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摇蚊 Glyptotendipes barbipes(施泰格)中的两种富含 AT 的卫星 DNA:在 Glyptotendipes barbipes 和 Chironomus thummi 的多线染色体中的分离与定位

Two AT-rich satellite DNAs in the chironomid Glyptotendipes barbipes (Staeger): isolation and localization in polytene chromosomes of G. barbipes and Chironomus thummi.

作者信息

Schmidt E R

出版信息

Chromosoma. 1980;79(3):315-28. doi: 10.1007/BF00327322.

Abstract

Two AT-rich satellite DNAs are present in the genome of Glyptotendipes barbipes. The two satellites have densities of 1,680 g/cm3 (=21% GC) and of 1.673 g/cm3 (=13% GC) in neutral CsCl-density gradients. The main band DNA has a density of 1.691 g/cm# (= 32% GC). This value is in agreement with the 33% GC=content of G. barbipes DNA calculated from thermal denaturation (TM=83 degrees C). - In brain DNA as well as in salivary gland DNA the two satellite sequences together comprise 12-15% of the total G. barbipes DNA. Comparisons of the density profiles of DNA extracted from polytene and non-polytene larval tissue gave no hints for under-replication of the satellite DNAs during polytenization. - The two satellite DNAs have been isolated from total DNA by Hoechst 33258-CsCl density centrifugation and then localized in the polytene salivary gland chromosomes by in situ hybridization. Both satellite sequences hybridize to all heterochromatic centromer bands of all four chromosomes of G. barbipes. Satellite I (1.673 g/cm3) hybridizes mainly with the middle of the heterochromatin, satellite II (1.680 g/cm3) hybridizes with two bands at the margin of the heterochromatin. In situ hybridization with polytene chromosomes of Chironomus thummi revealed the presence of G. barbipes satellite sequences also in the Ch. thummi genome at varios locations, mainly the centromere regions.

摘要

巴氏摇蚊(Glyptotendipes barbipes)的基因组中存在两种富含AT的卫星DNA。在中性CsCl密度梯度中,这两种卫星DNA的密度分别为1.680 g/cm³(=21% GC)和1.673 g/cm³(=13% GC)。主带DNA的密度为1.691 g/cm³(=32% GC)。该值与通过热变性(TM = 83℃)计算得出的巴氏摇蚊DNA的33% GC含量相符。——在脑DNA和唾液腺DNA中,这两种卫星序列共占巴氏摇蚊总DNA的12 - 15%。对从多线和非多线幼虫组织中提取的DNA密度图谱进行比较,未发现卫星DNA在多线化过程中存在复制不足的迹象。——通过Hoechst 33258 - CsCl密度离心从总DNA中分离出这两种卫星DNA,然后通过原位杂交将其定位在多线唾液腺染色体上。两种卫星序列均与巴氏摇蚊所有四条染色体的所有异染色质着丝粒带杂交。卫星I(1.673 g/cm³)主要与异染色质中部杂交,卫星II(1.680 g/cm³)与异染色质边缘的两条带杂交。对嗜菌摇蚊(Chironomus thummi)多线染色体进行原位杂交显示,巴氏摇蚊卫星序列在嗜菌摇蚊基因组的多个位置也有存在,主要在着丝粒区域。

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