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一种具有适合报告基因系统特性的莱茵衣藻诱导型芳基硫酸酯酶。纯化、表征及分子克隆。

An inducible arylsulfatase of Volvox carteri with properties suitable for a reporter-gene system. Purification, characterization and molecular cloning.

作者信息

Hallmann A, Sumper M

机构信息

Lehrstuhl Biochemie I, Universität Regensburg, Germany.

出版信息

Eur J Biochem. 1994 Apr 1;221(1):143-50. doi: 10.1111/j.1432-1033.1994.tb18723.x.

Abstract

The multicellular green flagellate Volvox carteri synthesizes a periplasmic arylsulfatase in response to sulfur deprivation. The inducible enzyme has been purified to homogeneity and characterized. The corresponding gene and cDNA have been cloned. Determination of the sequence of genomic clones and comparisons to the cDNA sequence, revealed sixteen introns and seventeen exons that encode a 649-amino-acid polypeptide chain. Since the arylsulfatase enzyme is readily assayed using chromogenic substrates, but is not detectable in cells grown in sulfate-containing medium, the gene encoding arylsulfatase may be useful as a reporter gene in V. carteri. In addition, the highly regulated promoter of the arylsulfatase gene suggests its suitability as a tool for producing inducible expression vectors for cloned genes.

摘要

多细胞绿藻团藻(Volvox carteri)在硫缺乏时会合成一种周质芳基硫酸酯酶。这种诱导型酶已被纯化至同质并进行了特性鉴定。相应的基因和cDNA已被克隆。对基因组克隆序列的测定以及与cDNA序列的比较,揭示了16个内含子和17个外显子,它们编码一条649个氨基酸的多肽链。由于芳基硫酸酯酶很容易使用显色底物进行检测,但在含硫酸盐培养基中生长的细胞中无法检测到,因此编码芳基硫酸酯酶的基因可能作为团藻中的报告基因有用。此外,芳基硫酸酯酶基因高度调控的启动子表明它适合作为生产克隆基因诱导表达载体的工具。

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