中国对虾微卫星标记的筛选

IOCAS-IR > 海洋环流与波动重点实验室

	中国对虾微卫星标记的筛选
其他题名	Isolation of Microsatellite Markers from Chinese Shrimp
	徐鹏
学位类型	博士
	2002
学位授予单位	中国科学院海洋研究所
学位授予地点	中国科学院海洋研究所
学位专业	海洋生物学
关键词	中国对虾微卫星微卫星标记
摘要	该文运用PCR法在中国明对虾（Fenneropenaeuschinensis）的小片段基因组文库中快速筛选含有微卫星序列的重组阳性克隆.利用生物信息学的方法,使用Repeatmasker软件对中国对虾ESTs数据库中10446条EST序列进行了微卫星序列的筛选.在发现的微卫星序列中选取了19条微卫星序列设计了引物,在16对能够扩增出产物的引物中,经过PCR扩增、聚丙烯酰胺凝胶电泳和硝酸银染色,最终共筛选出有多态性PCR产物的微卫星引物11对,其中适合进行微卫星分析的有8对.在中国对虾20个左右样本的8个微卫星位点上,等位基因的数目从5到15不等,等位基因的大小分布在165bp到305bp之间,基本符合引物设计时的理论产物长度.
其他摘要	Recombinant positive clones containing microsatellite sequences of Chinese shnmp(Fenneropeneaus chinensis) were obtained through rapid screening small-size fractionated genomic libraries with PCR technique. The primers used in PCR were T7/SP6 promotor primers located at both sides of multiple clone site of pGEM-3zf (+) and STR primers: STR1: 5'-ATATATATATATAT-3' and STR2: 5'-TCTCTCTCTCTCTC-3' designed according to core repeats motifs of microsatellites. Thirty-one microsatellites of Chinese shrimp F. chinensis were obtained after sequencing for the first time. Among the 31 microsatellites in 18 positive recombinant positive clones are 23 perfect ones(74%), 2 imperfect(7%), 1 compound perfect (3%), 5 compound imperfect (16%). By using the on-line bioinformatics software Repeatmasker on internet, the ESTs database of Chinese shrimp was screened. 229 microsatellite sequences were obtained from 10446 ESTs sequences, about 2.19% in whole database. A total 146 dinucleotide repeats and 58 trinucleotide repeats were found in these 229 microsatellite sequences and most were perfect repeats, with (AT)n in dinucleotide repeats and (TAA),, in trinucleotide repeats being the most common, about 28.82% and 17.03% in the total microsatellite sequences respectively. Nineteen primer pairs were designed from microsatellites using computer program Primer Premier (Version 5.00). Of the 16 primers that produced product, Eleven primers were polymorphic in about 20 Chinese shrimp samples and Eight primers were suitable for microsatellite analysis. In these eight microsatellite loci, the alleles numbers distributed from 5 to 15, and the range of these alleles were 165 to 305bp and similar with their predicted size. The expected heterozygosity and the PIC values of them distributed from 0.59 to 0.89 and 0.56 to 0.88, respectively. These values showed that these 8 microsatellite loci were suitable for genetic analysis.
页数	74
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/1177
专题	海洋环流与波动重点实验室
推荐引用方式 GB/T 7714	徐鹏. 中国对虾微卫星标记的筛选[D]. 中国科学院海洋研究所. 中国科学院海洋研究所,2002.

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