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苜蓿根瘤菌41中的聚-β-羟基丁酸酯(PHB)生物合成基因。

Poly-beta-hydroxybutyrate (PHB) biosynthetic genes in Rhizobium meliloti 41.

作者信息

Tombolini R, Povolo S, Buson A, Squartini A, Nuti M P

机构信息

CRIBI Biotechnology Center, Università di Padova, Italy.

出版信息

Microbiology (Reading). 1995 Oct;141 ( Pt 10):2553-9. doi: 10.1099/13500872-141-10-2553.

Abstract

Genes encoding beta-ketothiolase (phaA), acetoacetyl-CoA reductase (phaB) and PHB-synthase (phaC) from R. meliloti 41, together with a fourth gene, referred to as ORF1, presumed to be involved in PHB biosynthesis, have been cloned and sequenced. phaA, phaB and ORF1 were identified by heterologous hybridization on a cosmid library, while phaC was isolated by cloning the transposon-tagged fragment from a R. meliloti PHB- Tn5 mutant. phaA and phaB were functionally expressed in Escherichia coli while phaC was able to complement a PHB- strain of R. meliloti 41. The three genes were sufficient to direct the production of polyhydroxyalkanoate in E. coli. The homology of ORF1 with an ORF located near the PHB genes in two phototrophic bacteria suggests its involvement in PHB synthesis.

摘要

编码来自苜蓿中华根瘤菌41的β-酮硫解酶(phaA)、乙酰乙酰辅酶A还原酶(phaB)和聚羟基丁酸酯合酶(phaC)的基因,连同第四个基因(称为ORF1,推测参与聚羟基丁酸酯生物合成)已被克隆和测序。phaA、phaB和ORF1通过在黏粒文库上的异源杂交鉴定,而phaC是通过从苜蓿中华根瘤菌聚羟基丁酸酯-Tn5突变体中克隆转座子标签片段分离得到的。phaA和phaB在大肠杆菌中功能性表达,而phaC能够互补苜蓿中华根瘤菌41的聚羟基丁酸酯缺陷菌株。这三个基因足以指导大肠杆菌中聚羟基链烷酸酯的产生。ORF1与两种光合细菌中位于聚羟基丁酸酯基因附近的一个开放阅读框的同源性表明其参与聚羟基丁酸酯合成。

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