摘要
通过PCR的方法从苎麻脱胶菌株枯草芽孢杆菌B10中扩增出木聚糖酶基因。将该基因克隆在载体pB luescript II KS+上,并在该基因的前面连接CaMV 35S启动子,随后将潮霉素抗性基因(Hygr)片段克隆在上述载体上,成功构建了表达载体pKS-35SXYHyg。将该表达载体线性化后转化果胶酶产生菌黑曲霉菌株An1的原生质体。对抗潮霉素黑曲霉转化子进行基因组Southern杂交分析结果表明,木聚糖酶基因已整合到受体基因组中。与原出发菌株An1相比,黑曲霉转化子AT1的木聚糖酶活性提高2倍多,经AT1处理后苎麻的残胶率下降55.18%。
The xylanase gene of Bacillus subtilis B10 which had an excellent capacity for degumming ramie fibres was cloned by PCR amplification. The xylanase gene was cloned into vector pBluescript II KS+ and the CaMV 35S promoter was inserted in front of the gene, and the expression vector pKS-35SXYHyg was constructed successfully after the Hyg^(r) gene was inserted into the multi-cloning sites of pBluescript II KS+. The expression vector was linearized and transformed protoplasts of Aspergillus niger strain An1. Hygromycin-resistant transformants were generated and the integration of xylanase gene was confirmed by genomic Southern blotting analysis. Compared with An1, the xylanase activity of the transformant AT1 was increased more than trebled, and the residual gum content was decreased by 55.18% after the ramie was treated by culture supernatants from AT1.
出处
《微生物学通报》
CAS
CSCD
北大核心
2005年第3期62-67,共6页
Microbiology China
基金
重庆市应用基础研究项目(No.7975)
关键词
脱胶关键酶
基因克隆
黑曲霉
残胶率
Crucial degumming enzyme, Gene cloning, Aspergillus niger, Residual gum content
