Identification of Clones Induced by Chromosome Manipulation in Ayu Plecoglossus altivelis by DNA Fingerprinting with RI and Non-RI Labelled Probes
Identification of Clones Induced by Chromosome Manipulation in Ayu Plecoglossus altivelis by DNA Fingerprinting with RI and Non-RI Labelled Probes
复制标题
通过 RI 和非 RI 标记探针进行 DNA 指纹图谱鉴定香鱼染色体操作诱导的克隆
DOI：
10.2331/fishsci.61.909
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发表时间：
1995
影响因子：
1.9
通讯作者：
A. Tsujimura
中科院分区：
文献类型：
--
作者：
M. Takagi;N. Taniguchi;Motoshi Yamasaki;A. Tsujimura
应用DNA指纹图谱(DNA-FP)对高山羊舌兰(Plecoglossus Altivelis)的第一次卵裂抑制所诱导的克隆进行了确认和鉴定。所用样本为2个纯合子无性系、3个连续纯合无性系和2个杂合子无性系。我们检测了合适的酶/探针组合，并比较了放射性同位素(RI)标记的探针和非放射性同位素(Non-RI)标记的探针，以获得清晰的DNA-FP。YNZ 22、33.15和33.6探针分别与Hae III、Msp I和Hinf I限制性内切酶反应，获得清晰的DNA指纹图谱。在这些DNA-FP中，高分子量区和低分子量区的条带在整个膜上明显分开。用非RI标记的33.15探针获得的DNA-FP条带与RI方法得到的DNA-FP条带一样强烈和清晰。所有的片段在同一克隆系中的个体之间是共享的，但克隆系之间检测到的条带模式明显不同。证实了DNA-FP技术在无性系鉴定中的有效性。非RI标记探针的DNA-FP图谱适合在近亲繁殖和个体及家系鉴定研究中广泛应用。
DNA fingerprint (DNA-FP) was applied for confirmation and identification of clones induced by the suppression of the first cleavage in ayu, Plecoglossus altivelis. The samples used were 2 homozygous clonal lines, 3 successive homozygous clonal lines and 2 heterozygous clonal lines . We examined suita ble enzyme/probe combinations and compared radioisotope (RI) and non-radioisotope (Non-RI) labelled probes to get clear DNA-FP. The clear DNA fingerprints were obtained by using YNZ 22 , 33.15 and 33.6 probes with Hae III, Msp I and Hinf I restriction enzymes. In these DNA-FPs, the bands were separated distinctly throughout the membrane in both higher and lower molecular weight regions. The DNA-FP bands obtained with the Non-RI labelled 33.15 probes were as intense and clearly separated as those obtained with the RI method. All the fragments were shared among individuals within the same clonal line , but the pattern of bands detected was significantly different between clonal lines. It was confirmed that DNA-FP is effective for the identification of clonal lines. DNA-FP pattern by Non-RI labelled probe would be suitable for wide applications in studies on inbreeding and identifica tion of individuals and families.