Zhuo L, Sajdak S L, Phillips R B
Department of Biological Sciences, University of Wisconsin-Milwaukee 53201.
Genome. 1994 Aug;37(4):664-71. doi: 10.1139/g94-094.
Intraspecific variation in the sequence of the transcribed spacer regions of the ribosomal DNA (rDNA) in lake trout was examined by restriction mapping and sequencing of these regions amplified by the polymerase chain reaction. The length of the first internal transcribed spacer region (ITS-1) was 566 bases and the second internal transcribed spacer region (ITS-2) was 368 bases in lake trout. When the 1.4-kb region including the ITS-1, the 5.8S coding region, and the ITS-2 was amplified from 12 individuals from four populations and digested with eight different enzymes only one intraindividual polymorphism was found that occurred in each population. When the amplified ITS-1 region was sequenced from an additional 10 individuals from five populations, no interindividual variation was found in the sequence. A 6-kb portion of the rDNA repeat unit including 1.6 kb of the 18S coding region, the 5' external spacer region (5' ETS), and part of the adjacent intergenic spacer was cloned and a restriction map was prepared for these regions in lake trout. No intraspecific variation was found in the region adjacent to the 18S rDNA, which includes the 5' ETS, although intraspecific and intraindividual length variation was found in the intergenic spacer region 3-6 kb from the 18S. Sequencing of a 609-b segment of the 5' ETS adjacent to the 18S coding region revealed the presence of two 41-b repeats. The 198-b sequence between the repeats had some similarity to the 18S coding region of other fishes. Primers were designed for amplification of 559 b of the 5' ETS using the polymerase chain reaction.(ABSTRACT TRUNCATED AT 250 WORDS)
通过对核糖体DNA(rDNA)转录间隔区序列进行限制酶切图谱分析和聚合酶链反应扩增区域测序,研究了湖红点鲑rDNA转录间隔区序列的种内变异。湖红点鲑的第一个内部转录间隔区(ITS-1)长度为566个碱基,第二个内部转录间隔区(ITS-2)长度为368个碱基。当从四个种群的12个个体中扩增出包含ITS-1、5.8S编码区和ITS-2的1.4 kb区域并用八种不同的酶进行消化时,仅发现每个种群中都出现了一种个体内多态性。当从五个种群的另外10个个体中对扩增的ITS-1区域进行测序时,在序列中未发现个体间变异。克隆了rDNA重复单元的6 kb部分,包括1.6 kb的18S编码区、5'外部间隔区(5' ETS)和相邻基因间隔区的一部分,并为湖红点鲑的这些区域制备了限制酶切图谱。在与18S rDNA相邻的区域(包括5' ETS)中未发现种内变异,尽管在距18S 3 - 6 kb的基因间隔区发现了种内和个体内长度变异。对与18S编码区相邻的5' ETS的609 bp片段进行测序,发现存在两个41 bp的重复序列。重复序列之间的198 bp序列与其他鱼类的18S编码区有一些相似性。设计了引物,用于通过聚合酶链反应扩增5' ETS的559 bp。(摘要截短至250字)