抑制性消减杂交法筛选炭样小单孢菌JXNU-1中抗生素合成相关基因  

Screening of the Related-genes for Antibiotic Biosynthesis in Micromonospora carbonacea JXNU-1 by Suppression Subtractive Hybridization

作　　者：江云[1] 黄运红[1] 李非[1] 龙中儿[1] 

机构地区：[1]江西师范大学生命科学学院,南昌330022

出　　处：《基因组学与应用生物学》2016年第10期2733-2743,共11页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31360018);江西省自然科学基金项目(20132BAB204007;20161BAB204174)共同资助

摘　　要：为了筛选炭样小单孢菌JXNU-1中抗生素合成相关基因,本研究以培养36 h(抗生素分泌前)的炭样小单孢菌JXNU-1菌体所合成c DNA为Driver,以培养108 h(抗生素分泌后)的菌体所合成的c DNA为Tester,构建炭样小单孢菌JXNU-1分泌抗生素前后的差异c DNA消减文库,筛选差异表达基因,荧光定量PCR验证其表达量,生物信息学方法分析其功能。研究结果表明,经抑制性消减杂交筛选得到5个与抗生素合成相关的差异表达基因;比照炭样小单孢菌JXNU-1全基因组序列,将所获5个基因定位到4个生物合成基因簇中,最后确定了炭样小单孢菌JXNU-1中抗生素生物合成相关的基因簇。本研究为阐明炭样小单孢菌JXNU-1中抗生素的生物合成机制提供了实验依据。In order to screen the related-genes for antibiotic biosynthesis in Micromonospora carbonacea JXNU-1, the subtractive cDNA library of M. carbonacea JXNU- 1 was constructed with the cDNA from the mycelium of M. carbonacea JXNU-1 at the cultivation time of 108 h （antibiotics secretion） as tester and 36 h （non-secretion） as dr- iver respectively in this research, the differential expressed genes were screened and verified by real-time quantitative PCR, whose function were analyzed with bioinformatics methods. The results showed that 5 genes with differential expression were screened by suppression subtractive hybridization. Compared with the whole genome sequence ofM. carbonacea JXNU-1, 5 obtained genes were located into 4 biosynthetic gene clusters. And then the gene cluster closely related to biosynthesis of antibiotics in M. carbonacea JXNU-1 was determined. This research provided the experimental basis for revealing the mechanism of antibiotics biosynthesis in M. carbonacea JXNU-1.

关 键 词：炭样小单孢菌 抑制性消减杂交 抗生素合成相关基因 生物合成机制 

分 类 号：Q933[生物学—微生物学]