热休克后表达受抑基因的mRNA差别显示和克隆

Differential Display and Cloning of a mRNA Species Whose Gene Expression Is Suppressed by Heat Shock

以人成骨肉瘤细胞株（ＨＯＳ－８６０３）作为热休克模型，用差别显示ｍＲＮＡ法（ｄｄｍＲＮＡ）筛选，发现和克隆了一个热休克后表达受抑的ｃＤＮＡ片段。在４３℃、４０分钟热处理后１小时，ＨＯＳ－８６０３细胞总ｃＤＮＡ图谱未见明显变化，但以该ｃＤＮＡ片段作为探针，ＲＮＡｄｏｔｂｌｏｔ和Ｎｏｒｔｈｅｒｎｂｌｏｔ证实，热休克后该基因的ｍＲＮＡ的量明显下降，而ｍＲＮＡ未见明显降解。我们将这个因热休克而选择性的表达受抑的基因命名为ＨＳＳＧ－１。对全长３１２ｂＰ的ｃＤＮＡ片段的序列分析表明，ＨＳＳＧ－１可能是一个未知的新基因。

A cDNA fragment, representing a heat shock-suppressed mRNA species, was differentially displayed and subsequently cloned from human osteogenic sarcoma cell line (HOS- 8603 ), which was used as a heatshock model. One hour after 43℃, 40min heat treatment, although there was no apparent change on the cellular total cDNA pattern of HOS-8603, the RNA dot blot and Northern blot proved, using the cloned cDNA fragment as probe, that the amount of the mRNA species was obviously decreasd without distinctive mRNA degradation. This reduced expression gene (HSSG ). The nucleotide sequence with the cDNA fragment of a size of 312bp indicated that the HSSG did not show sequence homology up to 65% to search in EMBL(data up to June, 1994 ).