利用微卫星 DNA 分析鉴定虹鳟细胞系。
Characterization of rainbow trout cell lines using microsatellite DNA profiling.
机构信息
Department of Zoology, University of Guelph, Guelph, N1G 2W1, Canada.
出版信息
Cytotechnology. 2001 Nov;37(3):143-51. doi: 10.1023/A:1020516804173.
Ten microsatellite loci (Omy27DU,Omy325(A3)UoG, OmyFGT5TUF,OmyFGT14TUF, OmyFGT15TUF,OmyFGT23TUF, Omy77DU,Ssa20.19NUIG, Ots1BML, andOne18ASC) were amplified using the polymerase chain reaction to create genetic profiles for nine cell lines (RTG-2, RTH-149,RTL-W1,RTgill-W1, RTS-11, RTS-34st, RTP-2, RTP-91E and RTP-91F) from rainbow trout(Oncorhynchus mykiss) and one cell line (CHSE-214) from Chinook salmon (O. tschawytscha). A cell line (PHL) from anon-salmonid, the Pacific herring (Clupea harengus pallasi), was included as a control. The ten loci clearly revealed the uniqueness of each cell line, except for two cell lines (RTP-91E andRTP-91F) from the same fish. RTP-91E and RTP-91F were identical at all loci except Ssa20.19NUIG. The most useful locus for demonstrating uniqueness was Ots1BML. The information was used to demonstrate that an uncharacterized rainbow trout cell line (Clone 1A)was in fact CHSE-214, illustrating the usefulness of multiplexed microsatellites for the creation of genetic profiles for salmonid cell lines and for the testing of cell line cross-contamination.
使用聚合酶链反应扩增了 10 个微卫星位点（Omy27DU、Omy325(A3)UoG、OmyFGT5TUF、OmyFGT14TUF、OmyFGT15TUF、OmyFGT23TUF、Omy77DU、Ssa20.19NUIG、Ots1BML 和 One18ASC），为来自虹鳟鱼（Oncorhynchus mykiss）的 9 个细胞系（RTG-2、RTH-149、RTL-W1、RTgill-W1、RTS-11、RTS-34st、RTP-2、RTP-91E 和 RTP-91F）和一条来自奇努克鲑鱼（O. tschawytscha）的细胞系（CHSE-214）创建遗传图谱。一个非鲑鱼的细胞系（PHL），太平洋鲱鱼（Clupea harengus pallasi），被用作对照。除了来自同一条鱼的两个细胞系（RTP-91E 和 RTP-91F）外，这 10 个位点清楚地显示了每个细胞系的独特性。RTP-91E 和 RTP-91F 在所有位点上除了 Ssa20.19NUIG 外都是相同的。最有用的位点是 Ots1BML，用于证明独特性。该信息用于证明一个未被描述的虹鳟鱼细胞系（Clone 1A）实际上是 CHSE-214，这说明了复用微卫星对鲑鱼细胞系遗传图谱的创建和细胞系交叉污染测试的有用性。
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