AN ACETOBACTER-XYLINUM INSERTION-SEQUENCE ELEMENT ASSOCIATED WITH INACTIVATION OF CELLULOSE PRODUCTION
AN ACETOBACTER-XYLINUM INSERTION-SEQUENCE ELEMENT ASSOCIATED WITH INACTIVATION OF CELLULOSE PRODUCTION
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DOI：
10.1128/jb.173.18.5723-5731.1991
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发表时间：
1991-09-01
影响因子：
3.2
通讯作者：
COUCHERON, DH
中科院分区：
文献类型：
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作者：
COUCHERON, DH
插入序列（IS）元件IS1031导致插入与木醋杆菌ATCC 23769的自发纤维素缺陷（Cel-）突变体相关。该元件是在木参ATCC 23769细胞突变体与木参NRCC 17005细胞突变体的原生质粒pAXC145杂交分析时发现的。在pAXC145上发现了一个IS元素IS1031B，显然与IS1031相同。IS1031大约是950 bp。DNA测序结果表明，这两个元件具有相同的末端，倒置重复24bp，包含两个错配，它们产生了3bp的目标序列重复。木香ATCC 23769野生型携带7个IS1031拷贝。南方杂交表明，ATCC 23769的17个独立分离的自发细胞突变体中有8个含有与IS1031同源的插入元件。大多数插入都在独特的位置，表明插入特异性较低。值得注意的是，两个插入位于最近发现的纤维素合酶基因上游0.5 kb处。试图在静态培养中通过选择分离这两种细胞插入突变体的自发产纤维素的复合体是不成功的。相反，获得了在液-气界面上形成蜡状膜的假可逆剂。这两个假回复性基因在基因组的不同位点上携带了一个is1031样元件的新插入，而没有切除先前的插入。综上所述，这些结果表明，土着IS元素有助于木质素的遗传不稳定性。这些元素也可能是这种生物和相关物种的有用遗传工具。
An insertion sequence (IS) element, IS1031, caused insertions associated with spontaneous cellulose deficient (Cel-) mutants of Acetobacter xylinum ATCC 23769. The element was discovered during hybridization analysis of DNAs from Cel- mutants of A. xylinum ATCC 23769 with pAXC145, an indigenous plasmid from a Cel- mutant of A. xylinum NRCC 17005. An IS element, IS1031B, apparently identical to IS1031, was identified on pAXC145. IS1031 is about 950 bp. DNA sequencing showed that the two elements had identical termini with inverted repeats of 24 bp containing two mismatches and that they generated 3-bp target sequence duplications. The A. xylinum ATCC 23769 wild type carries seven copies of IS1031. Southern hybridization showed that 8 of 17 independently isolated spontaneous Cel- mutants of ATCC 23769 contained insertions of an element homologous to IS1031. Most insertions were in unique sites, indicating low insertion specificity. Significantly, two insertions were 0.5 kb upstream of a recently identified cellulose synthase gene. Attempts to isolate spontaneous cellulose-producing revertants of these two Cel- insertion mutants by selection in static cultures were unsuccessful. Instead, pseudorevertants that made waxlike films in the liquid-air interface were obtained. The two pseudorevertants carried new insertions of an IS1031-like element in nonidentical sites of the genome without excision of the previous insertions. Taken together, these results suggest that indigenous IS elements contribute to genetic instability in A. xylinum. The elements might also be useful as genetic tools in this organism and related species.