马氏珠母贝生长与生物矿化相关基因的SNP标记开发
|
摘要
马氏珠母贝(Pinetada martensii Dunker)是我国主要的海水珍珠生产贝类。提高育珠贝的生长性状(如壳高,壳宽和壳厚、壳重和总重)和珍珠质分泌性状(如珍珠层厚度、珍珠质量等)是马氏珠母贝遗传改良的首要目标。而筛选与生长以及珍珠质分泌性状显著关联的分子标记,用于辅助选择,是马氏珠母贝生长和育珠性状进行有效选择的高效而可靠的手段与途径。在研究策略上,利用畜禽等农业动物育种中功能强大的筛选分析候选基因的“整合信息学策略”原理,系统分析我们建立和获得的马氏珠母贝ESTs序列。筛选与生长性状关联的、潜在的候选QTL Gene以及己知生物矿化基因,利用扩增重测序技术,检测其上的单核苷酸多态性(SNP)位点,开发SNP标记;并在马氏珠母贝野生群体和家系中检测其遗传特性和遗传模式,进而在分离群体中检测与生长和珍珠质分泌性状相关的SNP标记,达到辅助选育的目的。此外由于其在基因组中分布的丰富性和同一位点双等位基因的优点可使遗传作图更加精细。基于SNP的连锁图谱不仅增加了标记密度,提供与丰富基因联系的物理图谱,从而有利于遗传图谱和物理图谱的整合;而且为数量性状基因座和定位候选基因鉴定提供了精细图。
本研究对马氏珠母贝的EST数据库进行生物信息学分析,获得与生长性状关联的、潜在的候选QTL Gene或片段,以及已知生物矿化基因序列；据此设计引物174对。在马氏珠母贝6个不同地理群体的12个个体中进行扩增和扩增产物重测序,align分析测得的序列,在总计33Kb的马氏珠母贝测序基因组中,通过分析推断出354个SNP位点,平均每94bp发现一个SNP位点。在所有位点中选择40个SNP位点,针对每个SNP位点设计前后端引物,其中27对引物扩增后在聚丙烯酰胺凝胶上呈现单一条带。为这27个SNP位点设计目标SNP位点居中,长度为20-35bp的探针。PCR反应后,用非标记探针高分辨率溶解曲线(HRM)的方法检测SNP的基因型。在三个马氏珠母贝野生群体(popl：2003年采自三亚亚龙湾,pop2：2009年采自三亚安游,pop3：2010年采自三亚南山,每个群体n=48)和一个家系(n=48)中,17个SNP标记获得稳定的基因型数据。其中一个标记为单态,其余16个标记呈多态。16个多态位点中,位点act在三个野生群体中都显著偏离HWE,位点pris-2在第一和第二个群体中显著偏离HWE,位点mec-7a在第二、三个群体显著偏离HWE,位点hsp-1在第一个群体中显著偏离HWE(P<0.05并经Bonferroni多重比较校正)。其中位点act和mec-7a在偏离的群体中均呈现显著地杂合子过剩(P<0.01)。而位点act和pris-2在偏离的群体中均呈现显著的杂合子缺失(P<0.01)。这些位点可能主要受到选择压力。16个位点在3个野生群体的绝大部分个体中都能正常扩增,且三个群体的整体平均Fst仅为0.0048。说明16个位点上三个群体间的遗传差异不显著。
16个多态位点中除两个位点(mec-7b和ben-1)外,在马氏珠母贝家系中均有分离。14个分离中,ddx-23和hsp-1两个位点显著偏离孟德尔遗传分离比(P<0.05)。显示这两个位点受到选择压力。鉴于所有的SNP位点来源于与生长发育基因相关的ESTs序列和贝类生物矿化以及珍珠质蛋白基因,对这些位点进一步的与相关性状的关联分析以及在遗传连锁图谱上的定位和QTL分析都将揭示它们所在基因功能及其作用模式。为马氏珠母贝分子标记辅助育种以及功能基因的相关研究奠定基础。
Pearl oyster Pinctada martensii (Dunker) is the main species for marine pearl production in China.Pearl oyster growth traits (such as shell height, shell width and shell thickness, shell weight and total weight) and the secretion of nacre traits (such as nacre thickness, pearl quality, etc.) is the primary goal of genetic improvement of Pinctada martensii. However Screening with the growth and nacre secretion traits are significantly associated molecular markers for assisted selection, as Pinctada martensii growth and fertility beads traits for effective choice of efficient and reliable means and Approaches. Integrating informatics strategies " is a method of screening analysis of candidate genes in animal breeding of poultry and other agricultural, that systematic analysis Pinctada martensii ESTs of we establishment and gained. In this study, screening significant effects on growth、potential candidate genes for QTL and known Biomineralization genes. Use of amplified re-sequencing Technology to detect and Development single nucleotide polymorphism (SNP) loci. Detection of genetic characteristics and genetic model in the wild populations and family of Pinctada martensii, and then Detection of SNP markers associated with the growth and nacre secretion traits in the Segregating populations. In addition, due to the advantages of Richness in the genome distribution and the same Sites Biallelic make more refined in genetic mapping. SNP-based linkage map not only increased the marker density, provides the physical map with the rich genetic link, thus contributing to the integration of genetic maps and physical maps; and provides a detailed map of quantitative trait loci and QTL gene identification.
In this study, bioinformatics analysis of the EST database of Pinctada martensii, get the significant effects on growth、potential candidate genes for QTL and known Biomineralization genes. Primers were designed for174, Amplification and amplification products re-sequenced in12individuals of six different geographical populations in Pinctada martensii. Use of Align analysis of the measured, three hundred and fifty-four putative SNPs were discovered by resequencing33Kb, corresponding to a very high density of one SNP per94bp. Select the40SNPs in all sites, Primers were designed for each SNPs,27pairs of primers showed a single band on polyacrylamide gel. Design length of20-35bp probe for the27SNPs, After PCR, the non-labeled probe High Resolution Melting (HRM) to detect the SNP genotype. In three wild populations(popl:Collected from Sanya Yalong Bay in2003、pop2: Collected from Sanya anyou in2009pop3:Collected from Sanya Nanshan in2010) and a family (48individuals) of Pinctada martensii, the17SNP markers to obtain a stable genotype data. The one marked as a single state, the remaining16markers were polymorphic. of which act showed significant (P <0.05after Bonferroni correction) departures from Hardy-einberg equilibrium in all the populations with heterozygote excess,while pris-2,mec-7a and hsp-1deviated from HWE in at least one population. act and mec-7a in the deviation from the group showed a significantly to heterozygotes excess (P <0.01), act and pris-2in the deviation from the group showed a significant to deficiency (P <0.01). These sites may be subject to selection pressure.16SNPs can be amplified in the vast majority of individuals in three wild populations, the overall average Fst was only0.0048. Description of the16genetic differences was not significant between the three groups
Fourteen (excluding mec-7b and ben-1) of the16validated SNPs segregated in a mapping family, ddx-23and hsp-1significantly deviated from the Mendelian ratios(P<0.05), Show these two sites subject to selection pressure. View of all SNP loci derived from ESTs sequences that related to the growth of gene and shellfish biomineralization of nacre protein gene, these sites Related Traits in analysis and position on the genetic linkage map and QTL analysis will reveal where the gene function and its mode of action. Lay the foundation for the Pinctada martensii molecular marker assisted breeding and functional genes.
本研究对马氏珠母贝的EST数据库进行生物信息学分析,获得与生长性状关联的、潜在的候选QTL Gene或片段,以及已知生物矿化基因序列；据此设计引物174对。在马氏珠母贝6个不同地理群体的12个个体中进行扩增和扩增产物重测序,align分析测得的序列,在总计33Kb的马氏珠母贝测序基因组中,通过分析推断出354个SNP位点,平均每94bp发现一个SNP位点。在所有位点中选择40个SNP位点,针对每个SNP位点设计前后端引物,其中27对引物扩增后在聚丙烯酰胺凝胶上呈现单一条带。为这27个SNP位点设计目标SNP位点居中,长度为20-35bp的探针。PCR反应后,用非标记探针高分辨率溶解曲线(HRM)的方法检测SNP的基因型。在三个马氏珠母贝野生群体(popl：2003年采自三亚亚龙湾,pop2：2009年采自三亚安游,pop3：2010年采自三亚南山,每个群体n=48)和一个家系(n=48)中,17个SNP标记获得稳定的基因型数据。其中一个标记为单态,其余16个标记呈多态。16个多态位点中,位点act在三个野生群体中都显著偏离HWE,位点pris-2在第一和第二个群体中显著偏离HWE,位点mec-7a在第二、三个群体显著偏离HWE,位点hsp-1在第一个群体中显著偏离HWE(P<0.05并经Bonferroni多重比较校正)。其中位点act和mec-7a在偏离的群体中均呈现显著地杂合子过剩(P<0.01)。而位点act和pris-2在偏离的群体中均呈现显著的杂合子缺失(P<0.01)。这些位点可能主要受到选择压力。16个位点在3个野生群体的绝大部分个体中都能正常扩增,且三个群体的整体平均Fst仅为0.0048。说明16个位点上三个群体间的遗传差异不显著。
16个多态位点中除两个位点(mec-7b和ben-1)外,在马氏珠母贝家系中均有分离。14个分离中,ddx-23和hsp-1两个位点显著偏离孟德尔遗传分离比(P<0.05)。显示这两个位点受到选择压力。鉴于所有的SNP位点来源于与生长发育基因相关的ESTs序列和贝类生物矿化以及珍珠质蛋白基因,对这些位点进一步的与相关性状的关联分析以及在遗传连锁图谱上的定位和QTL分析都将揭示它们所在基因功能及其作用模式。为马氏珠母贝分子标记辅助育种以及功能基因的相关研究奠定基础。
Pearl oyster Pinctada martensii (Dunker) is the main species for marine pearl production in China.Pearl oyster growth traits (such as shell height, shell width and shell thickness, shell weight and total weight) and the secretion of nacre traits (such as nacre thickness, pearl quality, etc.) is the primary goal of genetic improvement of Pinctada martensii. However Screening with the growth and nacre secretion traits are significantly associated molecular markers for assisted selection, as Pinctada martensii growth and fertility beads traits for effective choice of efficient and reliable means and Approaches. Integrating informatics strategies " is a method of screening analysis of candidate genes in animal breeding of poultry and other agricultural, that systematic analysis Pinctada martensii ESTs of we establishment and gained. In this study, screening significant effects on growth、potential candidate genes for QTL and known Biomineralization genes. Use of amplified re-sequencing Technology to detect and Development single nucleotide polymorphism (SNP) loci. Detection of genetic characteristics and genetic model in the wild populations and family of Pinctada martensii, and then Detection of SNP markers associated with the growth and nacre secretion traits in the Segregating populations. In addition, due to the advantages of Richness in the genome distribution and the same Sites Biallelic make more refined in genetic mapping. SNP-based linkage map not only increased the marker density, provides the physical map with the rich genetic link, thus contributing to the integration of genetic maps and physical maps; and provides a detailed map of quantitative trait loci and QTL gene identification.
In this study, bioinformatics analysis of the EST database of Pinctada martensii, get the significant effects on growth、potential candidate genes for QTL and known Biomineralization genes. Primers were designed for174, Amplification and amplification products re-sequenced in12individuals of six different geographical populations in Pinctada martensii. Use of Align analysis of the measured, three hundred and fifty-four putative SNPs were discovered by resequencing33Kb, corresponding to a very high density of one SNP per94bp. Select the40SNPs in all sites, Primers were designed for each SNPs,27pairs of primers showed a single band on polyacrylamide gel. Design length of20-35bp probe for the27SNPs, After PCR, the non-labeled probe High Resolution Melting (HRM) to detect the SNP genotype. In three wild populations(popl:Collected from Sanya Yalong Bay in2003、pop2: Collected from Sanya anyou in2009pop3:Collected from Sanya Nanshan in2010) and a family (48individuals) of Pinctada martensii, the17SNP markers to obtain a stable genotype data. The one marked as a single state, the remaining16markers were polymorphic. of which act showed significant (P <0.05after Bonferroni correction) departures from Hardy-einberg equilibrium in all the populations with heterozygote excess,while pris-2,mec-7a and hsp-1deviated from HWE in at least one population. act and mec-7a in the deviation from the group showed a significantly to heterozygotes excess (P <0.01), act and pris-2in the deviation from the group showed a significant to deficiency (P <0.01). These sites may be subject to selection pressure.16SNPs can be amplified in the vast majority of individuals in three wild populations, the overall average Fst was only0.0048. Description of the16genetic differences was not significant between the three groups
Fourteen (excluding mec-7b and ben-1) of the16validated SNPs segregated in a mapping family, ddx-23and hsp-1significantly deviated from the Mendelian ratios(P<0.05), Show these two sites subject to selection pressure. View of all SNP loci derived from ESTs sequences that related to the growth of gene and shellfish biomineralization of nacre protein gene, these sites Related Traits in analysis and position on the genetic linkage map and QTL analysis will reveal where the gene function and its mode of action. Lay the foundation for the Pinctada martensii molecular marker assisted breeding and functional genes.
引文
[1]黎裕,贾继增,王天宇等.分子标记的种类及其发展[J].生物技术通报,1999(4)：19-22.
[2]管敏强,陈锡文等.分子标记技术及其应用[J].实验动物科学与管理,2005,22(1)：48-51.
[3]Botstein D R, White R L, Skolnick M, et al. Construction of a geneticlinkage map in man using restriction fragment length polymorphism[J]. Am J Hum Genet,1980,32:314-331
[4]Huang N.pyramiding of bacterial blight resistance genes in rice maker-aid selection using RFLP and PCR[J].Theor Appl. Genet,1997 (95):313-320
[5]张汉尧RFLP在植物遗传育种研究中的应用[J].西南林学院学报,2006,26(1)：75-80.
[6]周宇爝.分子标记发展简史[J].现代农业科技,2009,11：264-270
[7]Orbacz EA, Wilbur AE, Wakefield JR, Gaffney PM (1996) RFLP analysis of genetic diversity in a Siberian population of the Japanese scallop (Patinopecten yessoensis). Journal of Shellfish Research,15,529.
[8]Cordeiro,G.M.,Casu,R.Mclntyre,C.L.,Manners,J.M.,Henry,R.J.(2001) Microsatellite markers from sugareane(Saccharum spp.)ESTs cross transferable to erianthus and sorghum.Plant Scienee,160:1115-1123.
[9]Crooijmans,R.,Beirbooms,N.,KomenJ,et al.(1997) Microsatellite markers in common carp[J].Animal Geneties,28:129-132
[10]Zane L, Bargelloni L, Patarnello T. Strateg ies for m icrosatellites isolation:areview [J].Mol Ecol,2002,11:1-16.
[11]刘瑾,舒妙安.微卫星标记及其在贝类中的应用[J].水利渔业,2007,27(2)：17-19.
[12]李琪.海洋贝类微卫星DNA标记的开发及其在遗传学研究中的应用[J].中国水产科学,2006,13(3)：502-509.
[13]Zabean M, VosP.Selective reseriction fragment amplification:a general method for DNA fingerping. Euro pean patent Application 9240-2629[D],1993-03-31.
[14]王斌,翁曼丽AFLP的原理及其应用[J].杂交水稻,2003(5)：27-30.
[15]王晓涛,姚志刚,瓮巧云AFLP分子标记技术及其应用[J].农技服务,2010,27(8)：1099-11OO
[16]Wang D. G., Fan J. B., Siao C. J., et al. Large-scale identification, mapping and genotyping of single nucleotide polymorphisms in the human genome. Science,1998,280:1077-1082.
[17]Aumann Y, Man Isterski E, Yakh Ni Zi Designing op timally multip lexed SNP genotyping assays [J]. J Comput Syst Sci,2005,70 (3):399-4171
[18]Brumfield R T, Beerli P, Nickersond A, et all The utility of single nucleotide polymorphisms in inferences of population history [J]. Trends Ecol Evol,2003,18 (5):249-2561
[19]Herbergs J., Siwek M., Crooijmans R. P., et al.Multicolour fluorescent detection and mapping of AFLP markers in chicken (Gallus domesticus). Anim Genet,1999,30(4):274-285.
[20]Knorr C., Cheng H. H.,Dodgson J. B.. Application of AFLP markers to genome mapping in poultry. Anim Genet,1999,30(1):28-35.
[21]Vuylsteke M., Peleman J. D., van Eijk M. J. AFLP-based transcript profiling (cDNA-AFLP) for genome-wide expression analysis. Nat Protoc,2007,2(6):1399-413.
[22]Collins FS,Brooks LD, Chakravarti A.A DHA DNA polymorphism discovery resource for research on human genetic variation[J].Genome Res,1998,8:1229-1231.
[23]Wong G. K., Liu B., Wang J., et al. A genetic variation map for chicken with 2.8 million single nucleotide polymorphisms. Nature,2004,432(7018):717-722.
[24]Cui J. X., Du H. L.,Zhang X. Q. Rapidly Screening SNPs and Estimating Allelic Frequencies by DNA Pooling and Sequencing. Acta Genetica Sinica,2005,32(4):372-377.
[25]Mullikin J. C., Hunt S. E., Cole C. G., et al. An SNP map of human chromosome 22. Nature, 2000,407:516-520.
[26]Wang W. P., Ni K. Y., Zhou G. H. approaches for SNP genotyping, Hereditas (Beijing),2006, 28(1):117-126.
[27]Yoshinori Akagi, Megumi Makimura, Yoshiyuki Yokoyama, et al.Development of a ligation-based impedimetric DNA sensor for single-nucleotide polymorphism associated with metabolic syndrome[J]. Electrochimica Acta,2006,51:6367-6372
[28]Zhang R., Zhu Z., Zhu H., et al. SNP Cutter:a comprehensive tool for SNP PCR-RFLP assay design. Nucleic Acids Res,2005,33:489-492.
[29]Gu Y. J., Guo J. Y, Cheng H. M., et al. The Identification and Application of Single Nucleotide Polymorphisms. Chinese Agricultural Science Bulletin,2007,23(4):38-41.
[30]Morin PA, Saiz R, Monjazeb A (1999) High-throughput single nucleotide polymorphism genotyping by fluorescent 5'exonuclease assay. BioTechniques,27,538-552.
[31]范新兰.HLA-DRA基因进化及DRB可表达基因调控区SNPs研究[D].中山大学学位论文,2004
[32]Gu Y. J., Guo J. Y., Cheng H. M., et al. The Identification and Application of Single Nucleotide Polymorphisms. Chinese Agricultural Science Bulletin,2007,23(4):38-41.
[33]Amos C. I., Frazier M. L., Wang W. DNA pooling in mutation detection with reference to sequence analysis. Am J Hum Genet,2000,66(5):1689-92.
[34]Ou J. T., Tang S. Q., Sun D. X., et al. Polymorphisms of three neuroendocrine-correlated genes associated with growth and reproductive traits in the chicken. Poult Sci,2009,88(4): 722-7.
[35]Cui J. X., Du H. L.,Zhang X. Q. Rapidly Screening SNPs and Estimating Allelic Frequencies by DNA Pooling and Sequencing. Acta Genetica Sinica,2005,32(4):372-377.
[36]Ronaghi M.Genome Res.,2001,11(1):3-11
[37]周国华,古卓良,章杰兵.生物发光分析法检测P53基因上的突变点.药学学报,2002,37(1)：41-45
[38]Alderborn A, Kristofferson A, Hammerling U. Determination of single nucleotide polymorphisms by real2time pyrophosphate DNA sequencing. Genom e Res,2000,10: 1249-1258.
[39]Pourmand N, Elahi E, Davis RW, RonaghiM. Multip lex pyrosequencing. N ucleic Acids Res, 2002,30-31.
[40]张迟.生物标志物的检测方法研究[D].华中科技大学学位论文,2006
[41]Xiao W., Oefner P. J. Denaturing high-performance liquid chromatography:a review. Hum Mutat,2001,17(6):439-474
[42]Muyzer G, Smalla K.Application of denaturing gradient gelelectrophoresis (DGGE) and temperature gradient gelelectrophoresis (TGGE) in microbial ecology [J].Antoni van Leeuwenhoek,1998,73:127-141
[43]Orita M, Iwahana H, Kanazawa H, et al. Detection of polymorphismsof human DNA by gel electrophoresis as single-strand conformation polymorphisms[J]. Proc Natl Acad Sci USA, 1989,86:2766-2770.
[44]Gasser R. B., Hu M., Chilton N. B., et al. Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation. Nature Protocols,2006,1(6):3121-3128.
[45]Kakavas V. K., Plageras P., Vlachos T. A., et al. PCR-SSCP:a method for the molecular analysis of genetic diseases. Mol Biotechnol,2008,38(2):155-63.
[46]Murakami Y, Katahira M, Makino R, et al. Inactivation of the retinoblastoma gene in a human lung carcinoma cell line detected by single-strand conformation polymorphism analysis of the polymerase chain reaction product of cDNA[J]. Oncogene,1991,6:37-42.
[47]Risesner D, Steger G, Wiese U, et al. Temperature-gradientgel electrophoresis for the detection of polymorphic DNA and for quantitative polymerase chain reaction[J]. Electrophoresis,1992,13(9):63-66.
[48]Ota M., Fukushima H., Kulski J. K., et al. Single nucleotide polymorphism detection by polymerase chain reaction-restriction fragment length polymorphism. Nature Protocols,2007, 2(11):2857-2864.
[49]Ke X., Collins A., Ye S. PIRA PCR designer for restriction analysis of single nucleotide polymorphisms. Bioinformatics,2001,17(9):838-9
[50]Del Tito B J, Poff H E, Novotny M A, et al. Automated fluorescentanalysis procedure for enzymatic mutationdetection[J].Clinic Chem,1998,44:731-739.
[51]Chen, X. and P.F. Sullivan.2003. Single nucleotide polymorphism genotyping:biochemistry, protocol, cost and throughput. Pharmacogenomics J.3:77-96.
[52]Hardenbol, P., J. Baner, M. Jain, M. Nilsson, E.A. Namsaraev, G.A. Karlin-Neumann, H. Fakhrai-Rad, M. Ronaghi, et al.2003. Multiplexed genotyping with sequence-tagged molecular inversion probes. Nat. Biotechnol.21:673-678.
[53]Medintz, I., W.W. Wong, L. Berti, L. Shiow, J. Tom, J. Scherer, G. Sensabaugh, and R.A. Mathies.2001. High-performance multiplex SNP analysis of three hemochromatosis-related mutations with capillary array electrophoresis microplates. Genome Res.11:413-421.
[54]Jun Wang, Karen Chuang, et al.2005.High-throughput SNP genotyping by single-tube PCR with Tm-shift primers.BioTechniques 39:885-893
[55]Wang J,Chuang K, Ahluwalia M,Patel S,Umblas N,Mirel D,Higuchi R,and Germer S(2005) High-throughput SNP genotyping by single-tube PCR with Tm-shift primers,BioTechniques,39:885-893
[56]SHEN GQ, LUO A, WANGQ K1 High-throughput single-nucleotide polymorphisms genotyping:TaqMan assay and pyrosequencing assay [J]. MethodsMolMed,2006,128:209-2241
[57]Ranadek, Changms, Tingct, et all High-throughput genotyp ing with single nucleotide polymorphisms [J]. Genome Res,2001,11 (7):1262-12681
[58]Margrafr L, Mao R, Highsmith E, et al. Mutation scanning of the RET protooncogene using high-resolution melting analysis[J]. Clin Chem,2006,52:138-141.
[59]Slinger R, Bellfoy D, Desjardins M, et al. Highresolution melting assay for the detection of gyrA mutations causing quinolone resistance in Salmonella enteric serovars Typhi and Paratyphi[J].Diagn Microbiol Infect Dis,2007,57:455-458.
[60]Wittwerc T,Reed H,Gundrtc N, et al. Highresolution genotyping by amplicon melting analysis using LC Green[J]. Clin Chem,2003,49:853-860.
[61]Choul S, Lyon E, Wittwercl T. A comparison of highresolution melting analysis with denaturing high-performance liquid chromatography for mutation scanning cystic fibrosis transmembrane conductance regulator gene as a model[J]. Am J Clin Pathol,2005,124: 330-338.
[62]ChengJ C, Huang CL, Lin CC, et al. Rapid detection and identification of clinically important bacteria by highresolution melting analysis after broad-range ribosomal RNA real-time PCR[J]. Clin Chem,2004,52:1997-2004.
[63]Zhou L,Vandersteen J,WangL1Tissue Antigens,2004,64:156～1641
[64]Wang D G, Fan J B, Siao C J, et al. Large-scale identification, mapping, and genotyping of single-nucleotide polymorphism in the human genome[J]. Science,1998,280:1077-1 082.
[65]Zhu J., Zhang B., Smith E. N., et al. Integrating large-scale functional genomic data to dissect the complexity of yeast regulatory networks. Nat Genet,2008,40 (7):854-61
[66]Pletcher, M.T., McClurg, P., Batalov, S., Su, A.I., Barnes, S.W., Lagler, E., Korstanje, R.,Wang, X., Nusskern, D., Bogue, M.A., Mural, R.J., Paigen, B., Wiltshire, T.,2004. Use ofa dense single nucleotide polymorphism map for in silico mapping in the mouse.PLoS Biol.2, e393.
[67]Kucuktas H, Wang S L, Li P, et all Construction of geneticlinkage map s and comparative genome analysis of catfish usinggene-associated markers [J]. Genetics,2009,181 (4):1649-16601
[68]Zhang, L., Guo, X., Development and validation of single nucleotide polymorphism markers in the eastern oyster Crassostrea virginica Gmelin by mining ESTs and resequencing, Aquaculture (2010), doi:10.1016/j.aquaculture.2010.02.012
[69]Hayesb, Laerdahlj K, Lien S, et all An extensive resourceof single nucleotide polymorphism markers associated with Atlanticsalmon (Salmo salar) exp ressed sequences [J]. Aquac, 2007,265 (1/4):82-91
[70]Hutt F. B. Genetics of the fowl. VI. A tentative chromosome map. Neue Forschung fur Tierzuchtund Abstammung,1936:105-112.
[71]Keller B., Feuillet C., Yahiaoui N. Map-based isolation of disease resistance genes from bread wheat:cloning in a supersize genome. Genet Res,2005,85(2):93-100.
[72]Levin 1., Santangelo L., Cheng H., et al. An autosomal genetic linkage map of the chicken. J Hered,1994,85(2):79-85.
[73]Sauer S., Reinhardt R., Lehrach H., et al. Single-nucleotide polymorphisms:analysis by mass spectrometry. Nature Protocols,2006,1(4):1761-1771.
[74]Lee, J.H., Guo, X.,2006. Mining EST database for single-nucleotide polymorphisms in the eastern oyster (Crassostrea virginica). J. Shellfish Res.25,748-749. Abstract.
[75]He F, Wenh S, Dongs L, et all Identification of estrogen recep tor alpha gene polymorphisms by SSCP and its effect on rep roductive traits in Japanese flounder (Paralichthys olivaceus) [J].Comp Biochem Physiol B:Biochem Mol Biol,2008,150 (3):278-2831
[76]Rengmark A H, L ingaas F Genomic structure of the Nile tilap ia (O reochrom is niloticus) transferrin gene and a hap lotype associated with saltwater tolerance [J]. Aquac,2007,272 (1/4):146-1551
[77]Tatsuda K., Fujinaka K. Genetic mapping of the QTL affecting body weight in chickens using a F2 family. Br Poult Sci,2001,42(3):333-337.
[78]Wilhelm B. T., Marguerat S., Watt S., et al. Dynamic repertoire of a eukaryotic transcriptome surveyed at single-nucleotide resolution. Nature,2008,453(7199):1239-43.
[79]Elfstromc M, Gaffneyp M, Smithc T, et all Characterization of 12 single nucleotide polymorphisms in weathervane scallop[J]. Mol Ecol Notes,2005,5 (2):406-4091
[80]Aranishi, E., Okimoto, T. and Izumi, S. (2005 a) Identification of gadoid species (Pisces, Gadidae) by PCR-RFLP analysis. J. Appl. Genet.,46,69-73.
[81]Smith C T, Templin W D, Seeb J E, et al. Single nucleotide polymorphisms provide rapid and accurate estimates of the proportions of U.S. and Canadian Chinook salmon caught in Yukon River fisheries[J]. North American J Fish Manag,2005,25:944-953.
[82]Itoi S, Nakaya M, Kaneko G, et al. Rapid identification of eels Anguilla japonica and Anguilla anguilla by polymerase chain reaction with single nucleotide polymorphism-based specific probes[J]. Fish Sci,2005,71:1356-1364.
[83]Ye X., McLeod S., D. Elfick, Dekkers J. C., et al. Rapid identification of single nucleotide polymorphisms and estimation of allele frequencies using sequence traces from DNA pools. Poult Sci,2006,85:1165-1168.
[84]王爱民,石耀华.2003.中国马氏珠母贝遗传育种的现状与展望.农业生物技术学报,11(6)：547-553
[85]顾志峰,王嫣,石耀华等.2009.马氏珠母贝2个不同地理种群的形态性状和贝壳珍珠质颜色比较分析.渔业科学进展,30：79-86
[86]Beyer A., Bandyopadhyay S., Ideker T. Integrating physical and genetic maps:from genomes to interaction networks. Nat Rev Genet,2007,8(9):699-710.
[87]Ge H., Walhout A. J., Vidal M. Integrating 'omic' information:a bridge between genomics and systems biology. Trends Genet,2003,19(10):551-60.
[88]Germer S, Higuchi R (2003) Homogeneous allele-specific PCR in SNP genotyping. Methods in Molecular Biology,212,197-214.
[89]陈炜,张戈,张思仲.基于生物信息学的SNP候选位点搜寻方法[J].遗传,2001,23(2)：153-156
[90]Frazerk A, Ballingerd G, Coxd R, et al. A second generation human haplotype map of over3. 1 million SNPs[J]. Nature,2007,449:851-861
[91]Tenaillon M, Sawkinsmc, Longa D,et al. Patterns of DNA sequence polymorphism a 1 ong chromosome of maize (Zea mays ssp.maysL.)[J].ProcNatlAcad US A Sci,2001,9 8(16):9161-9166
[92]Sauvagec, Bieren, Lapegues, et al.Single nucleotide polymorphisms and their relationship to codonusage bias in the Pacific oyter Crassostreag igas [J].Gene,2007,406(1-2):13-22
[93]Lirh, Li Q, Konglf. Characterization of expressed sequence tag-derived single-nucleotide polymorphisms in the bay scallop Argopecten irradians [J]. Fish Sci 2009,75(6):1389-1400
[94]Zhang, L., Guo, X., Development and validation of single nucleotide polymorphism markers in the eastern oyster Crassostrea virginica Gmelin by mining ESTs and resequencing, Aquaculture (2010), doi:10.1016/j.aquaculture.2010.02.012
[95]吕敏芝2007.MS Excel在畜牧兽医统计中的应用(四)—卡方检验(X2检验).广东畜牧兽医科技,32(2)：30-32
[96]Richardi,Beckman J S. How neutral are synonymous codon mutations[J]. N at Genet, 1995,10:259.
[97]Carglll,M.,Altshuler,D.,Ireland,J.,Sklar,p.,Ardlie,K.Patil,N.,Shaw,N.,Lane,CR.,Lim.EP.,Kalya naraman,N.(1999)Charaeterization of single-nucleotide Polymorphisms in coding regions of human genes.NatGenet,22:231-38
[98]Nielsen,R.(2005)Molecular signatures of natural seleetion.Annu Rev Genet,39:197-218
[99]Francesca Capon,Richard C.Trembath,et al.(2004)A synonymous SNP of the corneodesmosin gene leads to increased mRNA stability and demonstrates association with psoriasis across diverse ethnic groups.Hum. Mol. Genet.13 (20):2361-2368.
[100]Giancarlo V. De Ferrari,et al.(2007)Common genetic variation within the Low-Density Lipoprotein Receptor-Related Protein 6 and late-onset Alzheimer's disease.PNAS May 29,vol.104 no.22:9434-9439
[101]Komar,A(2007) SNPs,Slient But Not Invisible,Science,315(5811),466-467(2007)
[2]管敏强,陈锡文等.分子标记技术及其应用[J].实验动物科学与管理,2005,22(1)：48-51.
[3]Botstein D R, White R L, Skolnick M, et al. Construction of a geneticlinkage map in man using restriction fragment length polymorphism[J]. Am J Hum Genet,1980,32:314-331
[4]Huang N.pyramiding of bacterial blight resistance genes in rice maker-aid selection using RFLP and PCR[J].Theor Appl. Genet,1997 (95):313-320
[5]张汉尧RFLP在植物遗传育种研究中的应用[J].西南林学院学报,2006,26(1)：75-80.
[6]周宇爝.分子标记发展简史[J].现代农业科技,2009,11：264-270
[7]Orbacz EA, Wilbur AE, Wakefield JR, Gaffney PM (1996) RFLP analysis of genetic diversity in a Siberian population of the Japanese scallop (Patinopecten yessoensis). Journal of Shellfish Research,15,529.
[8]Cordeiro,G.M.,Casu,R.Mclntyre,C.L.,Manners,J.M.,Henry,R.J.(2001) Microsatellite markers from sugareane(Saccharum spp.)ESTs cross transferable to erianthus and sorghum.Plant Scienee,160:1115-1123.
[9]Crooijmans,R.,Beirbooms,N.,KomenJ,et al.(1997) Microsatellite markers in common carp[J].Animal Geneties,28:129-132
[10]Zane L, Bargelloni L, Patarnello T. Strateg ies for m icrosatellites isolation:areview [J].Mol Ecol,2002,11:1-16.
[11]刘瑾,舒妙安.微卫星标记及其在贝类中的应用[J].水利渔业,2007,27(2)：17-19.
[12]李琪.海洋贝类微卫星DNA标记的开发及其在遗传学研究中的应用[J].中国水产科学,2006,13(3)：502-509.
[13]Zabean M, VosP.Selective reseriction fragment amplification:a general method for DNA fingerping. Euro pean patent Application 9240-2629[D],1993-03-31.
[14]王斌,翁曼丽AFLP的原理及其应用[J].杂交水稻,2003(5)：27-30.
[15]王晓涛,姚志刚,瓮巧云AFLP分子标记技术及其应用[J].农技服务,2010,27(8)：1099-11OO
[16]Wang D. G., Fan J. B., Siao C. J., et al. Large-scale identification, mapping and genotyping of single nucleotide polymorphisms in the human genome. Science,1998,280:1077-1082.
[17]Aumann Y, Man Isterski E, Yakh Ni Zi Designing op timally multip lexed SNP genotyping assays [J]. J Comput Syst Sci,2005,70 (3):399-4171
[18]Brumfield R T, Beerli P, Nickersond A, et all The utility of single nucleotide polymorphisms in inferences of population history [J]. Trends Ecol Evol,2003,18 (5):249-2561
[19]Herbergs J., Siwek M., Crooijmans R. P., et al.Multicolour fluorescent detection and mapping of AFLP markers in chicken (Gallus domesticus). Anim Genet,1999,30(4):274-285.
[20]Knorr C., Cheng H. H.,Dodgson J. B.. Application of AFLP markers to genome mapping in poultry. Anim Genet,1999,30(1):28-35.
[21]Vuylsteke M., Peleman J. D., van Eijk M. J. AFLP-based transcript profiling (cDNA-AFLP) for genome-wide expression analysis. Nat Protoc,2007,2(6):1399-413.
[22]Collins FS,Brooks LD, Chakravarti A.A DHA DNA polymorphism discovery resource for research on human genetic variation[J].Genome Res,1998,8:1229-1231.
[23]Wong G. K., Liu B., Wang J., et al. A genetic variation map for chicken with 2.8 million single nucleotide polymorphisms. Nature,2004,432(7018):717-722.
[24]Cui J. X., Du H. L.,Zhang X. Q. Rapidly Screening SNPs and Estimating Allelic Frequencies by DNA Pooling and Sequencing. Acta Genetica Sinica,2005,32(4):372-377.
[25]Mullikin J. C., Hunt S. E., Cole C. G., et al. An SNP map of human chromosome 22. Nature, 2000,407:516-520.
[26]Wang W. P., Ni K. Y., Zhou G. H. approaches for SNP genotyping, Hereditas (Beijing),2006, 28(1):117-126.
[27]Yoshinori Akagi, Megumi Makimura, Yoshiyuki Yokoyama, et al.Development of a ligation-based impedimetric DNA sensor for single-nucleotide polymorphism associated with metabolic syndrome[J]. Electrochimica Acta,2006,51:6367-6372
[28]Zhang R., Zhu Z., Zhu H., et al. SNP Cutter:a comprehensive tool for SNP PCR-RFLP assay design. Nucleic Acids Res,2005,33:489-492.
[29]Gu Y. J., Guo J. Y, Cheng H. M., et al. The Identification and Application of Single Nucleotide Polymorphisms. Chinese Agricultural Science Bulletin,2007,23(4):38-41.
[30]Morin PA, Saiz R, Monjazeb A (1999) High-throughput single nucleotide polymorphism genotyping by fluorescent 5'exonuclease assay. BioTechniques,27,538-552.
[31]范新兰.HLA-DRA基因进化及DRB可表达基因调控区SNPs研究[D].中山大学学位论文,2004
[32]Gu Y. J., Guo J. Y., Cheng H. M., et al. The Identification and Application of Single Nucleotide Polymorphisms. Chinese Agricultural Science Bulletin,2007,23(4):38-41.
[33]Amos C. I., Frazier M. L., Wang W. DNA pooling in mutation detection with reference to sequence analysis. Am J Hum Genet,2000,66(5):1689-92.
[34]Ou J. T., Tang S. Q., Sun D. X., et al. Polymorphisms of three neuroendocrine-correlated genes associated with growth and reproductive traits in the chicken. Poult Sci,2009,88(4): 722-7.
[35]Cui J. X., Du H. L.,Zhang X. Q. Rapidly Screening SNPs and Estimating Allelic Frequencies by DNA Pooling and Sequencing. Acta Genetica Sinica,2005,32(4):372-377.
[36]Ronaghi M.Genome Res.,2001,11(1):3-11
[37]周国华,古卓良,章杰兵.生物发光分析法检测P53基因上的突变点.药学学报,2002,37(1)：41-45
[38]Alderborn A, Kristofferson A, Hammerling U. Determination of single nucleotide polymorphisms by real2time pyrophosphate DNA sequencing. Genom e Res,2000,10: 1249-1258.
[39]Pourmand N, Elahi E, Davis RW, RonaghiM. Multip lex pyrosequencing. N ucleic Acids Res, 2002,30-31.
[40]张迟.生物标志物的检测方法研究[D].华中科技大学学位论文,2006
[41]Xiao W., Oefner P. J. Denaturing high-performance liquid chromatography:a review. Hum Mutat,2001,17(6):439-474
[42]Muyzer G, Smalla K.Application of denaturing gradient gelelectrophoresis (DGGE) and temperature gradient gelelectrophoresis (TGGE) in microbial ecology [J].Antoni van Leeuwenhoek,1998,73:127-141
[43]Orita M, Iwahana H, Kanazawa H, et al. Detection of polymorphismsof human DNA by gel electrophoresis as single-strand conformation polymorphisms[J]. Proc Natl Acad Sci USA, 1989,86:2766-2770.
[44]Gasser R. B., Hu M., Chilton N. B., et al. Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation. Nature Protocols,2006,1(6):3121-3128.
[45]Kakavas V. K., Plageras P., Vlachos T. A., et al. PCR-SSCP:a method for the molecular analysis of genetic diseases. Mol Biotechnol,2008,38(2):155-63.
[46]Murakami Y, Katahira M, Makino R, et al. Inactivation of the retinoblastoma gene in a human lung carcinoma cell line detected by single-strand conformation polymorphism analysis of the polymerase chain reaction product of cDNA[J]. Oncogene,1991,6:37-42.
[47]Risesner D, Steger G, Wiese U, et al. Temperature-gradientgel electrophoresis for the detection of polymorphic DNA and for quantitative polymerase chain reaction[J]. Electrophoresis,1992,13(9):63-66.
[48]Ota M., Fukushima H., Kulski J. K., et al. Single nucleotide polymorphism detection by polymerase chain reaction-restriction fragment length polymorphism. Nature Protocols,2007, 2(11):2857-2864.
[49]Ke X., Collins A., Ye S. PIRA PCR designer for restriction analysis of single nucleotide polymorphisms. Bioinformatics,2001,17(9):838-9
[50]Del Tito B J, Poff H E, Novotny M A, et al. Automated fluorescentanalysis procedure for enzymatic mutationdetection[J].Clinic Chem,1998,44:731-739.
[51]Chen, X. and P.F. Sullivan.2003. Single nucleotide polymorphism genotyping:biochemistry, protocol, cost and throughput. Pharmacogenomics J.3:77-96.
[52]Hardenbol, P., J. Baner, M. Jain, M. Nilsson, E.A. Namsaraev, G.A. Karlin-Neumann, H. Fakhrai-Rad, M. Ronaghi, et al.2003. Multiplexed genotyping with sequence-tagged molecular inversion probes. Nat. Biotechnol.21:673-678.
[53]Medintz, I., W.W. Wong, L. Berti, L. Shiow, J. Tom, J. Scherer, G. Sensabaugh, and R.A. Mathies.2001. High-performance multiplex SNP analysis of three hemochromatosis-related mutations with capillary array electrophoresis microplates. Genome Res.11:413-421.
[54]Jun Wang, Karen Chuang, et al.2005.High-throughput SNP genotyping by single-tube PCR with Tm-shift primers.BioTechniques 39:885-893
[55]Wang J,Chuang K, Ahluwalia M,Patel S,Umblas N,Mirel D,Higuchi R,and Germer S(2005) High-throughput SNP genotyping by single-tube PCR with Tm-shift primers,BioTechniques,39:885-893
[56]SHEN GQ, LUO A, WANGQ K1 High-throughput single-nucleotide polymorphisms genotyping:TaqMan assay and pyrosequencing assay [J]. MethodsMolMed,2006,128:209-2241
[57]Ranadek, Changms, Tingct, et all High-throughput genotyp ing with single nucleotide polymorphisms [J]. Genome Res,2001,11 (7):1262-12681
[58]Margrafr L, Mao R, Highsmith E, et al. Mutation scanning of the RET protooncogene using high-resolution melting analysis[J]. Clin Chem,2006,52:138-141.
[59]Slinger R, Bellfoy D, Desjardins M, et al. Highresolution melting assay for the detection of gyrA mutations causing quinolone resistance in Salmonella enteric serovars Typhi and Paratyphi[J].Diagn Microbiol Infect Dis,2007,57:455-458.
[60]Wittwerc T,Reed H,Gundrtc N, et al. Highresolution genotyping by amplicon melting analysis using LC Green[J]. Clin Chem,2003,49:853-860.
[61]Choul S, Lyon E, Wittwercl T. A comparison of highresolution melting analysis with denaturing high-performance liquid chromatography for mutation scanning cystic fibrosis transmembrane conductance regulator gene as a model[J]. Am J Clin Pathol,2005,124: 330-338.
[62]ChengJ C, Huang CL, Lin CC, et al. Rapid detection and identification of clinically important bacteria by highresolution melting analysis after broad-range ribosomal RNA real-time PCR[J]. Clin Chem,2004,52:1997-2004.
[63]Zhou L,Vandersteen J,WangL1Tissue Antigens,2004,64:156～1641
[64]Wang D G, Fan J B, Siao C J, et al. Large-scale identification, mapping, and genotyping of single-nucleotide polymorphism in the human genome[J]. Science,1998,280:1077-1 082.
[65]Zhu J., Zhang B., Smith E. N., et al. Integrating large-scale functional genomic data to dissect the complexity of yeast regulatory networks. Nat Genet,2008,40 (7):854-61
[66]Pletcher, M.T., McClurg, P., Batalov, S., Su, A.I., Barnes, S.W., Lagler, E., Korstanje, R.,Wang, X., Nusskern, D., Bogue, M.A., Mural, R.J., Paigen, B., Wiltshire, T.,2004. Use ofa dense single nucleotide polymorphism map for in silico mapping in the mouse.PLoS Biol.2, e393.
[67]Kucuktas H, Wang S L, Li P, et all Construction of geneticlinkage map s and comparative genome analysis of catfish usinggene-associated markers [J]. Genetics,2009,181 (4):1649-16601
[68]Zhang, L., Guo, X., Development and validation of single nucleotide polymorphism markers in the eastern oyster Crassostrea virginica Gmelin by mining ESTs and resequencing, Aquaculture (2010), doi:10.1016/j.aquaculture.2010.02.012
[69]Hayesb, Laerdahlj K, Lien S, et all An extensive resourceof single nucleotide polymorphism markers associated with Atlanticsalmon (Salmo salar) exp ressed sequences [J]. Aquac, 2007,265 (1/4):82-91
[70]Hutt F. B. Genetics of the fowl. VI. A tentative chromosome map. Neue Forschung fur Tierzuchtund Abstammung,1936:105-112.
[71]Keller B., Feuillet C., Yahiaoui N. Map-based isolation of disease resistance genes from bread wheat:cloning in a supersize genome. Genet Res,2005,85(2):93-100.
[72]Levin 1., Santangelo L., Cheng H., et al. An autosomal genetic linkage map of the chicken. J Hered,1994,85(2):79-85.
[73]Sauer S., Reinhardt R., Lehrach H., et al. Single-nucleotide polymorphisms:analysis by mass spectrometry. Nature Protocols,2006,1(4):1761-1771.
[74]Lee, J.H., Guo, X.,2006. Mining EST database for single-nucleotide polymorphisms in the eastern oyster (Crassostrea virginica). J. Shellfish Res.25,748-749. Abstract.
[75]He F, Wenh S, Dongs L, et all Identification of estrogen recep tor alpha gene polymorphisms by SSCP and its effect on rep roductive traits in Japanese flounder (Paralichthys olivaceus) [J].Comp Biochem Physiol B:Biochem Mol Biol,2008,150 (3):278-2831
[76]Rengmark A H, L ingaas F Genomic structure of the Nile tilap ia (O reochrom is niloticus) transferrin gene and a hap lotype associated with saltwater tolerance [J]. Aquac,2007,272 (1/4):146-1551
[77]Tatsuda K., Fujinaka K. Genetic mapping of the QTL affecting body weight in chickens using a F2 family. Br Poult Sci,2001,42(3):333-337.
[78]Wilhelm B. T., Marguerat S., Watt S., et al. Dynamic repertoire of a eukaryotic transcriptome surveyed at single-nucleotide resolution. Nature,2008,453(7199):1239-43.
[79]Elfstromc M, Gaffneyp M, Smithc T, et all Characterization of 12 single nucleotide polymorphisms in weathervane scallop[J]. Mol Ecol Notes,2005,5 (2):406-4091
[80]Aranishi, E., Okimoto, T. and Izumi, S. (2005 a) Identification of gadoid species (Pisces, Gadidae) by PCR-RFLP analysis. J. Appl. Genet.,46,69-73.
[81]Smith C T, Templin W D, Seeb J E, et al. Single nucleotide polymorphisms provide rapid and accurate estimates of the proportions of U.S. and Canadian Chinook salmon caught in Yukon River fisheries[J]. North American J Fish Manag,2005,25:944-953.
[82]Itoi S, Nakaya M, Kaneko G, et al. Rapid identification of eels Anguilla japonica and Anguilla anguilla by polymerase chain reaction with single nucleotide polymorphism-based specific probes[J]. Fish Sci,2005,71:1356-1364.
[83]Ye X., McLeod S., D. Elfick, Dekkers J. C., et al. Rapid identification of single nucleotide polymorphisms and estimation of allele frequencies using sequence traces from DNA pools. Poult Sci,2006,85:1165-1168.
[84]王爱民,石耀华.2003.中国马氏珠母贝遗传育种的现状与展望.农业生物技术学报,11(6)：547-553
[85]顾志峰,王嫣,石耀华等.2009.马氏珠母贝2个不同地理种群的形态性状和贝壳珍珠质颜色比较分析.渔业科学进展,30：79-86
[86]Beyer A., Bandyopadhyay S., Ideker T. Integrating physical and genetic maps:from genomes to interaction networks. Nat Rev Genet,2007,8(9):699-710.
[87]Ge H., Walhout A. J., Vidal M. Integrating 'omic' information:a bridge between genomics and systems biology. Trends Genet,2003,19(10):551-60.
[88]Germer S, Higuchi R (2003) Homogeneous allele-specific PCR in SNP genotyping. Methods in Molecular Biology,212,197-214.
[89]陈炜,张戈,张思仲.基于生物信息学的SNP候选位点搜寻方法[J].遗传,2001,23(2)：153-156
[90]Frazerk A, Ballingerd G, Coxd R, et al. A second generation human haplotype map of over3. 1 million SNPs[J]. Nature,2007,449:851-861
[91]Tenaillon M, Sawkinsmc, Longa D,et al. Patterns of DNA sequence polymorphism a 1 ong chromosome of maize (Zea mays ssp.maysL.)[J].ProcNatlAcad US A Sci,2001,9 8(16):9161-9166
[92]Sauvagec, Bieren, Lapegues, et al.Single nucleotide polymorphisms and their relationship to codonusage bias in the Pacific oyter Crassostreag igas [J].Gene,2007,406(1-2):13-22
[93]Lirh, Li Q, Konglf. Characterization of expressed sequence tag-derived single-nucleotide polymorphisms in the bay scallop Argopecten irradians [J]. Fish Sci 2009,75(6):1389-1400
[94]Zhang, L., Guo, X., Development and validation of single nucleotide polymorphism markers in the eastern oyster Crassostrea virginica Gmelin by mining ESTs and resequencing, Aquaculture (2010), doi:10.1016/j.aquaculture.2010.02.012
[95]吕敏芝2007.MS Excel在畜牧兽医统计中的应用(四)—卡方检验(X2检验).广东畜牧兽医科技,32(2)：30-32
[96]Richardi,Beckman J S. How neutral are synonymous codon mutations[J]. N at Genet, 1995,10:259.
[97]Carglll,M.,Altshuler,D.,Ireland,J.,Sklar,p.,Ardlie,K.Patil,N.,Shaw,N.,Lane,CR.,Lim.EP.,Kalya naraman,N.(1999)Charaeterization of single-nucleotide Polymorphisms in coding regions of human genes.NatGenet,22:231-38
[98]Nielsen,R.(2005)Molecular signatures of natural seleetion.Annu Rev Genet,39:197-218
[99]Francesca Capon,Richard C.Trembath,et al.(2004)A synonymous SNP of the corneodesmosin gene leads to increased mRNA stability and demonstrates association with psoriasis across diverse ethnic groups.Hum. Mol. Genet.13 (20):2361-2368.
[100]Giancarlo V. De Ferrari,et al.(2007)Common genetic variation within the Low-Density Lipoprotein Receptor-Related Protein 6 and late-onset Alzheimer's disease.PNAS May 29,vol.104 no.22:9434-9439
[101]Komar,A(2007) SNPs,Slient But Not Invisible,Science,315(5811),466-467(2007)