共查询到17条相似文献,搜索用时 78 毫秒
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以魁蚶为研究材料,建立了魁蚶的AFLP分子标记反应体系。对其分析过程中的酶切链接、预扩增、选择性扩增等试验条件进行了检测与筛选,得到了清晰的AFLP指纹图谱,为探讨魁蚶种群之间的遗传关系和遗传图谱的构建等方面提供了新的技术手段。 相似文献
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利用55对微卫星引物对珠江和海南长臀基因组DNA进行了扩增,有23对能扩增出清晰条带,其中有11对在珠江长臀中表现多态性,9对在海南长臀中表现多态。这些多态性位点的等位基因数在2~4之间,平均等位基因数2.91个。结果显示:珠江长臀的平均观测杂合度(Ho)是0.45,平均期望杂合度(He)是0.46,平均多态信息含量(PIC)为0.377;海南长臀的平均观测杂合度(Ho)和平均期望杂合度(He)分别是0.58、0.53,平均多态信息含量(PIC)为0.426。两群体间的遗传相似度为0.875、遗传距离为0.133。 相似文献
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大菱鲆AFLP分析体系的建立 总被引:5,自引:0,他引:5
本研究构建了大菱鲆扩增片段长度多态性(AFLP)分析体系,对DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤进行了分析。结果表明,用EcoRI/MseI双酶切、核心序列加3个选择性碱基的选择性扩增引物、1∶6(w∶w)的EcoRI端与MseI端选择性扩增引物比和20μl选择性扩增体积,可得到十分稳定的结果。所得产物经电泳和银染后可获得条带清晰、背景干扰小的图像。该体系的构建为AFLP技术在大菱鲆相关研究中的应用奠定了基础。 相似文献
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建立黑斑原鮡(Glyptosternum maculatum)AFLP反应体系,对DNA提取、双酶切、连接、预扩增、选择性扩增和银染效果进行了探索。结果表明,40μLTaqI酶切体系中65℃先酶切2h,EcoRI酶切和连接同时进行,EcoRI酶和T4DNA连接酶的用量均以2U为宜,温度与时间分别为37℃和3h,连接产物稀释10倍作为预扩增的模板。利用这种优化的反应体系,5对选择性扩增引物组合表现出较高的稳定性、清晰度和多态性,它们分别是:E-AAC/T-AAT、E-AAC/T-AAG、E-AAG/T-ACA、E-AAG/T-AAT和E-AAT/T-AGA。5对引物组合对96个样本扩增,共得到332条清晰可辨、可重复的条带,平均每对引物组合扩增出66.4条带,其中51条带是多态的。将AFLP技术用于黑斑原鮡的研究,通过优化体系的建立,为其种质资源的研究奠定基础。 相似文献
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本研究构建了西伯利亚鲟(Acipenser baeri)扩增片段长度多态性(AFLP)分析体系,分析了DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤。用7对引物对24尾西伯利亚鲟进行了AFLP分析,每对引物扩增的位点数在24~39之间,共扩增出了234个位点,多态性位点为116个,多态性比例为48.83%,平均Nei氏基因多样性指数H为0.4297,平均Shannon氏指数I为0.1233,个体间平均遗传距离为0.1872,表明西伯利亚鲟养殖群体的遗传多样性程度比较低,同时还构建了西伯利亚鲟的聚类图。 相似文献
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建立黑斑原鮡(Glyptosternum maculatum)AFLP反应体系,对DNA提取、双酶切、连接、预扩增、选择性扩增和银染效果进行了探索。结果表明,40μLTaqI酶切体系中65℃先酶切2h,EcoRI酶切和连接同时进行,EcoRI酶和T4DNA连接酶的用量均以2U为宜,温度与时间分别为37℃和3h,连接产物稀释10倍作为预扩增的模板。利用这种优化的反应体系,5对选择性扩增引物组合表现出较高的稳定性、清晰度和多态性,它们分别是:E-AAC/T-AAT、E-AAC/T-AAG、E-AAG/T-AC 相似文献
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Identification of growth‐related nucleotide polymorphism in cultured Pacific bluefin tuna,Thunnus orientalis 下载免费PDF全文
Yasuo Agawa Toshio Kaga Shigekazu Katayama Tokihiko Okada Yoshifumi Sawada 《Aquaculture Research》2017,48(7):3320-3328
Pacific bluefin tuna (PBF), Thunnus orientalis, is commercially one of the most important species of tuna. In this study, amplified fragment length polymorphism (AFLP) screening was conducted to find the growth‐related polymorphic DNA in cultured PBF. Fish hatched in 2007 were harvested at an age of 818–1994 days. They were categorized into superior, average and inferior growth groups, depending on their growth score at the time of harvest. On AFLP screening of 24 fish, with eight fish from each group, 215 polymorphic DNA fragments were observed. A second amplification, with EcoRI + ACC and MseI + CCC primers, generated a polymorphic fragment of 630 bp at a rate of 80.0% (n = 15) in the superior, 56.3% (n = 16) in the average and 20.0% (n = 15) in the inferior growth groups. Polymerase chain reaction (PCR) primers, which could amplify both AFLP‐positive and AFLP‐negative loci, were developed using the consensus sequence outside the AFLP target fragment. Eleven haplotypes were obtained by sequence analysis of the PCR product at the AFLP target loci. Among those, haplotype 1 was statistically significant in the superior and average growth groups and could be used as a molecular marker for distinguishing the individuals with superior and average growth from those with inferior growth. 相似文献
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长臀(Cranoglanis bouderius)和白甲鱼(Onychostoma sima)是目前北盘江主要放流鱼类,但这2种鱼的放流效果差异较大。由于鱼类的突进游速在一定程度上影响增殖放流的效果,故试验采用自制测试装置,用递增流速法测试了2种鱼的突进游速。结果显示,白甲鱼的突进游速大于长臀,且2种鱼的突进游速均随体长增加而近似线性增大;而相对突进游速则随着体长的增加而近似线性减小。研究结果可为北盘江鱼类增殖放流和放流效果评估,以及日后北盘江的拦鱼、诱鱼、集鱼船等鱼类资源保护措施的实施提供参考。 相似文献
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太空搭载卤虫遗传多样性及亲缘关系分析 总被引:1,自引:0,他引:1
为进一步探明空间诱变对卤虫休眠卵的影响效力,本研究选用4对AFLP选择性引物对太空诱变与地面对照组共59个卤虫个体进行了遗传多样性及亲缘关系分析。每对引物均获得清晰的扩增图谱,共检出222个清晰可辨的不同扩增片段,其中多态位点189个,占85.1%;UPGMA和NJ聚类分析图显示,来自空间诱变组的M5单独聚类,而其它58个个体聚为另一类,其他太空诱变组与地面对照组的个体交叉聚类,地面对照和太空诱变组遗传差异不明显。研究结果表明:实验用卤虫遗传多样性相当丰富为优良品系的选育提供了很好的基础,但两组间及组内的遗传差异主要是由个体间本身差异而导致的。 相似文献
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利用显微介导远缘杂交技术将河蟹总DNA直接导入镜鲤受精卵内,再利用AFLP分子标记技术检测显微介导河蟹基因镜鲤的外源DNA。在30对AFLP引物中,有3对引物扩增出供体基因片段,即在显微介导河蟹基因镜鲤中均有和河蟹基因相同而对照镜鲤没有的条带。对6尾阳性显微介导河蟹基因镜鲤的扩增产物进行回收、克隆和测序验证。结果表明:阳性显微介导河蟹基因在镜鲤中均含有供体基因目的片段。本研究在分子和基因水平上验证了河蟹总DNA可通过显微介导方式整合到镜鲤基因组中,为显微介导外源总DNA转化技术的应用提供新的思路。 相似文献
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The genetic variability among Streptococcus agalactiae isolates recovered from fish was characterized using single-stranded conformation polymorphism (SSCP) analysis of the intergenic spacer region (ISR), and amplified fragment length polymorphism (AFLP) fingerprinting. A total of 46 S. agalactiae cultures isolated from different fish species and geographic origins as well as related reference strains were included in the study. ISR-SSCP divided the S. agalactiae isolates analysed into five distinct genotypes. Genotype 1 grouped all Kuwait isolates while genotype 4 clustered the majority of non-Kuwait isolates (USA, Brazil and Honduras). AFLP analysis offered a higher resolution level by dividing the isolates into 13 different genotypes. Two different AFLP profiles were identified within the Kuwait isolates. When data from both ISR-SSCP and AFLP were combined through a multidimensional analysis (MDS), a good correlation between geographical origin and genotypes was observed. Both AFLP and ISR-SSCP revealed genetic differences between S. agalactiae isolates from fish. While AFLP offered a higher resolution, ISR-SSCP also provided valid information being a simpler and faster method. 相似文献