Simmen F A, Dolecki G J, Carlos R, Mandel M, Humphreys T
DNA. 1985 Oct;4(5):385-93. doi: 10.1089/dna.1985.4.385.
Ribosomal gene sequences (rDNA) were isolated from the genomic DNA of the Hawaiian sea urchin Tripneustes gratilla by cloning in plasmid and phage vectors. The rRNA coding regions in four clones were localized by hybridizing Southern transfers of endonuclease-digested DNA with 32P-labeled 26S rRNA and 32P-labeled 18S rRNA. Three of the rDNA clones were isolated from a library of DNA from a single sea urchin and represent the two major types of rDNA repeats present in that individual's genome. Both repeat types appear identical within their rRNA coding regions but are dissimilar in an area of the nontranscribed spacer (NTS) adjacent to the 3' end of the 26S coding region. The cloned rDNA repeats were shorter than their genomic counterparts due to deletions occurring within internally repetitious NTS domains, probably as a result of unequal recombination during phage propagation.
通过在质粒和噬菌体载体中克隆,从夏威夷海胆波纹唇鱼的基因组DNA中分离出核糖体基因序列(rDNA)。通过用32P标记的26S rRNA和32P标记的18S rRNA与核酸内切酶消化的DNA的Southern转移杂交,确定了四个克隆中的rRNA编码区域。三个rDNA克隆是从单个海胆的DNA文库中分离出来的,代表了该个体基因组中存在的两种主要类型的rDNA重复序列。两种重复类型在其rRNA编码区域内看起来相同,但在与26S编码区域3'端相邻的非转录间隔区(NTS)区域中不同。由于在内部重复的NTS结构域内发生缺失,克隆的rDNA重复序列比其基因组对应物短,这可能是噬菌体繁殖期间不等重组的结果。