摘要
为了提高瑞氏木霉（Trichoderma reesei）纤维素酶的活力,用类似基因改组的方法改造其碳源阻遏相关基因cre1。以瑞氏木霉基因组DNA为模板PCR扩增cre1基因,用DNaseⅠ消化cre1基因后,回收50-100 bp的DNA片段,用T4 DNA连接酶连接,以连接产物作为模板进行无引物PCR,并将PCR产物转入瑞氏木霉原生质体,通过测定滤纸酶活的方法筛选突变菌株,并在NCBI上比对分析突变菌株的cre1基因。结果表明,筛选获得1株纤维素滤纸酶活比出发菌株提高0.7倍的突变菌株cre2-3。cre2-3菌株在液体培养基中呈棉花状,而出发菌株呈小颗粒状,菌株cre2-3发酵液的颜色比出发菌株的更黄亮。推测cre1基因与瑞氏木霉菌株的生长代谢有关。
Cre1 gene mediating carbon catabolite repression was reformed by a method which was similar to DNA shuffling. The reconstructed cre1 gene fragment were transformed into T. reesei by protoplast transformation. Screening was carried out by measuring filter paper activities（FPA） of the culture supernatant. A stain named cre2-3was selected. Its filter paper activities was 0.7 times higher than that of original strain. There was obvious phenotypic difference between strain cre2-3and original strain.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2014年第2期288-292,共5页
Genomics and Applied Biology
基金
国家科技支撑课题(2011BAD22B01-01)
国家863项目(2012AA022106
2013AA050701)
国家国际合作项目(2010DFB63590
2011DFA61910)
广西科学研究与技术开发计划项目(桂科合10100019-21
桂科攻1099071
桂科合1140010-15)
广西自然科学基金项目(2012GXNSFAA053062)
广西科学院基本科研业务费资助项目(10YJ25SW15
11YJ24SW10)共同资助
关键词
碳源阻遏
DNA改组
cre1基因
瑞氏木霉
Carbon catabolite repression
DNA shuffling
cre1 gene
Trichoderma reesei
