[1]王娟,龚艳芬,来亚鹏,等.干扰MHR1对嗜热毁丝霉纤维素酶活性的影响[J].深圳大学学报理工版,2018,35(1):15-21.[doi:10.3724/SP.J.1249.2018.01015]
　WANG Juan,GONG Yanfen,LAI Yapeng,et al.The effect of RNA interference of MHR1 on cellulase activity in Myceliophthora thermophila[J].Journal of Shenzhen University Science and Engineering,2018,35(1):15-21.[doi:10.3724/SP.J.1249.2018.01015]
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干扰MHR1对嗜热毁丝霉纤维素酶活性的影响()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第35卷
期数:
2018年第1期
页码:
15-21
栏目:
生物工程
出版日期:
2018-01-12

文章信息/Info

Title:
The effect of RNA interference of MHR1 on cellulase activity in Myceliophthora thermophila
文章编号:
201801003
作者:
王娟1; 龚艳芬1; 2; 来亚鹏1; 邓婷婷1; 谢宁1; 刘刚1
1）深圳大学生命与海洋科学学院，深圳市微生物基因工程重点实验室，广东深圳 518060
2）深圳市海洋生物资源与生态环境重点实验室，广东深圳 518060
Author(s):
WANG Juan1;  GONG Yanfen1; 2;  LAI Yapeng1;  DENG Tingting1;  XIE Ning1;  and LIU Gang1
1) College of Life Sciences and Oceanography, Shenzhen Key Laboratory of Microbiology and Gene Engineering, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China
2) Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen 518060, Guangdong Province, P.R.China
关键词:
嗜热真菌; 嗜热毁丝霉; RNA干扰; 纤维素酶; MHR1因子; 生物质纤维素
Keywords:
thermophilic fungi;  Myceliophthora thermophila;  RNA interference;  cellulase;  MHR1 factor;  cellulose biomass
分类号:
Q 939.9
DOI:
10.3724/SP.J.1249.2018.01015
文献标志码:
A
摘要:
利用RNA-Seq技术，筛选到潜在的负调控因子 MHR1（fungal_TF_MHR1）． 为研究MHR1与纤维素酶基因表达之间的关联，设计基因mhr1的RNA干扰序列，并使用丙酮酸脱羧酶强启动子pdc成功构建mhr1基因的RNA干扰表达载体，经原生质体转化，获得一株干扰效率较高的阳性转化子MtR5．通过RT-qPCR、酶活测定和胞外蛋白浓度测定等分析发现，在诱导和非诱导培养条件下，转化子MtR5的胞外蛋白浓度、纤维素酶活性以及主要纤维素酶基因的表达量均高于野生型的．干扰转录因子MHR1可增加纤维素酶基因的表达量，研究表明，MHR1是一种与纤维素酶基因表达调控相关的转录因子．
Abstract:
The potential negative regulatory factor MHR1 was screened by RNA-Seq technique. A positive transformant MtR5 was successfully obtained by constructing the RNAi expression vector for mhr1 through the designing of the RNA interference sequence aimed at mhr1 and the use of the strong promoter of pdc gene. Real-time quantitative PCR, enzymatic and protein concentration determination show that the cellulase activities, extracellular protein concentration, and main cellulase genes expression of the transformant MtR5 are higher than those of the wild type under both inducing and non-inducing conditions. The above results elucidate the relationship between MHR1 and cellulase gene regulation for the first time and the association between the protein MHR1 and carbon catabolite repression. MHR1 works as a novel transcription factor associated with the cellulase gene expression and it is a repressor in carbon catabolite repression.

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备注/Memo

备注/Memo:
Received：2017-06-02；Accepted：2017-09-15
Foundation：National Natural Science Foundation of China (31601014) ; Shenzhen Science and Technology Research Foundation（JCYJ20150525092940997; JYJC20140418091413587）
Corresponding author：Professor LIU Gang. E-mail: zjuliug@szu.edu.cn
Citation：WANG Juan, GONG Yanfen, LAI Yapeng, et al. The effect of RNA interference of MHR1 on cellulase activity in Myceliophthora thermophila［J］. Journal of Shenzhen University Science and Engineering, 2018, 35(1): 15-21.（in Chinese）
基金项目：国家自然科学基金资助项目（31601014）；深圳市科技计划基础研究资助项目（JCYJ20150525092940997；JYJC20140418091413587）
作者简介：王娟（1975—），女，深圳大学副教授、博士．研究方向：应用微生物学. E-mail:wangjuan@szu.edu.cn
引文：王娟，龚艳芬，来亚鹏，等．干扰MHR1对嗜热毁丝霉纤维素酶活性的影响［J］. 深圳大学学报理工版，2018，35(1)：15-21.
更新日期/Last Update: 2017-12-22