| 摘要: |
| 应用6个微卫星标记在奉化泥蚶种群上进行扩增,以检测其遗传信息。扩增结果表明,引物所扩增出的等位基因数目变化从3个到7个,平均等位基因数目为5.4。引物的多态性信息含量范围为0.111-0.814。最大的观察杂合度Ho为0.690,最小为0.121。期望杂合度He范围为0.117-0.848。位点TMP22显著偏离了哈代-温伯格平衡。6个引物在18个马来西亚泥蚶上进行扩增,结果只有TMP18和TMP36在马来西亚模板上有扩增产物。 |
| 关键词: 泥蚶 微卫星 等位基因 |
| DOI：10.11693/hyhz200806018 |
| 分类号: |
| 基金项目:国家高技术研究发展计划(863计划)项目资助,2006AA10A410号;国家科技支撑计划项目资助,2007BAD43B09号;浙江省重大科技攻关计划项目,2006C12013号 |
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| ISOLATION AND CHARACTERISTICS OF SIX MICROSATELLITE PRIMERS OF TEGILLARCA GRANOSA |
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GU Xiao-Ying,ZENG Qing-Guo,YOU Zhong-Jie,LIN Zhi-Hua
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1.Key Laboratory of Applied Marine Biotechnology (Ningbo University), Ministry of Education, Ningbo, 315211;2.The Ocean and Fishery Institute of Ningbo, Ningbo, 315012;3.Zhejiang Mariculture Research Institute, Wenzhou, 325005
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| Abstract: |
| Six microsatellite loci were isolated and then used to investigate the genetic diversity of blood clam (Tegillarca granosa) collected form Fenghua, Zhejiang Province. The microsatellite primers were first screened in a polymerase chain reaction through a temperature gradient to determine best annealing temperature. The functional primers were then selected for population amplification. PCR was performed in a 25μl reaction system as follows: 1×PCR buffer, 2.0mm MgCl2, 0.2mm dNTPs, l0pmol per primer, 0.65U Taq polymerase, and 10ng genomic DNA. Reaction took place in a PCR amplifier (Mastercycler Eppendorf) in program of: 5min at 94°C for denaturation, 30 cycles of 94°C 1 min, 30 s at annealing temperature, 72°C extension for 45s, and a final extension for 7min at 72°C. The PCR products were afterwards electrophoresed on 8% polyacrylamide gel and detected in silver staining method. Polymorphic information content (PIC) was calculated with CERVUS 2.0 software. The expected heterozygosity (He) and observed heterozygosity Ho) was estimated with GENEPOP 4.0.7. The allele number per loci varied from 3 to 7, PIC value was 0.111 to 0.814. The mean Ho and He varied from 0.121 to 0.690 and 0.117 to 0.848, respectively. Primer amplification showed that some microsatellite loci isolated from the samples of continental China were absent in those of Malaysia. |
| Key words: Blood clam Tegillarca granosa, Microsatellite, Allele |
