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荚膜红细菌C4-二羧酸转运基因的诱变、克隆及互补分析

Mutagenesis, cloning and complementation analysis of C4-dicarboxylate transport genes from Rhodobacter capsulatus.

作者信息

Hamblin M J, Shaw J G, Curson J P, Kelly D J

机构信息

Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, UK.

出版信息

Mol Microbiol. 1990 Sep;4(9):1567-74.

PMID:1962840
Abstract

Transposon mutagenesis was used to isolate insertion mutants of the photosynthetic bacterium Rhodobacter capsulatus which were unable to grow under aerobic conditions in the dark on malate, succinate or fumarate as sole carbon sources. Of five mutants isolated, all were deficient in C4-dicarboxylate transport. However, these mutants were still capable of photoheterotrophic growth, although at a slower rate than the wild type, on malate and succinate (but not fumarate). The mutated locus (designated dct) was complemented in trans using a cosmid gene bank. Subcloning and complementation analysis indicated that at least three closely linked genes essential for aerobic dicarboxylate transport were contained within an 8.3 kb region of the Rhodobacter capsulatus chromosome.

摘要

转座子诱变被用于分离光合细菌荚膜红细菌的插入突变体,这些突变体在黑暗有氧条件下,以苹果酸、琥珀酸或富马酸作为唯一碳源时无法生长。在分离出的五个突变体中,所有突变体在C4 - 二羧酸转运方面都存在缺陷。然而,这些突变体仍然能够进行光异养生长,尽管在以苹果酸和琥珀酸(但不是富马酸)为碳源时,其生长速度比野生型慢。使用黏粒基因文库对突变位点(命名为dct)进行反式互补。亚克隆和互补分析表明,荚膜红细菌染色体的一个8.3 kb区域内包含至少三个对有氧二羧酸转运至关重要的紧密连锁基因。

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