Kondo K, Horinouchi S
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan.
Appl Environ Microbiol. 1997 Mar;63(3):1139-42. doi: 10.1128/aem.63.3.1139-1142.1997.
A novel insertion sequence element, IS12528, was found to be associated with inactivation of the alcohol dehydrogenase by insertion in the adhA gene, which encodes the primary dehydrogenase subunit of the three-component membrane-bound alcohol dehydrogenase complex in Gluconobacter suboxydans. Cloning and sequencing analyses revealed that IS12528 was 905 bp in length and had a terminal inverted repeat of 18 bp. In addition, IS12528 was found to generate a 3-bp duplication (TMA, where M represents C or A) at the inserted site upon transposition. IS12528 encoded one long product of 274 amino acids that was rich in basic amino acids. This protein showed significant homology with putative transposases of the IS1031 family isolated from Acetobacter xylinum, which belongs to another genus of acetic acid bacteria. IS12528-like sequences were distributed in a wide variety of acetic acid bacteria, as determined by Southern hybridization and PCR. These observations suggest that IS12528 is one of the insertion sequences that are responsible for genetic instability leading to deficiencies in various physiological properties in a variety of acetic acid bacteria.
发现一种新型插入序列元件IS12528与乙醇脱氢酶失活有关,它通过插入adhA基因实现,该基因编码氧化葡萄糖酸杆菌中三组分膜结合乙醇脱氢酶复合物的主要脱氢酶亚基。克隆和测序分析表明,IS12528长度为905 bp,具有18 bp的末端反向重复序列。此外,发现IS12528在转座时会在插入位点产生一个3 bp的重复序列(TMA,其中M代表C或A)。IS12528编码一个由274个氨基酸组成的长产物,该产物富含碱性氨基酸。该蛋白质与从木醋杆菌(属于另一属醋酸菌)中分离出的IS1031家族推定转座酶具有显著同源性。通过Southern杂交和PCR确定,类似IS12528的序列分布在多种醋酸菌中。这些观察结果表明,IS12528是导致多种醋酸菌各种生理特性缺陷的遗传不稳定的插入序列之一。