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[果蝇间带多线染色体插入的细胞遗传学分析]

[Cytogenetic analysis of insertions into drosophila interband polytene chromosomes].

作者信息

Semeshin V F, Chernukhin V A, Shabel'nikov I V, Omel'ianchuk L V, Beliaeva E S, Zhimulev I F

出版信息

Genetika. 1994 Jul;30(7):927-33.

PMID:7958809
Abstract

Using the method of P-element-mediated enhancer detection, 29 Drosophila melanogaster lines were obtained that carried P-1ArB vector insertions in chromosomes 2 and 3. The expression of the reporter gene lacZ at different developmental stages of the transformed lines was determined in color reactions for beta-galactosidase. Regions of vector integration were located using in situ hybridization. Subsequent electron-microscopic mapping of the transformed regions was performed using four lines (nos. 12, 41, 2, and 3). In the lines 12 and 41, lacZ was expressed in most tissues of embryos, larvae, and imagoes (including salivary glands), whereas in lines 2 and 3 its expression was observed only in embryos. Lines 12 and 2 showed the presence of insertions in the 85D9/10 and 86B4/6 interband regions, respectively. The absence of a novel band in line 3 could be associated with transposon integration into the band. In line 41, puffing of the transformed region was observed, which did not allow us to determine the presence of any novel structures in it. The novel structures in lines 12 and 2 looked like single bands with a similar DNA packing ratio of about 30. These bands were obviously polygenic, because the inserted vector contained four functionally different genes.

摘要

利用P因子介导的增强子检测方法,获得了29个黑腹果蝇品系,这些品系在2号和3号染色体上携带P-1ArB载体插入片段。通过β-半乳糖苷酶的显色反应,测定了转化品系在不同发育阶段报告基因lacZ的表达情况。利用原位杂交确定载体整合区域。随后,使用4个品系(第12、41、2和3号)对转化区域进行了电子显微镜定位。在第12和41号品系中,lacZ在胚胎、幼虫和成虫的大多数组织(包括唾液腺)中表达,而在第2和3号品系中,仅在胚胎中观察到其表达。第12和2号品系分别在85D9/10和86B4/6带间区域存在插入。第3号品系中没有新带可能与转座子整合到带中有关。在第41号品系中,观察到转化区域出现胀泡,这使得我们无法确定其中是否存在任何新结构。第12和2号品系中的新结构看起来像单带,其DNA包装比相似,约为30。这些带显然是多基因的,因为插入的载体包含四个功能不同的基因。

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