy-Weinberg and linkage disequilibrium, describe the characteristic of 9 radix polygonati officinalis microsatellite DNA polymorphic sites with this.Can find out by accompanying drawing 1-12; The variety that the length of 9 microsatellite sequences of the present invention all occurs in 21 radix polygonati officinalis that supply examination; This shows that these 9 microsatellite markers of the present invention can be used to study the genetic diversity of radix polygonati officinalis, between the Polygonatum plant species and variation between population and evolutionary relationship; Having very high repeatability, is a kind of reliable and effective molecule marker.
Table 1 primer property list:
F representes upstream primer in the table 1, and R representes. downstream primer.
Claims (2)
1. radix polygonati officinalis microsatellite DNA molecule marker, it is characterized in that: the label of said microsatellite marker is respectively: Po1, Po2, Po3, Po4, Po5, Po6, Po7, Po8, Po9;
Its nucleotide sequence is respectively:
Po1：
gatcacatgc?cgatagttcc?tcggtcacct?agcatattta?gtgtcagtct?tctagctggc 60
gaataaaaac?actaagtcga?aatgaactaa?ggaaagtgct?cttctaactg?tgagagagag 120
agagagagag?agagagagag?agagagagag?agatgttggt?ccatccctca?tggtgtactg 180
tggatttcat?tttgcgctct?ctagctaatt?ggattttagt?cagatttttc?tagttagttt 240
agcaccactc?ataggtagag?ctgtaaaatg?aactgagttt?tggagtgttt?gtgttcgttc 300
aataatttac?acgagtttaa?atttagctga?ttctatatga?acccaaacaa?atactcaatt 360
Po2：
gatcacagat?catgatctgg?tcaaccagaa?acccccgtgt?ctgaccctga?ggatacagat 60
accctctgcc?tggtcacagg?acctgggttg?gttggccaga?cagtgcccag?gtcgaccccg 120
acctattttg?ggctgttatt?ggttttcttt?ggtgttagta?ggttttgttg?gtttcttatt 180
gggtttttga?cttatataac?tataacctac?ttcatctaag?gtttagattt?gttttgtttt 240
attttggatg?gttgagatta?gactttaggt?gggtgtttcc?ctatatatac?catctccaaa 300
ccctagaacc?cattcctcct?cctcctcctc?tctctctctc?tctctctctc?tctctctctt 360
gagcaactct?ctttaagagc?tagggcagag?aatttagggg?aagatttggg?atttttgctg 420
aggttttctt?cgtggatttg?caagcttcgc?atttctaagg?caagtattca?t 471
Po3：
gatcatgcat?atttgtttat?agctagcttc?ccatcaacgt?acgtccgttg?catttgattt 60
gattttcaaa?gtattaggac?ttgaacgtga?agtgagggtg?ggaactggga?actaggactc 120
ggatgagaga?gagagagaga?gagagagaga?gagagagaga?gagagagaga?tatgctgcag 180
gaacttgtgg?ataaatagag?atatgtgtgg?cactctctca?ttgattatac?ttagttcact 240
ctctcgaaac?gccacagaga?tagacaaagg?cttgattgtt?agatcagcat?acagatggca 300
agaaatggca?gcggttatgc?ttgctagttt?ggtgagtggt?gagacatttt?gtcctctctg 360
gccgtcgtac?ttgctataaa?gacgacgtgg?aaaggcccga?ccacgagaat?gccgaccgca 420
tcgtttttcg?acgataatgt?tgttttt 447
Po4：
gatcacgaac?ttggaagctt?cgactttggt?gattgttgga?gtgagctaca?aattgaagag 60
ggtgggtttg?cttgagctca?tgaataagag?gaagggagga?gcttggcttt?ctcttgaaaa 120
aggaatgaga?atgaaaagag?agagaggggg?aggaattgtt?tttatcaagg?gggcatggtg 180
cattaaatgc?ttggaatgat?ggagtgctgg?gtattaaatg?gaggagggag?agagagagag 240
agagagagag?agagagagag?agagagagag?agagagagag?agagagagtg?atgggacttt 300
tctcatccat?gaatgagaca?aagttaacaa?gagggatgtt?aacaattcaa?actggtaaaa 360
cacatgagtg?aaccaaacca?gtccacgaat?cgcagtaatt?cgactgtg 408
Po5：
gatcttgtgc?caacctcgct?attaaactcc?ttctttagct?cgtaacaaca?ccattgctct 60
tccactactc?aatctacgcc?ttcactgtgt?ccctactttc?tagcctcacc?ttctctctct 120
ctctctctcc?ctctctctct?ctctctctct?ctctctctct?ctctctcttc?cgttcgcctc 180
cgggcatttc?tttagccacc?cggcctccct?ttccgccttc?cgggctactc?tttacctccg 240
agctatggtt?acttcttaca?agctggcttt?cgggccctcg?tttagcactt?ttagtgcttt 300
tagccttcag?gcttagcttc?ggttactttc?acgagttgac?tctcgggtcc?tcgtttagca 360
cttttagtgc?ttttagcatt?cgtacttgct?ttccagcctc?gcaagagcaa?aggaggtgct 420
gccaaatttt?ctgctgatac?tagactgcag?cctaactaca?atg 463
Po6：
gatcaacatc?tccctcacct?ttttgcagtg?gtttccaagg?gcctaaggcc?caatctttcg 60
ctggctcagt?gaagttaagc?ccaagcttgt?ctttttgcag?tggtttccat?atgtgaagtt 120
aagcccaagc?ttgtcaacat?ctccctaata?tatatagccc?aatctatgta?ggcctgccta 180
gtgggctgaa?ttatcctacc?acattactcg?ctcatattcc?accaattccg?tatgtggact 240
gattcagcac?cacccaaaac?ccgacacgcc?tccttcgctt?ctccaccaca?tccacctcca 300
cgtgtcggtt?tcccccctcc?cttctctgtt?tcaatattaa?ctcctttttc?tgtccgaaac 360
aaaacccatc?tcttttctct?ctctctctct?ctctgtgtgt?gttcagtgtt?caaggtctca 420
acaaagagag?tagcaatggc?gacttcttct?tcttcttctt?cgactagtcc?gctgctgaag 480
tacgagctgg?atatcgt 497
Po7：
gatcatcggc?atcgtcgtca?agagggatgg?agagcgtctc?tctcgccttc?ggacgctgat 60
agcagagaga?cccatcgatg?gagcaacaaa?ggaaaacaac?caacaaaatc?acaacgaaac 120
gaagattact?tgcttcttcc?atcatgaatg?agtatgaagt?ttaatgtgga?atactgtgag 180
agagagagag?agagagagag?agagagagag?agagagagag?agagagagag?agagaggaga 240
gataatggag?gctgcaggca?caatttataa?tgtggtattt?acaaagaaag?aaagaaatta 300
gattttttga?gcgtttgatt?tgaaaaaaat?tgcaacactc?tctccttcta?tctcttccta 360
catttgcatg?cccatttagt?tttcggtttg?ggacacgagg?gggccttcat?aagttgactc 420
gtgt 424
Po8：
gatcatccgg?aatcctgcta?gttggccttg?gattggaccg?gtgtccgtta?ccggatccaa 60
tattgatatc?tgtacccgtt?tctgtgttta?atatagtgga?ttgttaattt?gaactgtgtt 120
aatacagttt?cctcttgtct?ttgatggcat?ggcaagagcc?ccaccttcct?ttgtctaatc 180
aaagccccat?ccccttgcat?ttaatacttg?ttattcactc?tctctctctc?tctctctctc 240
tctctctctc?ccatttttta?cctcctccct?tcccttctct?ctttctcttc?tcatcttcaa 300
agagcaacct?aagggaagca?agagaacacc?accacaccaa?gagctcttcc?aagcctcccc 360
ttgctccggt?gaacactccg?gtaagctccg?gccgagctcc?gacgaactta?ggcgagtgcc 420
atgctttcct?tagcctacgg?actgtaaatt?gtgagctcac?gagttcaaga?aatctctctt 480
taaaagagaa?gccttaaggg?ctattaatt 509
Po9：
cccttaagcc?ctcattgctg?cccctgacca?agccctcgtc?tcgccggccg?accggtcagc 60
tgagtaatgg?ccaacctggt?tagtcgaccg?acctcctgct?cccaatagag?cgccgcctct 120
ctctctctct?ctctctctct?ctctctctct?ctcttttcaa?atttccccac?actccgctta 180
attcttctcc?taactccctg?tcactcccta?actctcccta?attcccaccc?taactccctc 240
tatctctctc?actttctctc?ctaactctct?cgatctcgcc?tctaacctat?ctgagctccg 300
gacaactggc?atctccacca?tattctcttt?cactcctcca?tt 342。
2. a kind of radix polygonati officinalis microsatellite DNA molecule marker according to claim 1 is characterized in that:
The primer sequence in described Po1, Po2, Po3, Po4, Po5, Po6, Po7, Po8, nine sites of Po9 is:
Po1-F GAA?CTA?AGG?AAA?GTG?CTC?TTC?TAA?C
Po1-R CTC?TAC?CTA?TGA?GTG?GTG?CTA?AAC
Po2-F TTG?AGA?TTA?GAC?TTT?AGG?TGG?GTG
Po2-R TGA?ATA?CTT?GCC?TTA?GAA?ATG?CG
Po3-F CTT?GAA?CGT?GAA?GTG?AGG?GTG
Po3-R CTG?CCA?TTT?CTT?GCC?ATC?TGT?AT
Po4-F ATG?CTT?GGA?ATG?ATG?GAG?TGC?T
Po4-R GAT?TCG?TGG?ACT?GGT?TTG?GTT?C
Po5-F CTA?CGC?CTT?CAC?TGT?GTC?CCT
Po5-R CTC?GGA?GGT?AAA?GAG?TAG?CC
Po6-FGCT?TCT?CCA?CCA?CAT?CCA?CCT
Po6-R GTC?GCC?ATT?GCT?ACT?CTC?TTT?GT
Po7-FGAG?CAA?CAA?AGG?AAA?ACA?ACC?AAC
Po7-R?GTG?TCC?CAA?ACC?GAA?AAC?TAA?ATG
Po8-F AGT?TTC?CTC?TTG?TCT?TTG?ATG?G
Po8-R GCA?CTC?GCC?TAA?GTT?CGT?C
Po9-F GCA?CTC?GCC?TAA?GTT?CGT?C
Po9-R CAG?ATA?GGT?TAG?AGG?CGA?GAT。
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020033336A (en) * | 2000-10-30 | 2002-05-06 | 박선민 | Food composition comprising extract from Polygonatum odoratum (Mill.) Druce |
| CN1363371A (en) * | 2002-01-11 | 2002-08-14 | 潘文光 | Multifunctional bio-active medicine for preventing and curing AIDS |
| KR20040024008A (en) * | 2002-09-12 | 2004-03-20 | 선만규 | Health food composition for improving constipation |
| KR20040058668A (en) * | 2002-12-27 | 2004-07-05 | 한국화장품주식회사 | Polygonati Rhizoma extract having skin whitening effect and skin whitening cosmetic composition containing the same |
| CN1513968A (en) * | 2002-12-31 | 2004-07-21 | 李玉文 | Special function wine using nucleotide Chinese herbal medicine as main raw material |
| CN1557842A (en) * | 2004-02-02 | 2004-12-29 | 中南大学湘雅二医院 | Process for extracting polygonatum polysaccharides, preparing process for medical preparation and use thereof |
| KR100702901B1 (en) * | 2006-12-24 | 2007-04-09 | 서희동 | A manufacturing method of makgeolli containing medicinal properties which used deep sea water |
| CN1974598A (en) * | 2005-07-26 | 2007-06-06 | 香港中文大学 | Isolated proteins from a traditional Chinese medicine yuzhu and use thereof |
-
2012
- 2012-05-25 CN CN 201210165286 patent/CN102703444B/en not_active Expired - Fee Related
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020033336A (en) * | 2000-10-30 | 2002-05-06 | 박선민 | Food composition comprising extract from Polygonatum odoratum (Mill.) Druce |
| CN1363371A (en) * | 2002-01-11 | 2002-08-14 | 潘文光 | Multifunctional bio-active medicine for preventing and curing AIDS |
| KR20040024008A (en) * | 2002-09-12 | 2004-03-20 | 선만규 | Health food composition for improving constipation |
| KR20040058668A (en) * | 2002-12-27 | 2004-07-05 | 한국화장품주식회사 | Polygonati Rhizoma extract having skin whitening effect and skin whitening cosmetic composition containing the same |
| CN1513968A (en) * | 2002-12-31 | 2004-07-21 | 李玉文 | Special function wine using nucleotide Chinese herbal medicine as main raw material |
| CN1557842A (en) * | 2004-02-02 | 2004-12-29 | 中南大学湘雅二医院 | Process for extracting polygonatum polysaccharides, preparing process for medical preparation and use thereof |
| CN1974598A (en) * | 2005-07-26 | 2007-06-06 | 香港中文大学 | Isolated proteins from a traditional Chinese medicine yuzhu and use thereof |
| KR100702901B1 (en) * | 2006-12-24 | 2007-04-09 | 서희동 | A manufacturing method of makgeolli containing medicinal properties which used deep sea water |
Non-Patent Citations (5)
| Title |
|---|
| 刘婷婷: "长梗黄精11个微卫星位点的筛选及跨种扩增", 《全国优秀硕士论文全文数据库 农业科技辑》, 15 February 2012 (2012-02-15), pages 047 - 186 * |
| 周晔等: "ISSR法鉴定中药玉竹与小玉竹", 《中医药学报》, vol. 34, no. 5, 31 October 2006 (2006-10-31), pages 7 - 11 * |
| 李钟: "用RAPD技术鉴别湖南道地药材玉竹", 《湖南中医药大学2003年度学位论文集》, 31 December 2003 (2003-12-31) * |
| 林丽美: "RAPD技术在商品药材玉竹鉴定中的应用", 《中国优秀硕士论文全文数据库 医药卫生科技辑》, 15 May 2010 (2010-05-15), pages 057 - 1 * |
| 潘清平等: "玉竹不同品种的ISSR分子鉴定", 《中国现代中药》, vol. 10, no. 10, 31 October 2008 (2008-10-31), pages 28 - 30 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498482A (en) * | 2014-11-28 | 2015-04-08 | 安徽师范大学 | Pteroceltis tatarinowii microsatellite DNA molecular marker and isolation and screening method thereof |
| CN104498482B (en) * | 2014-11-28 | 2018-04-13 | 安徽师范大学 | Wingceltis Microsatellite DNA molecular marker and separating screening method |
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