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Mining expressed sequences for single nucleotide polymorphisms in Pacific abalone Haliotis discus hannai
Qi, Haigang1,2; Liu, Xiao1; Zhang, Guofan1; Wu, Fucun1; Liu, X, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China
2009-09-30
发表期刊AQUACULTURE RESEARCH
ISSN1355-557X
卷号40期号:14页码:1661-1667
文章类型Article
摘要Although single nucleotide polymorphisms (SNPs) are important resources for population genetics, pedigree analysis and genomic mapping, such loci have not been reported in Pacific abalone so far. In this study, a bioinformatics strategy was adopted to discover SNPs within the expressed sequences (ESTs) of Pacific abalone, Haliotis discus hannai, and furthermore, polymerase chain reaction direct sequencing (PCR-DS) and allele-specific PCR (AS-PCR) were used for SNPs detection and genotype scoring respectively. A total of 5893 ESTs were assembled and 302 putative SNPs were identified. The average density of SNPs in ESTs was 1%. Fifty-two sets of sequencing primers were designed from SNPs flanking ESTs to amplify the genomic DNA, and 13 could generate products of expected size. Polymerase chain reaction direct sequencing of the amplification products from pooled DNA samples revealed 40 polymorphic SNP loci. Using a modified tetra-primer AS-PCR, seven mitochondrial and six nuclear SNPs were typed and characterized among 37 wild abalones. In conclusion, it is feasible to discover SNPs from number limited ESTs and the AS-PCR as a simple, robust and reliable assay could be a primary method for small- and medium-scale SNPs detection in abalones as well as other non-model organisms.; Although single nucleotide polymorphisms (SNPs) are important resources for population genetics, pedigree analysis and genomic mapping, such loci have not been reported in Pacific abalone so far. In this study, a bioinformatics strategy was adopted to discover SNPs within the expressed sequences (ESTs) of Pacific abalone, Haliotis discus hannai, and furthermore, polymerase chain reaction direct sequencing (PCR-DS) and allele-specific PCR (AS-PCR) were used for SNPs detection and genotype scoring respectively. A total of 5893 ESTs were assembled and 302 putative SNPs were identified. The average density of SNPs in ESTs was 1%. Fifty-two sets of sequencing primers were designed from SNPs flanking ESTs to amplify the genomic DNA, and 13 could generate products of expected size. Polymerase chain reaction direct sequencing of the amplification products from pooled DNA samples revealed 40 polymorphic SNP loci. Using a modified tetra-primer AS-PCR, seven mitochondrial and six nuclear SNPs were typed and characterized among 37 wild abalones. In conclusion, it is feasible to discover SNPs from number limited ESTs and the AS-PCR as a simple, robust and reliable assay could be a primary method for small- and medium-scale SNPs detection in abalones as well as other non-model organisms.
关键词Snps Est Pacific Abalone Haliotis Discus Hannai Pcr-ds As-pcr
学科领域Fisheries
DOI10.1111/j.1365-2109.2009.02269.x
URL查看原文
收录类别SCI
语种英语
WOS记录号WOS:000270124500011
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被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/1864
专题海洋生物技术研发中心
通讯作者Liu, X, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China
作者单位1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
第一作者单位中国科学院海洋研究所
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Qi, Haigang,Liu, Xiao,Zhang, Guofan,et al. Mining expressed sequences for single nucleotide polymorphisms in Pacific abalone Haliotis discus hannai[J]. AQUACULTURE RESEARCH,2009,40(14):1661-1667.
APA Qi, Haigang,Liu, Xiao,Zhang, Guofan,Wu, Fucun,&Liu, X, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China.(2009).Mining expressed sequences for single nucleotide polymorphisms in Pacific abalone Haliotis discus hannai.AQUACULTURE RESEARCH,40(14),1661-1667.
MLA Qi, Haigang,et al."Mining expressed sequences for single nucleotide polymorphisms in Pacific abalone Haliotis discus hannai".AQUACULTURE RESEARCH 40.14(2009):1661-1667.
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