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通过互补DNA原位杂交分析海德氏果蝇两个热休克基因座在组织培养细胞和唾液腺细胞中的活性。

The activity of two heat shock loci of Drosophila hydei in tissue culture cells and salivary gland cells as analyzed by in situ hybridization of complementary DNA.

作者信息

Sondermeijer P J, Lubsen N H

出版信息

Chromosoma. 1979 May 10;72(3):281-91. doi: 10.1007/BF00331090.

Abstract

Complementary DNA was made to poly A+ nuclear or polysomal RNA isolated from heat shock tissue culture cells of Drosophila hydei. A number of loci other than the four major heat shock loci are labelled after in situ hybridization of these cDNA preparations, while solution hybridization indicated that only about 10% of the cDNA was specific for heat shocked cells. Removal of the fraction of cDNA which could react with 25 degrees C RNA and subsequent in situ hybridization of heat shock specific cDNA indicated that locus 4-81 B, a major salivary gland heat shock locus, is also active at 25 degrees C in tissue culture cells, while locus 4-85 B is specifically activated by heat shock in tissue culture cells. This latter locus is not seen to be clearly puffed in salivary glands, but was shown to be active in that tissue both by direct autoradiography of salivary gland chromosomes after 3H-uridine labeling and by hybridization of cDNA to chromosomal RNA.

摘要

以从海德氏果蝇热休克组织培养细胞中分离的聚腺苷酸加核或多核糖体RNA合成互补DNA。这些cDNA制剂原位杂交后,除了四个主要热休克基因座外,还有许多其他基因座被标记,而溶液杂交表明只有约10%的cDNA对热休克细胞具有特异性。去除能与25℃RNA反应的cDNA部分,随后对热休克特异性cDNA进行原位杂交,结果表明,主要唾液腺热休克基因座4-81 B在组织培养细胞中于25℃时也有活性,而基因座4-85 B在组织培养细胞中被热休克特异性激活。后一个基因座在唾液腺中未观察到明显的胀泡现象,但通过3H-尿苷标记后唾液腺染色体的直接放射自显影以及cDNA与染色体RNA的杂交表明它在该组织中有活性。

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