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球形红假单胞菌II型1,5-二磷酸核酮糖羧化酶/加氧酶基因在大肠杆菌中的克隆与表达

Cloning and expression in Escherichia coli of the form II ribulose 1,5-bisphosphate carboxylase/oxygenase gene from Rhodopseudomonas sphaeroides.

作者信息

Quivey R G, Tabita F R

出版信息

Gene. 1984 Nov;31(1-3):91-101. doi: 10.1016/0378-1119(84)90198-7.

Abstract

The gene encoding the form II ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBPC/O) from Rhodopseudomonas (R.) sphaeroides has been identified on a 3-kb EcoRI fragment and cloned into a broad-host-range, high-copy-number plasmid, using the gene from Rhodospirillum (Rs.) rubrum as a hybridization probe. Subclones of the gene from R. sphaeroides in pBR322 and pUC8 show substantial levels of expression and enzymatic activity in whole cells and crude cell extracts of Escherichia coli. This enzymatic activity has been shown to be similar in many respects to that of the protein purified from R. sphaeroides.

摘要

编码球形红假单胞菌(R. sphaeroides)II型1,5-二磷酸核酮糖羧化酶/加氧酶(RuBPC/O)的基因已在一段3kb的EcoRI片段上被鉴定出来,并以红螺菌(Rs. rubrum)的基因作为杂交探针,克隆到一个广宿主范围、高拷贝数的质粒中。来自球形红假单胞菌的该基因在pBR322和pUC8中的亚克隆在大肠杆菌的全细胞和粗细胞提取物中显示出大量的表达和酶活性。已证明这种酶活性在许多方面与从球形红假单胞菌纯化的蛋白质的酶活性相似。

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