摘要
分析了缢蛏(Sinonovacula constricta)基因组文库中微卫星序列的特性及微卫星引物的遗传特征。采用磁珠富集珐和PCR筛选法,以CA15为探针,快速分离含有微卫星序列的阳性克隆。结果表明,在筛选的210个白色菌落中,得到58个微卫星序列,阳性克隆率为27.6%,核心重复序列为两碱基和三碱基的微卫星居多,其中完美型占39.7%,非完美型39.7%,复合型占10.3%,非完美型与复合型共存的占10.3%。微卫星重复次数主要集中在20～40次之间,占66%,最高为63次。除探针中使用的CA重复单元外,还观察到AG/TC,AT/TA,AAC/TTG,CAA/GTT,TGA,AAT,AGAA,GTTT/CAAA,AAAT/TTTA,TTTC,CCTA,TCTA,TCCC,TGCC,GTTTCT,CTGTT,CACACC,TATACA,TTTTG的重复序列。利用Primer Premier5.0设计引物24对,经过初步筛选得到适合的微卫星引物18对,为今后未知微卫星标记的开发,群体遗传结构的分析,优良品种的选育提供基础。
The microsatellite sequences and microsatellite primers were analyzed in razor clam（ Sinonovacula constricta ）. The positive clones of microsatellites were isolated using the magnetic bead hybridization selection method and PCR selection method with the（CA）is prone. A total of 210 white colonies were screened using a PCR-based technique, and there were 58 microsatellite repeat motifs by sequencing（27.6% of white colonies screened）. Besides the motif of CA contained in the oligoprobe, we also found other 19 types of microsatellite repeats including AG/TC, AT/TA, AAC/TTG, CAA/GTT, TGA, AAT, AGAA, GTTT/CAAA, AAAT/ TTTA, TTTC, CCTA, TCTA, TCCC, TGCC, GTTTCT, CTGTT, CACACC, TATACA and TTTTG. According to Weber（1990）, the microsatellite sequences obtained could be categorized Structurally into perfect repeats（39.7 % ）, imperfect repeats（39.7 % ）, and compound repeats（20.6 % ）. Among the microsatellite repeats, 20--40 repeats arrays were most abundant, accounting for 66%. The largest length of microsatellites was 63 repeats. Finally, 24 microsatellite primers were designed by software Premier Primer 5.0 and 18 pairs were applied for the population genetic studies. They may be applied for the population genetic diversity and developing genetic linkage maps.
出处
《中国海洋大学学报（自然科学版）》
CAS
CSCD
北大核心
2008年第5期733-738,共6页
Periodical of Ocean University of China
基金
国家高技术研究发展计划项目(2006AA10A410)
上海市重点学科建设项目(Y1101)资助
关键词
缢蛏
微卫星
磁珠富集
PCR
Sinonovacula constricta
microsatellite
enrichment by magnetic beads
PCR
