ISOLATION OF THE RHODOPSEUDOMONAS-SPHAEROIDES FORM I RIBULOSE 1,5-BIPHOSPHATE CARBOXYLASE OXYGENASE LARGE AND SMALL SUBUNIT GENES AND EXPRESSION OF THE ACTIVE HEXADECAMERIC ENZYME IN ESCHERICHIA-COLI
ISOLATION OF THE RHODOPSEUDOMONAS-SPHAEROIDES FORM I RIBULOSE 1,5-BIPHOSPHATE CARBOXYLASE OXYGENASE LARGE AND SMALL SUBUNIT GENES AND EXPRESSION OF THE ACTIVE HEXADECAMERIC ENZYME IN ESCHERICHIA-COLI
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DOI：
10.1016/0378-1119(86)90191-5
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发表时间：
1986-01-01
期刊：
影响因子：
3.5
通讯作者：
TABITA, FR
中科院分区：
文献类型：
--
作者：
GIBSON, JL;TABITA, FR
利用异源RuBPC/O探针，对克隆的球形红假单胞菌进行I型核酮糖1,5-二磷酸羧化酶/加氧酶（RuRPC/O）序列的集落杂交筛选。鉴定出了一种重组质粒，该质粒可与灰芽甘蓝和球芽甘蓝的II型RuBPC/O基因进行杂交。经聚丙烯酰胺凝胶电泳和Western免疫印迹分析，杂交的4kb SmaI片段在大肠杆菌中表达了与I型RuBPC/O相同的活性酶。
A library of cloned Rhodopseudomonas sphaeroides DNA was screened by colony hybridization for form I ribulose 1,5-bisphosphate carboxylase/oxygenase (RuRPC/O) sequences using heterologous RuBPC/O probes. A recombinant plasmid was identified that hybridized to both the Anacystis nidulans and the R. sphaeroides form II RuBPC/O genes. Subcloning of a hybridizing 4-kb SmaI fragment allowed expression of active enzymes in Escherichia coli that was identical to form I RuBPC/O based on polyacrylamide gel electrophoresis and Western immunoblot analysis.