从人类丝虫寄生虫马来布鲁线虫基因组中分离并鉴定一种重复DNA元件。
Isolation and characterization of a repetitive DNA element from the genome of the human filarial parasite, Brugia malayi.
作者信息
Natarajan S, Werner C, Cameron M, Rajan T V
机构信息
Department of Pathology, University of Connecticut Health Center, Farmington.
出版信息
Mol Biochem Parasitol. 1990 Nov;43(1):39-49. doi: 10.1016/0166-6851(90)90128-9.
The genome of the human filarial parasite Brugia malayi contains at least two major repetitive DNA elements. One, referred to as the HhaI family, consists of 10(4)-10(5) tandemly arrayed copies per haploid genome of a monomer of 322 base pairs and does not contain a cleavage site for the restriction endonuclease MboI. We constructed a library of MboI-digested genomic B. malayi DNA in BamHI-cut M13mp18 resulting in the exclusion of the HhaI repeat family from the library. Hybridization of this genomic library with nick-translated genomic DNA yielded several copies of a repeat family which we have named the BmMboI family. From sequence analysis of more than 50 monomers, which differ from each other in sequence and length, we have been able to divide the monomers into several regions based on the level of sequence conservation. Southern blot analyses of B. malayi genomic DNA digested with a variety of restriction endonucleases and probed with the isolated repeat demonstrate multiple bands of varying sizes except with HindIII-cut DNA, where the repeat is found only in very high-molecular-weight DNA.
人类丝虫寄生虫马来布鲁线虫的基因组包含至少两种主要的重复DNA元件。一种被称为HhaI家族，每个单倍体基因组中有10⁴ - 10⁵个322个碱基对的单体串联排列拷贝，并且不包含限制性内切酶MboI的切割位点。我们构建了一个用MboI消化的马来布鲁线虫基因组DNA文库，该文库位于用BamHI切割的M13mp18中，从而使文库中排除了HhaI重复家族。用缺口平移的基因组DNA对该基因组文库进行杂交，得到了一个重复家族的几个拷贝，我们将其命名为BmMboI家族。通过对50多个彼此序列和长度不同的单体进行序列分析，我们能够根据序列保守程度将单体分为几个区域。用多种限制性内切酶消化马来布鲁线虫基因组DNA并用分离出的重复序列进行探针杂交分析，结果显示除了用HindIII切割的DNA外，有大小各异的多条带，在用HindIII切割的DNA中，该重复序列仅存在于非常高分子量的DNA中。
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