Shechter I, Conrad D G, Hart I, Berger R C, McKenzie T L, Bleskan J, Patterson D
Eleanor Roosevelt Institute, Denver, Colorado 80206.
Genomics. 1994 Mar 1;20(1):116-8. doi: 10.1006/geno.1994.1135.
Recently, we reported the isolation of a cDNA encoding the human enzyme squalene synthase, the first step of sterol biosynthesis uniquely committed to synthesis of cholesterol (6). As such, it is likely that this enzyme occupies a critical regulatory position in the synthesis of cholesterol. As part of continuing studies of the role of this gene in cellular metabolism, we undertook the mapping of this gene on the human chromosomes. To localize the gene, we have first isolated a yeast artificial chromosome (YAC) containing the squalene synthase gene. We then used fluorescence in situ hybridization (FISH) with yeast DNA containing the YAC to localize the gene to chromosome 8. Assignment to human chromosome 8 was confirmed by polymerase chain reaction analysis of a somatic cell hybrid containing human chromosome 8. Use of a somatic cell hybrid regional mapping panel dividing chromosome 8 into several fragments localized the gene to 8p21-pter. Fractional length analysis of the FISH mapping placed the signal generated with this YAC at 8p22-p23.1.
最近，我们报道了编码人鲨烯合酶的cDNA的分离，鲨烯合酶是甾醇生物合成的第一步，专门负责胆固醇的合成（6）。因此，这种酶很可能在胆固醇合成中占据关键的调节位置。作为对该基因在细胞代谢中作用的持续研究的一部分，我们对该基因进行了人类染色体定位。为了定位该基因，我们首先分离出一个包含鲨烯合酶基因的酵母人工染色体（YAC）。然后我们使用含有该YAC的酵母DNA进行荧光原位杂交（FISH），将该基因定位到8号染色体。通过对含有8号人类染色体的体细胞杂种进行聚合酶链反应分析，证实了该基因定位于人类8号染色体。使用将8号染色体分成几个片段的体细胞杂种区域定位板，将该基因定位到8p21-pter。FISH定位的分数长度分析将该YAC产生的信号置于8p22-p23.1。
