CN1502231A - Method for obtaining inbred line small size pig - Google Patents
Method for obtaining inbred line small size pig Download PDFInfo
- Publication number
- CN1502231A CN1502231A CNA021490309A CN02149030A CN1502231A CN 1502231 A CN1502231 A CN 1502231A CN A021490309 A CNA021490309 A CN A021490309A CN 02149030 A CN02149030 A CN 02149030A CN 1502231 A CN1502231 A CN 1502231A
- Authority
- CN
- China
- Prior art keywords
- inbred line
- generation
- inbreeding
- miniature pig
- pig
- 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
- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000009399 inbreeding Methods 0.000 claims abstract description 24
- 244000309715 mini pig Species 0.000 claims abstract description 19
- 108090000623 proteins and genes Proteins 0.000 abstract description 11
- 241001465754 Metazoa Species 0.000 abstract description 8
- 230000000052 comparative effect Effects 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 5
- 239000003814 drug Substances 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 238000009395 breeding Methods 0.000 abstract 2
- 230000001488 breeding effect Effects 0.000 abstract 2
- 238000011160 research Methods 0.000 description 15
- 241000282898 Sus scrofa Species 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- 108020004414 DNA Proteins 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 8
- 238000003752 polymerase chain reaction Methods 0.000 description 7
- 230000002068 genetic effect Effects 0.000 description 6
- 108091092878 Microsatellite Proteins 0.000 description 5
- 238000010171 animal model Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 108700028369 Alleles Proteins 0.000 description 3
- 108020004705 Codon Proteins 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000037308 hair color Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 230000035479 physiological effects, processes and functions Effects 0.000 description 3
- 241000287828 Gallus gallus Species 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 235000013330 chicken meat Nutrition 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 230000001568 sexual effect Effects 0.000 description 2
- 241000208140 Acer Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 229920005601 base polymer Polymers 0.000 description 1
- 230000037148 blood physiology Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 244000144987 brood Species 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000011173 large scale experimental method Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- GJAWHXHKYYXBSV-UHFFFAOYSA-N quinolinic acid Chemical compound OC(=O)C1=CC=CN=C1C(O)=O GJAWHXHKYYXBSV-UHFFFAOYSA-N 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to a method for obtain inbred line miniature pig. The color gene of the miniature pig produced by said method does not be separated, said miniature pig can be used as qualified experimentol animal for comparative medicine. Said invention technical scheme is as follows: using wu zhishann miniature pig to make of full-sib, offspring-parent or grandparent and grandchild three generations inbreeding breeding for above 8 generations, in order to obtain better effect, perfectly, making inbreeding breeding for above 10 generations. It can meet the requirements for life science development.
Description
Technical field
The present invention relates to a kind of method that obtains the inbred line animal, particularly relate to a kind of method that obtains the inbred line miniature pig.
Background technology
Inbred line laboratory animal gene high homogenous, precision instrument that can be in physics research and the analytical reagent in the chemical reaction are for biomedical and whole life science provide experimental result sensitive, that deviation is minimum.Along with the research and development of inbred mouse, rabbit and inbred line chicken are used, the developing and solved many-sided many new difficult problems such as mammal heredity, growth, physiology, immeasurable effect has been played in the development of biology, life science especially physianthropy.Since C.C.Little (1909) cultivates inbred line DBA mouse, 388 of inbred mouses, 34 of inbred line rabbit, 40 of inbred line chickens have been cultivated at present in the world, to satisfy the demand of scientific research.But because the large-scale experiment animal breeds all many-sides such as characteristic, genetic development at physiology different with the small-scale test animal, very difficulty is bred in inbreeding, though experienced long research course, obtained some progress (Wessea, 1950, J.F.Lasley, 1978, Li Dengyuan etc., 1982), but do not see successfully report so far.
Great deal of research results shows both at home and abroad, pig and people's dissect physiology, vascular system, digestive system, skeleton development, Nutrition and Metabolism etc. have bigger similitude, very approaching such as people and pig blood pH, erythrocyte number, red blood cell diameter, be respectively 0.737-0.745,0.736-0.749; 4.0-5.5 * 10
12/ L, 5.0-7.0 * 10
12/ L; 7.1 micron, 6.1 microns.Thereby when having guaranteed in the pig blood input human body, can pass through the capillary of partes corporis humani position smoothly.Thus, the expert reaches common understanding in the following aspects both at home and abroad: pig is people's ideal animal model, xenotransplant donor.The eighties, pig is heightened the existing trend that replaces dog both at home and abroad as the laboratory animal consumption.At present, miniature pig has been widely used in fields such as pharmacy, comparative medicine in the world, and just the cultivation direction towards SPFization develops.Countries in the world have been cultivated 20 kinds of experiments and have been used miniature pig for satisfying various objectives, but all non-inbred variety.
WZSP (Wu Zhishan miniature pig) is had inconvenient traffic in the middle part of originating in Hainan Province, natural enclosing reaches the mountainous regions of tropics of not raising the boar custom.Under this specific ecological conditions, by successive dynasties " match ", " son is joined mother " equal altitudes close relative's procreation mode and natural selection for many years, formed a kind of small, very slow mini-breed of weightening finish, there is the title of " 3 years less than 50 jin " locality, is the excellent material that obtains the inbred line miniature pig.
Summary of the invention
The purpose of this invention is to provide a kind of method that obtains the inbred line miniature pig, the miniature pig hair color gene that utilizes this method to produce does not separate, can be used as the qualified laboratory animal of comparative medicine, satisfies the needs of life science development.
For achieving the above object, the present invention by the following technical solutions: a kind of method that obtains the inbred line miniature pig is WZSP to be carried out full sibs, close son or grandparent and grandchild's three from generation to generation interior inbreeding breed more than 8 generations.
In order to obtain better effect, inbreeding is bred and is preferably in more than 10 generations.
The inbred line miniature pig that utilizes said method production to obtain is named as WZSP, and its colony's inbreeding coefficient is one of strain that the inbreeding coefficient of report is the highest in the world at present up to more than 0.965.The inbred line miniature pig that utilizes method of the present invention to obtain is small, body weight light, very slowly (6 monthly age 13.43+3.27kg grow; 24 monthly age 36.65+9.17kg), early sexual maturity, inheritance stability and energy inbreeding; The multiple blood physiology biochemical values of this inbred line miniature pig is approximate human simultaneously, and the heart coronary artery descending anterior branch distributes closely similar with Chinese's second type (accounting for 90%).WZSP leukocyte classification medium size lymphocyte content (75%) is significantly higher than common pig (52%) and human (25-35%).By HLA serology (SLA) research, find that W2 antigen appears in its C site, W6 antigen appears in the B site, this composition was not all found in other product boar.The inventor is by influencing meat to kind more than 50 and the local flavor specificity substance is discovered, the quinolinic acid of WZSP and malonic acid ethyl pentyl group fat content are higher than 8 times of common porks and 20 times respectively, and microsatellite DNA mark is discovered: SW2,103bp; W70,264bp; SW936,85bp; SW874,189bp; SW61, the 210bp site is the distinctive genetic marker of WZSP, for it has smallly, early sexual maturity, meat wait the germplasm specificity that molecular genetics theoretical foundation is provided well.
By discovering the WZSP molecular genetics is specific: WZSP dna fingerprint figure has extremely strange consistent specificity; Growth hormone gene sequence and common pig (Vize.1990) have very big-difference, that is: there are the disappearance of continuous 14 bases, that is: TGGCAGAGGGCGGG in intron III district; Four include the subarea dislocation of 6 bases are arranged, and they are: G, A, G, A, C, G; The exon district has 3 bases to change, and has caused 3 amino acid residues to change.They are that the 373rd bit base G in the 2nd exon becomes A, and corresponding codon has CGG to become CAG, and amino acid has positively charged polar amino acid Arg to become uncharged polar amino acid Gln; The 1149th bit base A in the 4th exon becomes G, and corresponding codon becomes CGG by CAG, and amino acid becomes charged polar amino acid Arg (Q121R) by uncharged polar amino acid Gln; The 1574th bit base A in the 5th exon becomes G, and corresponding codon becomes GGC by GAC, and amino acid becomes uncharged polar amino acid Gly (D168G) by electronegative polar amino acid Asp.MHSn (stress reaction) gene frequency is 0 (0/44); Separate, clone's 221bp fragment (Landrace, pig does not have in the maple river rising in Ningxia and flowing into central Shaanxi) be its peculiar fragment, it is its specificity genetic marker, this 221bp fragments sequence is measured (can not disclose), and its base sequence is as follows: CATAGGAAGG TTAAT AATTT AGTAT GGTTT TTAAA ATTCC CCTTC CATTT TTATCTACCC ATTTG AAAAT TATAT ATACC TATAC CTTCA CACAG TCCTC CACTGACACA GTTCA TACAT TAATA GCTCT AGTAT ATCTA GAGAG TAGTT TACTGCTGTC ATGTT TATAT ATTCT CTAAT TCTTG GCCAA AAAAA AAAAA AATTACCAAC AGATC AGTAC C.RALP studies confirm that the WZSP inbreeding breeds reliably, heredity height homogeneity.
The inbred line miniature pig that utilizes method of the present invention to obtain is the desirable material of animal and veterinary, Nutrition and Metabolism, embryo engineering, biotechnology, human Comparative Medicine Research, in human Comparative Medicine Research, human xenotransplant basic research, will give play to effect distinctive, difficult to the appraisal.Develop at present, obtained certain progress at aspects such as skin, oral cavity, cardiovascular, liver, kidneys.
Embodiment
Embodiment 1, full-sib mating are produced the inbred line miniature pig
Utilize brood 1 public 1 mother (because of other 1 sow 6 monthly age death), carry out the generation inbreeding, carried out 15 altogether from generation to generation, 0-5 inbreeding from generation to generation nest litter size is the 6-8 head;
6 generation nest litter sizes are 5.5 ± 0.50 (n=3);
7 generation nest litter sizes are 5 ± 2.0 (n=1);
8 generation nest litter sizes are 5 (n=1);
9 from generation to generation the nest litter sizes be 4.5 ± 2.0 (n=4), and stillborn foetus appears, mummy accounts for 33.33% (8/16);
10 generation nest litter sizes are 4.5 ± 0.83 (n=6), stillborn foetus, mummy 7.41% (2/27), and piglet survival ratio is 22.22% (6/27);
11 generation nest litter sizes are 5.09 ± 0.86 (n=11), stillborn foetus, mummy 1.79% (1/56), and piglet survival ratio is 48.21% (27/56);
12 generation nest litter sizes are 5.63 ± 2.50 (n=8), and piglet survival ratio is 46.07% (21/45);
13 generation nest litter sizes are 5.63 ± 0.875 (n=8), and hypoplasia accounts for 4.44% (2/45), and piglet survival ratio is 53.33% (24/45);
14 generation nest litter sizes are 5.75 ± 1.00 (n=6), and piglet survival ratio is 66.67% (22/33);
15 generation nest litter sizes are 4.88 ± 1.86 (n=8), and hypoplasia accounts for 7.69% (3/39), and piglet survival ratio is 66.67% (26/39).
From experimental result as can be seen: 1, inbreeding is bigger to the influence of WZSP fertility; 2, the 9th generation piglet survival ratio lower, along with the increase of inbreeding algebraically, piglet survival ratio improves gradually, stillborn foetus, mummy ratio reduce gradually, its nest litter size also has the trend of increase; 3, assorted hair phenomenon does not appear in 300 remainder WZSP inbreeding offsprings, and its hair color feature shows, has reached the degree of high homogenous through its hair color gene after the inbreeding of 8 generations.
In the above-described embodiments, utilize father and daughter, mothers and sons or grandparent and grandchild's three from generation to generation interior inbreeding to breed, the result is identical.
Embodiment 2, WZSP dna fingerprinting research from generation to generation
The generation dna fingerprinting research that utilizes 33.6,33.15 probes to carry out.Its basic process is at first to randomly draw 8 pigs from WZSP colony, carries out the dna fingerprint analysis with probe 33.6 and 33.15.Each sample all enzyme is cut 8-10 μ g DNA, and electrophoretic separation, alkaline process are transferred on the nylon membrane then.Probe carries out mark by random primer hair α-32pdCTP; Hybridization solution is 0.5mol/LPO4 (PH7.2), 7%SDS, 1mmol/L EDTA (PH8.0); Film is pressed behind the probe hybridization 15hr that 1-2 * 106cpm/ml adds sex change in hybridization solution behind the prehybridization 1-2hr about.Hybridization temperature and to wash the film condition different because of probe.
The result shows that dna fingerprint figure F value between inbreeding the 8th generation individuality is that numerical value between 0.678, the 8 generation and the 9th generation is that numerical value between 0.785, the 9 generation and the 10th generation is that numerical value between 0.855, the 10 generation individuality is 0.906.Show that its inbreeding algebraically conforms to the variation of dna fingerprinting F value; WZSP is along with the increase of inbreeding algebraically, and its dna fingerprinting F value constantly raises, and has proved reliability and accuracy that inbreeding is bred.
Embodiment 3, the research of WZSP inbreeding family gene pure degree
1, STR research
Utilize 18 little satellites that the WZSP inbred line is carried out genetic pure and mild degree and homogeneity is studied.At first carry out the extraction of pig blood sample genome DNA, the PCR selection of primers, determining of PCR reaction condition detects the PCR product again, carries out the polymorphism analysis of microsatellite DNA at last.The characteristic of 18 pig microsatellite DNAs is as follows:
| ???MS | ????Chr | ???????????????????Primer?Sequence |
| ??S0036 | ????2 | ?F：AGTGACGTGAGGGGTCTGCTCCTC??R：ATGGACGGTGGATTCACAGCC |
| ??S0070 | ????10 | ?F：GGCGAGCATTTTCATTACAG?????R：GAGCAAACAGCATCGTGAGC |
| ??S0301 | ????4 | ?F：AACCAAGCCACTTTCCCAC??????R：GCTGAAATGCCCATCTGG |
| ??Sw1119 | ????15 | ?F：CAACCTCAAAAATGGAGAAAGG???R：GTTCTTGCGGTGTTTGGC |
| ??Sw1377 | ????11 | ?F：TTCAAGGTTGGAAAGACAGTCC???R：ATGAGGAGTTTGAACTATTGGG |
| ??Sw1695 | ????2 | ?F：ATAAGGGAAATCAGGGCTGAGC???R：TCCCAGGAGCTACCATATGC |
| ??Sw2 | ????5 | ?F：TGCCAATGGTGTGGCTTATAA????R：CCCTGAAGGCTCAGATGGT |
| ??Sw205 | ????8 | ?F：CACAGGTCCATCACCTCATG?????R：GGGTATCTAATGTACATCACGG |
| ??Sw225 | ????13 | ?F：AGGACCCACCAAGAGTTACC?????R：TGCTGGTAATGGGTGATTGG |
| ??Sw2409 | ????4 | ?F：ACTGGTGTCCTTACAAGAAGGG???R：CTTTGCCATAATCTTCACATGG |
| ??Sw2612 | ????14 | ?F：AGCATTTTGGGGGGATTAG??????R：GGCCTCTCTTCCTGTCCTG |
| ??Sw445 | ????4 | ?F：GCTCCCTGGCACTCATTG???????R：CACACACACAAGCAGGTGC |
| ??Sw510 | ????14 | ?F：GTTCCCATAAGCTCATCACTCA???R：ATATTCTGCACTTGCAGCCC |
| ??Sw61 | ????8 | ?F：GAGAGGGATGAGGACTCTGG?????R：AGAGCATTCCAGGCTCTCA |
| ??Sw71 | ????6 | ?F：GATCACCCTTATCCCCATTC?????R：TAGAAACACCATCATCCATTCA |
| ??Sw874 | ????12 | ?F：AAAAGAACCCAACTACAGCAGC???R：TTTATGAGGGTATCTGACACC |
| ??Sw902 | ????3 | ?F：ATCAGTTGGAAATGATGGGC?????R：CTTGCCTCAAAGAGTTGTAAGG |
| ??Sw936 | ????15 | ?F：TCTGGAGCTAGCATAAGTGCC????R：GTGCAAGTACACATGCAGGGGGG |
The result shows WZSP inbreeding family gene high homogenous, family F14 generation, the average effective allelomorph number of 18 microsatellite locus is 1.8967 (the average effective allelomorph number of common each microsatellite locus of boar is more than 6), the gene pure rate is 0.6888, and heterozygosity is 0.3112 (common boar is 0.53-0.67); And have 2 sites (SW2409, SW445) gene frequency has reached 100% isozygoty, other have 3 sites (SW2, S0070, SW2612, SW71) gene frequency has reached 91.7% isozygoty; SW2 103bp, S0070 264bp, SW93685bp, SW874 189bp and SW61 210bp site are the distinctive genetic marker of WZSP.
2, RAPD research
The RAPD technology is based upon polymerase chain reaction (PCR) (Polymerase Chain Reaction, PCR) on the amplification basis, the oligonucleotides strand (being generally 10 base polymers) that it utilizes a series of different randomized arrangement base books to be permitted is a primer, and research genome is increased.Amplified production separates by agarose gel electrophoresis, thereby detects the polymorphism of amplified production fragment.This research utilizes 36 probes that the WZSP inbred line is carried out genetic pure and mild degree and homogeneity is studied.The sequence of probe is:
| Numbering | Sequence | Numbering | Sequence | Numbering | Sequence |
| ?S1 | ?GTTTCGCTCC | ?S2 | ?TGATCCCTGG | ?S3 | ?CATCCCCCTG |
| ?S4 | ?GGACTGGAGT | ?S5 | ?TGCGCCCTTC | ?S6 | ?TGCTCTGCCC |
| ?S7 | ?GGTGACGCAG | ?S8 | ?GTCCACACGG | ?S9 | ?TGGGGGACTC |
| ?S10 | ?CTGCTGGGAC | ?S11 | ?GTAGACCCGT | ?S12 | ?CCTTGACGCA |
| ?S13 | ?TTCCCCCGCT | ?S14 | ?TCCGCTCTGG | ?S15 | ?GGAGGGTGTT |
| ?S16 | ?TTTGCCCGGA | ?S17 | ?AGGGAACGAG | ?S18 | ?CCACAGCAGT |
| ?S19 | ?ACCCCCGAAG | ?S20 | ?GGACCCTTAC | ?S21 | ?CAGGCCCTTC |
| ?S22 | ?TGCCGAGCTG | ?S23 | ?AGTCAGCCAC | ?S24 | ?AATCGGGCTG |
| ?S25 | ?AGGGGTCTTG | ?S26 | ?GGTCCCTGAC | ?S27 | ?GAAACGGGTG |
| ?S28 | ?GTGACGTAGG | ?S29 | ?GGGTAACGCC | ?S30 | ?GTGATCGCAG |
| ?S31 | ?CAATCGCCGT | ?S32 | ?TCGGCGATAG | ?S33 | ?CAGCACCCAC |
| ?S34 | ?TCTGTGCTGG | ?S35 | ?TTCCGAACCC | ?S36 | ?AGCCAGCGAA |
The result is that each probe obtains the 1-6 band.Confirm that WZSP inbred line allelomorph has reached the degree of high homogenous.
Claims (2)
1, a kind of method that obtains the inbred line miniature pig is WZSP to be carried out full sibs, close son or grandparent and grandchild's three from generation to generation interior inbreeding breed more than 8 generations.
2, method according to claim 1 is characterized in that: described inbreeding was bred more than 10 generations.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021490309A CN1254172C (en) | 2002-11-21 | 2002-11-21 | Method for obtaining inbred line small size pig |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021490309A CN1254172C (en) | 2002-11-21 | 2002-11-21 | Method for obtaining inbred line small size pig |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1502231A true CN1502231A (en) | 2004-06-09 |
| CN1254172C CN1254172C (en) | 2006-05-03 |
Family
ID=34233457
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021490309A Expired - Lifetime CN1254172C (en) | 2002-11-21 | 2002-11-21 | Method for obtaining inbred line small size pig |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1254172C (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100400674C (en) * | 2005-02-28 | 2008-07-09 | 中国农业科学院畜牧研究所 | Molecular marker of Wuzhishan pig and its application |
| CN1871903B (en) * | 2006-06-28 | 2011-09-07 | 吉林省农业科学院 | Method for breed porker by using Changbai mountain wild boar as male parent and Songliao black pig as female parent |
| CN103272274A (en) * | 2013-05-28 | 2013-09-04 | 北京博辉瑞进生物科技有限公司 | Biological repair tablet for herniae and preparation method thereof |
| CN103272275A (en) * | 2013-05-28 | 2013-09-04 | 北京博辉瑞进生物科技有限公司 | Biological repair tablet for endocranium and preparation method thereof |
| CN103923994A (en) * | 2014-04-16 | 2014-07-16 | 中国农业科学院北京畜牧兽医研究所 | Method for identifying Wuzhishan miniature pig inbred line by applying SNP (single nucleotide polymorphism) of MLANA gene |
| CN106086166A (en) * | 2016-06-07 | 2016-11-09 | 苏州贝斯派生物科技有限公司 | Reagent of detection paraffin section DNA mass and preparation method and application |
| CN110122415A (en) * | 2019-05-28 | 2019-08-16 | 上海市农业科学院 | The mating system of height inbreeding Experimental Miniature Pig closed colony |
| CN111926081A (en) * | 2020-07-10 | 2020-11-13 | 中国农业科学院兰州兽医研究所 | Single hair color inbred line cooperative small pig breeding method and genetic auxiliary identification method |
-
2002
- 2002-11-21 CN CNB021490309A patent/CN1254172C/en not_active Expired - Lifetime
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100400674C (en) * | 2005-02-28 | 2008-07-09 | 中国农业科学院畜牧研究所 | Molecular marker of Wuzhishan pig and its application |
| CN1871903B (en) * | 2006-06-28 | 2011-09-07 | 吉林省农业科学院 | Method for breed porker by using Changbai mountain wild boar as male parent and Songliao black pig as female parent |
| CN103272274A (en) * | 2013-05-28 | 2013-09-04 | 北京博辉瑞进生物科技有限公司 | Biological repair tablet for herniae and preparation method thereof |
| CN103272275A (en) * | 2013-05-28 | 2013-09-04 | 北京博辉瑞进生物科技有限公司 | Biological repair tablet for endocranium and preparation method thereof |
| CN103923994A (en) * | 2014-04-16 | 2014-07-16 | 中国农业科学院北京畜牧兽医研究所 | Method for identifying Wuzhishan miniature pig inbred line by applying SNP (single nucleotide polymorphism) of MLANA gene |
| CN106086166A (en) * | 2016-06-07 | 2016-11-09 | 苏州贝斯派生物科技有限公司 | Reagent of detection paraffin section DNA mass and preparation method and application |
| CN110122415A (en) * | 2019-05-28 | 2019-08-16 | 上海市农业科学院 | The mating system of height inbreeding Experimental Miniature Pig closed colony |
| CN110122415B (en) * | 2019-05-28 | 2021-06-08 | 上海市农业科学院 | Breeding method of small pig closed group for high inbred experiment |
| CN111926081A (en) * | 2020-07-10 | 2020-11-13 | 中国农业科学院兰州兽医研究所 | Single hair color inbred line cooperative small pig breeding method and genetic auxiliary identification method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1254172C (en) | 2006-05-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106399523B (en) | A kind of molecular labeling SV193 and its application technology distinguishing Chinese native pig breed and Large White | |
| CN108660161A (en) | Method of the preparation based on CRISPR/Cas9 technologies without mosaic gene knock-out animal | |
| CN103534352A (en) | Method for selecting plasma cells and plasmablasts, method for producing target antigen-specific antibody, and new monoclonal antibody | |
| CN107043782A (en) | A kind of gene knockout method and its sgRNA fragments and application | |
| CN1254172C (en) | Method for obtaining inbred line small size pig | |
| Zhang et al. | De novo transcriptome analysis reveals insights into different mechanisms of growth and immunity in a Chinese soft-shelled turtle hybrid and the parental varieties | |
| CN105063021B (en) | The SNP marker related to label of pig fat deposition description and its application | |
| CN108103108A (en) | The preparation and its application of Cebpa gene delection zebra fish mutant | |
| Liu | Development of genomic resources in support of sequencing, assembly, and annotation of the catfish genome | |
| CN108753837A (en) | The construction method and sgRNA of a kind of hyperlipidemia or rabbit model | |
| CN113913425B (en) | sgRNA combination of targeted AHRR gene and application thereof | |
| CN104351096A (en) | Paramisgurnus dabryanus selective breeding method | |
| CN113005142B (en) | Preparation of mutant zebra fish with doc2 alpha and doc2 beta gene deletion and construction method of zebra fish autism model | |
| CN104928386B (en) | Molecular labeling and its application of the RPS23 genes as sheep immune character | |
| CN106755371A (en) | Method and its application using PCR RFLP detection sheep PCNP gene mononucleotide polymorphisms | |
| CN111057771B (en) | SNP molecular marker for distinguishing 'Zhongyang No. 1' from common fugu obscurus and application thereof | |
| CN102605064A (en) | Multi-gene pyramiding breeding method for thoroughbred milk goats | |
| CN113774149B (en) | Molecular marker of neoseiulus barkeri for resistance to chlorpyrifos and application and detection method thereof | |
| CN109055386A (en) | Silkworm BmSCP1 gene and its recombinant expression carrier and application | |
| CN106755474A (en) | The detection method of one herd boar KDM5B gene insertion/deletions and its application | |
| CN100460506C (en) | Omega-3-aliphatic acid desaturase, and its encoding gene and use thereof | |
| CN110592122B (en) | Zebra fish naalad2 gene promoter and application thereof | |
| CN102618528A (en) | Deleting system for long fragments in genome based on TALEN (transcription activator-like effector Nuclease) and ssODNs (single stranded oligonucleotides) and application thereof | |
| CN101845506A (en) | Breeding method of multi-gene pyramiding of fine breed milk goats | |
| CN105385764B (en) | Molecular labeling and its application of the STMN2 gene as sheep immune character |
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 | ||
| ASS | Succession or assignment of patent right |
Owner name: BEIJING GAILANDE INVESTMENT AND DEVELOPMENT CENTER Free format text: FORMER OWNER: INST. OF ANIMAL HUSBANDRY, CHINESE ACADEMY OF AGRICULTURAL SCIENCES Effective date: 20150715 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20150715 Address after: 100000 Beijing city Dongcheng District Dongzhimen South Street No. 5 CYTS Building Room 601 Patentee after: Beijing Gaillaud Investment Development Center Address before: 100094 Animal Husbandry Institute, Chinese Academy of Agricultural Sciences, 2 West Old Summer Palace Road, Beijing, Haidian District Patentee before: INST ANIMAL SCIENCES CAAS |
|
| CI01 | Publication of corrected invention patent application |
Correction item: Patentee|Address Correct: Beijing Gaillaud Investment Development Centre (limited partnership)|Beijing city Dongcheng District Dongzhimen South Street No. 5 CYTS Building Room 601 False: Beijing Gaillaud Investment Development Center|Beijing city Dongcheng District Dongzhimen South Street No. 5 CYTS Building Room 601 Number: 23 Volume: 20 |
|
| ERR | Gazette correction |
Free format text: CORRECT: PATENTEE; ADDRESS; FROM: BEIJING GAILANDE INVESTMENT AND DEVELOPMENT CENTER; DONGCHENG, BEIJING TO: DONGCHENG, BEIJING; DONGCHENG, BEIJING |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20151013 Address after: Beijing City, Dongcheng District Dongzhimen CYTS 100007 building 6 layer 601 Patentee after: GRAND LIFE SCIENCE & TECHNOLOGY, Ltd. Address before: Beijing city Dongcheng District Dongzhimen South Street No. 5 CYTS Building Room 601 Patentee before: Beijing Gaillaud Investment Development Centre (L.P.) |
|
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20060503 |