摘要
从红曲霉基因组中扩增出酸性蛋白酶基因Asp的编码区,构建其同源表达载体p BC-Hygro-Asp并导入到农杆菌GV3101备用。利用根癌农杆菌介导法将重组质粒导入红曲霉,并筛选获得Asp基因同源重组转化子,实现了酸性蛋白酶基因在红曲霉中的同源表达。转化子中酸性蛋白酶基因表达量是野生型菌株的3.30倍,能达到高产酸性蛋白酶的目的。对红曲霉酸性蛋白酶基因序列进行分析,结果显示该基因产物有两个天冬氨酸蛋白酶活性位点,为亲水性分泌蛋白,不参与信号转导,且与赤曲霉酸性蛋白酶有相同进化速率。
In this work,the coding region of Asp was amplified by PCR from the genomic DNA of the Monascus and the homologous vector pBC-Hygro-Asp was constructed and then was introduced into the Agrobactium tumefaciens strain GV3]0! For further study.The agrobacterial cells harboring the pBC-Hygro-Asp vector was introduced into the genome of Monascus via Agrobacterium tumefaciens-mediated method and the positive recombinant of Monascus was obtained.The expression level of acid protease gene in transformants strain was 3.30 times that of the wild Monascus, which indicated that the Monascus recombinant with high yield of acid protease was successfully achieved.The gene sequence of acid protease showed the gene had two aspartic proteases with active sites and the protease was a hydrophilic secretory protein without performance of signal transduction and keet the same evolution rate as that of Aspergillus ruber.
出处
《食品工业科技》
CAS
CSCD
北大核心
2016年第7期105-109,共5页
Science and Technology of Food Industry
基金
国家科技支撑计划(2014BAD07B02)
四川省科技支撑计划(2010NZ0093)
酿酒生物技术及应用四川省重点实验室项目(NJ2011-03
NJ2014-03)
重庆大学大型仪器设备开放基金
关键词
红曲霉
酸性蛋白酶
同源表达
序列分析
Monascus
acid protease
homologous expression
sequential analysis
