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δ-氨基乙酰丙酸脱水酶:人类基因的诱导表达及在染色体9q13至qter区域的定位

delta-Aminolevulinate dehydratase: induced expression and regional assignment of the human gene to chromosome 9q13----qter.

作者信息

Wang A L, Astrin K H, Anderson W F, Desnick R J

出版信息

Hum Genet. 1985;70(1):6-10. doi: 10.1007/BF00389449.

Abstract

The structural gene encoding human delta-aminolevulinate dehydratase has been assigned to the long arm of chromosome 9 by somatic cell hybridization techniques using murine erythroleukemia-human fibroblast somatic cell hybrids. Dimethyl sulfoxide induction of erythroid differentiation in these hybrid cells resulted in a 3 to 12-fold increase in the levels of total delta-aminolevulinate dehydratase. Human delta-aminolevulinate dehydratase was detected by an immunodiscrimination assay using polyclonal mouse anti-human delta-aminolevulinate dehydratase antibodies. Of four primary hybrid clones, each from an independent fusion, one hybrid line, XX-8, was positive for human delta-aminolevulinate dehydratase. Examination of 23 secondary, tertiary, and quaternary XX-8 subclones revealed that the expression of the human isozyme segregated with human chromosome 9q, confirming the provisional regional assignment made by classical linkage studies. One positive quaternary clone, XX-8-H21-H7-2, expressed human delta-aminolevulinate dehydratase activity and contained only human 9q13----qter. In addition, studies of tertiary and quaternary subclones from two series, XX-8-A31 and XX-8-H21-H7, indicated that murine regulatory factors increased the human as well as the murine enzymatic activity following induction of erythroid differentiation.

摘要

利用鼠类红白血病-人成纤维体细胞杂种的体细胞杂交技术,已将编码人δ-氨基-γ-酮戊酸脱水酶的结构基因定位于9号染色体长臂。这些杂种细胞中,二甲基亚砜诱导的红系分化使总δ-氨基-γ-酮戊酸脱水酶水平增加了3至12倍。使用多克隆小鼠抗人δ-氨基-γ-酮戊酸脱水酶抗体的免疫鉴别试验检测到人δ-氨基-γ-酮戊酸脱水酶。在四个独立融合产生的初级杂种克隆中,一个杂种系XX-8人δ-氨基-γ-酮戊酸脱水酶呈阳性。对23个XX-8二级、三级和四级亚克隆的检测表明,人同工酶的表达与人类9号染色体q区共分离,证实了经典连锁研究做出的初步区域定位。一个阳性四级克隆XX-8-H21-H7-2表达人δ-氨基-γ-酮戊酸脱水酶活性,且仅包含人类9q13----qter。此外,对来自两个系列XX-8-A31和XX-8-H21-H7的三级和四级亚克隆的研究表明,在诱导红系分化后,鼠类调节因子增加了人类以及鼠类的酶活性。

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