Gale K, Crampton J
Wolfson Unit of Molecular Genetics, Liverpool School of Tropical Medicine, UK.
Eur J Biochem. 1989 Nov 6;185(2):311-7. doi: 10.1111/j.1432-1033.1989.tb15117.x.
The characterisation of the ribosomal genes of the mosquito, Aedes aegypti, is described. Preliminary experiments using a cloned Drosophila ribosomal DNA (rDNA) repeat to probe Southern transfers of Ae. aegypti genomic DNA has indicated that the rDNA repeat of Ae. aegypti is 9.0 kb in length and that individual rDNA repeats exhibit a high degree of homogeneity with respect to length and the position of restriction enzyme recognition sites within the rDNA. The preliminary mapping data together with partial digestion experiments demonstrate that, as in all other higher eukaryotes, the rDNA repeats are arranged in a head-to-tail, tandemly repeating manner. The restriction mapping of cloned rDNA repeats confirmed the largely uniform length of the Ae. aegypti rDNA repeat and provided a more detailed physical map of the DNA. A restriction site polymorphism was detected in one clone (Aar9) which contains an extra HincII site, which is not present in three other clones studied (Aar1, Aar3, or Aar7). Transcription mapping has allowed the allocation of identities to the various restriction fragments and the approximate positioning of the transcription unit. The estimate of rDNA repeat copy number in Ae. aegypti (approximately 500 copies per haploid genome) is similar to the estimate reported for the closely related species, Aedes albopictus, of 430 copies per haploid genome. Ribosomal DNA thus comprises approximately 0.6% of the total Ae. aegypti genome. Analysis of the variation of the rDNA repeat unit both within individual mosquitoes and between strains of Ae. aegypti, has severed to confirm the remarkable homogeneity of the rDNA repeat unit in this insect.
本文描述了埃及伊蚊核糖体基因的特征。使用克隆的果蝇核糖体DNA(rDNA)重复序列对埃及伊蚊基因组DNA的Southern转移进行探测的初步实验表明,埃及伊蚊的rDNA重复序列长度为9.0 kb,并且各个rDNA重复序列在长度以及rDNA内限制酶识别位点的位置方面表现出高度的同质性。初步的图谱数据以及部分消化实验表明,与所有其他高等真核生物一样,rDNA重复序列以头对尾的串联重复方式排列。克隆的rDNA重复序列的限制酶图谱证实了埃及伊蚊rDNA重复序列的长度基本一致,并提供了更详细的DNA物理图谱。在一个克隆(Aar9)中检测到一个限制酶切位点多态性,该克隆含有一个额外的HincII位点,而在其他三个研究的克隆(Aar1、Aar3或Aar7)中不存在此位点。转录图谱分析已确定了各个限制片段的身份以及转录单元的大致位置。对埃及伊蚊rDNA重复序列拷贝数的估计(每个单倍体基因组约500个拷贝)与报道的近缘物种白纹伊蚊的估计值相似,即每个单倍体基因组430个拷贝。因此,核糖体DNA约占埃及伊蚊基因组总量的0.6%。对单个蚊子体内以及埃及伊蚊不同品系之间rDNA重复单元变异的分析,进一步证实了该昆虫rDNA重复单元的显著同质性。