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首页> 外文期刊>Journal of Biotechnology >Effect of the replication mode of a plasmid on the stability of multimericendoxylanase genes in Bacillus subtilis
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Effect of the replication mode of a plasmid on the stability of multimericendoxylanase genes in Bacillus subtilis

机译：质粒复制方式对枯草芽孢杆菌中多聚木聚糖酶基因稳定性的影响

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摘要

Effect of the replication mode of a plasmid on the stability of tandemly multimerized endoxylanase genes and a gene dose-dependent expression of the endoxylanase were studied in Bacillus subtilis. The structural genes encoding an endoxylanase, carrying its original promoter and ribosomal binding sequence, were tandemly multimerized and cloned into the Escherichia coli-B. subtilis shuttle plasmid, pJH27 Delta 88 or pMTL500(e), which has a rolling circle-replicon or a theta (theta)-replicon in B. subtilis, respectively. The cloned dimers in pJH27 Delta 88, which has a rolling circle-replicon, spontaneously rearranged to monomers in B. subtilis DB104, whereas those in pMTL500e, having a theta (theta)-replicon, were stably maintained. Expression level of the endoxylanase was proportional to the gene dosage in multimers. The endoxylanase activity in the supernatant increased from 80 U ml(-1) with pMTL-1x containing a monomer of the gene to 165 U ml(-1) with pMTL-4x containing a tetramer. These results indicate that high level expression of the endoxylanase gene can be obtained by tandemly multimerizing the genes in a plasmid with a theta (theta)-replicon.
机译：在枯草芽孢杆菌中研究了质粒复制模式对串联多聚内切木聚糖酶基因稳定性和内切木聚糖酶基因剂量依赖性表达的影响。编码带有内部启动子和核糖体结合序列的内切木聚糖酶的结构基因被串联多聚并克隆到大肠杆菌B中。枯草杆菌穿梭质粒pJH27 Delta 88或pMTL500（e），在枯草芽孢杆菌中分别具有滚动的圆形复制子或theta（theta）复制子。 pJH27 Delta 88中具有克隆的圆形复制子的克隆的二聚体可自发地重排至枯草芽孢杆菌DB104中的单体，而pMTL500e中具有theta（θ）复制子的二聚体则得以稳定地维持。内切木聚糖酶的表达水平与多聚体中的基因剂量成正比。上清液中的内切木聚糖酶活性从含该基因单体的pMTL-1x的80 U ml（-1）增加到含四聚体的pMTL-4x的165 U ml（-1）。这些结果表明，可以通过在具有θ-复制子的质粒中串联基因多聚化来获得内切木聚糖酶基因的高水平表达。

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