摘要
[目的]研究福寿螺纤维素酶基因的原核表达。[方法]福寿螺(Ampullaria gigas)的纤维素酶基因Ce1Ag编码含395个氨基酸的多功能纤维素酶。利用RT-PCR方法技术,从福寿螺中克隆纤维素酶基因CelAg,并构建于表达载体pET-30a(+)上,再转化至大肠杆菌BL21(DE3),获得重组菌E.coliBDCelAg,进行发酵并分析纤维素酶的活性。[结果]可表达出有活性的纤维素酶,酶活可达8.31 U/ml。[结论]为纤维素酶基因今后的大规模生产研究奠定基础,并对纤维素的生物转化与利用的进一步研究和解决当前世界能源危机、粮食短缺和环境污染等问题具有重要的意义。
[ Objective ] The research aimed to study the expression of cellulose gene from Ampullaria gigas in Escherichia coll. [ Method ] The cellulase gene of Ampullaria gigas encoding 395 aminoins. CelAg gene being inserted in the expressing vector pET-30a（ ＋ ） was cloned by RT-PCR, then it was transformed into Escherichia coil BL2t （DE3）. The recombinant strain was fermented in shaken flask, and the activity of cellulase was analysised. [ Result ] The results indicated that the eellulase gene was expressed and the eellulase had high activity, which was up to 8.31 U/ml. [ Conclusion] The study can provide the basis for the production of cellulose gene in large scale in future, and has an important significance to deeply study the biological transformation and utilization of cellulose and to solve some questions, including world-wide energy crisis, food shortage and environmental pollution and so on.
出处
《安徽农业科学》
CAS
北大核心
2009年第19期8914-8915,8924,共3页
Journal of Anhui Agricultural Sciences
基金
重庆市科委自然科学基金(编号:8663)
关键词
福寿螺
纤维素酶
大肠杆菌
基因工程
Ampullaria gigas
CeUulase
Escherichia coli
Genetic engineering
