• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文会议>Journal of Anhui Agricultural University >cDNA Cloning an expression of Chitosanase from Aspergillus fumigatus
【24h】

cDNA Cloning an expression of Chitosanase from Aspergillus fumigatus

机译：烟曲霉壳聚糖酶表达的cDNA克隆

获取原文
获取原文并翻译 | 示例
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

An extracellular chitosannase was purified from Aspergillus fumigatus an N-terminal amino acids of the enzyme were sequenced. Oligonucleotide primer based on the sequence was used to PCR amplify fo the chitosanase cDNA. The cloned full length cDNA, 804 bp in size, encoded a single peptide of 251 amino acides. The blast search results ofthe cDNA sequence showed that the sequence shared th homology with chitosanase of Fusarium solani in two region, specifically but showed no homology with known bacterial chitosanases. The two region have repeated aspartic acid and threonin amino acid residue, respectively. This suggests that the chitosanase might have an evolutional origin distinct from bacterial chitosanase. SOuthern blot analysis results indicated that csn is present as a single copy in the genome of A.fumigatus. The recombinant protein corresponding to the mature protein was generated in E.coli using a plasmid vector pET28a(+). When csn cDNA corresponding to the mature protein was introduced into E.coli and expressed, approximately 28kDa protein was produced in a large amount after IPTG induction.
机译：从烟曲霉纯化细胞外壳聚糖酶，并对酶的N-末端氨基酸进行测序。基于该序列的寡核苷酸引物用于PCR扩增壳聚糖酶cDNA。克隆的全长cDNA（大小804 bp）编码了251个氨基酸的单个肽。 cDNA序列的blast搜索结果表明，该序列在两个区域与茄形镰刀菌的壳聚糖酶具有相同的同源性，但与已知的细菌壳聚糖酶没有同源性。这两个区域分别具有重复的天冬氨酸和苏氨酸氨基酸残基。这表明壳聚糖酶可能具有不同于细菌壳聚糖酶的进化起源。南方印迹分析结果表明，csn在烟曲霉基因组中以单拷贝形式存在。使用质粒载体pET28a（+）在大肠杆菌中生成与成熟蛋白相对应的重组蛋白。当将对应于成熟蛋白的csn cDNA导入大肠杆菌并表达时，IPTG诱导后大量产生约28kDa蛋白。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号