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首页> 外文期刊>Molecular and Cellular Biology >Identification of a nonhistone chromosomal protein associated with heterochromatin in Drosophila melanogaster and its gene.
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Identification of a nonhistone chromosomal protein associated with heterochromatin in Drosophila melanogaster and its gene.

机译：鉴定果蝇果蝇中异染色质的非组蛋白染色体蛋白及其基因。

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摘要

Monoclonal antibodies were prepared against a fraction of nuclear proteins of Drosophila melanogaster identified as tightly binding to DNA. Four of these antibodies were directed against a 19-kilodalton nuclear protein; immunofluorescence staining of the polytene chromosomes localized the antigen to the alpha, beta, and intercalary heterochromatic regions. Screening of a lambda gt11 cDNA expression library with one of the monoclonal antibodies identified a recombinant DNA phage clone that produced a fusion protein immunologically similar to the heterochromatin-associated protein. Polyclonal sera directed against the bacterial lacZ fusion protein recognized the same nuclear protein on Western blots. A full-length cDNA clone was isolated from a lambda gt10 library, and its DNA sequence was obtained. Analysis of the open reading frame revealed an 18,101-dalton protein encoded by this cDNA. Two overlapping genomic DNA clones were isolated from a Charon 4 library of D. melanogaster with the cDNA clone, and a restriction map was obtained. In situ hybridization with these probes indicated that the gene maps to a single chromosome location at 29A on the 2L chromosome. This general strategy should be effective for cloning the genes and identifying the genetic loci of chromosomal proteins which cannot be readily assayed by other means.
机译：制备了针对果蝇果蝇核蛋白中被鉴定为与DNA紧密结合的部分核蛋白的单克隆抗体。这些抗体中有四种针对19个千达尔顿核蛋白。多肽染色体的免疫荧光染色将抗原定位在α，β和cal间异色区域。用一种单克隆抗体筛选λgt11cDNA表达文库，鉴定出重组DNA噬菌体克隆，该克隆产生了与异染色质相关蛋白在免疫学上相似的融合蛋白。针对细菌lacZ融合蛋白的多克隆血清在Western印迹上识别出相同的核蛋白。从λgt10文库中分离出全长cDNA克隆，并获得其DNA序列。对开放阅读框的分析揭示了由该cDNA编码的18101道尔顿蛋白。从带有cDNA克隆的D. melanogaster的Charon 4文库中分离出两个重叠的基因组DNA克隆，并获得了限制性图谱。与这些探针的原位杂交表明该基因映射到2L染色体上29A处的单个染色体位置。这种一般策略对于克隆基因和鉴定染色体蛋白的遗传位点应该是有效的，而其他方法不能轻易地对其进行测定。

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