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| 1 | 诸氏鲻虾虎鱼转录组序列中微卫星标记的初步筛选及特征分析显示文摘旨在为大规模开发诸氏鲻虾虎鱼微卫星标记,采用高通量测序技术,对诸氏鲻虾虎鱼肝脏转录组进行了测序。结果共获得47 979条Unigenes,利用微卫星查找程序在47 979条Unigenes中共获得6 225个微卫星位点(12.97%),平均每7.02 kb就出现1个微卫星位点。6 225个微卫星位点由226种重复基序组成,主要分布在三、四和五碱基重复类型中。在数量上,单碱基重复类型微卫星位点最多,占42.49%,二碱基和三碱基重复类型所占比例相似,分别为25.22%和26.27%,四、五、六重复类型较少,合计占6.03%。单碱基重复序列中最多的类型为A/T,二碱基重复序列中以AG/CT重复单元为主,三碱基重复序列中以AGC/TCG为优势类型。挑选部分二、三和四单元重复类型微卫星序列,共设计76对引物,可稳定扩增出目的条带的有55对,其中32对具有多态性。结果表明,利用诸氏鲻虾虎鱼转录组数据可快速大量开发微卫星标记。 | 蔡磊 余露军 陈小曲 叶惠欣 陈琳 李建军 | 2015 | 生物技术通报2015，31，9： | 16 |
| 2 | 印尼虎鱼RAD-seq数据中微卫星标记的初步筛选及特征分析显示文摘采用RAD-seq(Restriction-site associated DNA sequencing)对印尼虎鱼(Datnioides pulcher)进行简化基因组测序,获得13308 806个高质量干净读长(HQ clean reads),利用SSR搜索软件在所得序列中共检测到26359个SSR位点,由496种重复基序组成,主要分布在三、四、五碱基重复类型中,单碱基重复序列中最多的类型为A/T,二碱基重复序列中以AC/GT和AG/GT重复单元为主,三碱基重复序列中以AGG/CCT、AGC/CTG、AGG/CCT为优势类型;在数量上,二碱基重复类型SSR位点最多(19492个),占总SSR位点的73.95%;除了单核苷酸类型外,SSR基序的重复次数主要集中在4-9之间,多态性SSR数目为2 783个。根据SSR位点两翼序列,利用Primer5.0成功设计出20 066对SSR引物,引物设计成功率为76.13%,依据测序数据中的重复类型和重复数,随机选择100个二至五碱基重复类型SSR在6个虎鱼样本中验证其可用性和多态性,有73对引物通过扩增可获得有效目的片段,其中20个SSR验证为具有多态性。开发所得SSR序列可用于印尼虎鱼及其近缘物种遗传分析、遗传图谱构建以及分子标记辅助育种等工作,同时也证实了利用简化基因组测序技术开发SSR标记和发现样品间具有多态性的SSR标记的可能性。 | 屈政委 宋红梅 汪学杰 刘奕 牟希东 刘超 胡隐昌 | 2019 | 淡水渔业2019，49，4： | 7 |
| 3 | 基于胡鲇科和鲇科ESTs序列初步筛选兰州鲇EST-SSR分子标记显示文摘利用SSRHunter软件对现已公布的鲇形目(Siluriformes)中胡鲇科(Clariidae)和鲇科(Siluridea)共2002条EST(Expressed Sequence Tag)序列筛选兰州鲇(Silurus lanzhouensis)微卫星标记,并分析其序列丰度和分布特征。结果共发现209个微卫星序列,占整个ESTs数据库的10.44%;其中三碱基重复序列数量最多,占ESTs数据库中发现微卫星序列总数的35.62%;二、四、五、六碱基重复序列分别占微卫星序列总数的28.31%、25.57%、7.76%、2.74%。根据侧翼序列,设计125对引物,随机选取58对进行扩增鉴定及多态性筛选。结果显示29对引物能稳定扩增,16对引物具有多态性,每个位点等位基因数2-7个(平均3.75),观测杂合度(Ho)0.133 2-0.732 2(平均0.405 1),期望杂合度(He)0.198 2-0.848 3(平均0.560 2),多态信息含量(PIC)0.124 1-0.794 5(平均0.506 5)。研究表明,借用已有鲇形目鱼类ESTs数据库筛选到的兰州鲇微卫星标记质量较好,可为兰州鲇种质资源保护、微卫星连锁图谱构建、经济性状的QTL定位及分子标记辅助育种奠定基础。 | 魏大为 连总强 吴旭东 王发新 李力 俞兆曦 杨忠礼 | 2015 | 淡水渔业2015，45，5： | 3 |
| 4 | Preliminary Screening and Characterization of Microsatellite Markers in RAD-seq Data of Datnioides pulcher显示文摘[Objective]The paper was to develop the microsatellite markers of Datnioides pulcher,and to screen polymorphic SSR primers.[Method]D.pulcher was performed reduced-representation genome sequencing using RAD-seq(Restriction-site associated DNA sequencing).[Result]A total of 13308806 high-quality clean reads(HQ clean reads)lengths were obtained and 26359 simple sequence repeats(SSR)loci were detected in the obtained sequence by SSR search software,which consisted of 496 repeat motifs.The most common types of repeat were A/T in single base repeat sequence,and AC/GT and A G/GT repeat units in di-base repeat sequence.AGG/CCT,AGC/CTG and AGG/CCT were the dominant types in tri-base repeat sequence.In quantity,the di-base repeat type SSR loci were the largest(19492),accounting for 73.95%of the total SSR loci.Except for mono-nucleotide type,the repetition frequency of SSR loci was mainly ranged from 4 to 9,and the number of polymorphic SSR loci was 2783.About 20066 pairs of SSR primers were successfully designed with Primer 5.0.The success rate of primer design was 76.13%.According to the repetition type and repetition number in the sequencing data,100 di-to penta-base repetition type SSRs were randomly selected and validated by PCR and polyacrylamide gel in six D.pulcher samples.A total of 73 pairs of primers were amplified by electrophoresis,and 20 of them were polymorphic.[Conclusion]The developed SSR sequences can be used for genetic analysis,genetic map construction and molecular marker assisted breeding of D.pulcher and its related species. | Qu Zhengwei Song Hongmei Wang Xuejie Liu Yi Mu Xidong Liu Chao Hu Yinchang | 2020 | Animal Husbandry and Feed Science2020，12，2： | 0 |
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