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瑞氏木霉碳源阻遏相关基因Cre1的分子改造
引用本文:陈小玲,陈东,张穗生,陈英.瑞氏木霉碳源阻遏相关基因Cre1的分子改造[J].广西农业生物科学,2014(2):288-292.
作者姓名:陈小玲  陈东  张穗生  陈英
作者单位:广西科学院非粮生物质酶解国家重点实验室,国家非粮生物质能源工程技术研究中心,广西生物质炼制重点实验室,南宁530007
基金项目: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
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。

关 键 词:碳源阻遏  DNA改组  cre1基因  瑞氏木霉

Molecular Modification of Cre1 Gene Mediating Carbon Catabolite Repression in Trichoderma reesei
Chen Xiaoling,Chen Dong,Zhang Suisheng,Chen Ying.Molecular Modification of Cre1 Gene Mediating Carbon Catabolite Repression in Trichoderma reesei[J].Journal of Guangxi Agricultural and Biological Science,2014(2):288-292.
Authors:Chen Xiaoling  Chen Dong  Zhang Suisheng  Chen Ying
Institution:(State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-Food Biorefinery, Guangxi Key Laboratory of Biorefinery, Guangxi Academy of Science, Nanning, 530007)
Abstract: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.
Keywords:Carbon catabolite repression  DNA shuffling  cre1 gene  Trichoderma reesei
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