摘要
以抗生素抗性为选择标记进行角毛壳菌和球毛壳菌种间原生质体融合。角毛壳菌CH21－I－402菌株对潮霉素有抗性、对G418敏感，球毛壳菌CH08对潮霉素和G418敏感。根癌农杆菌介导的新霉素磷酸转移酶基因转化球毛壳菌，得到成功转化新霉素磷酸转移酶基因的球毛壳菌菌株，转化子对G418抗性提高3～4倍，对潮霉素仍然比较敏感。以耐潮霉素、不耐G418的角毛壳菌和不耐潮霉素、耐G418的球毛壳菌为出发菌株，以PEG6000为助融剂进行原生质体融合，用G418和潮霉素抗性进行筛选，获得再生菌株。经AFLP分析表明：再生菌株PF1、PF26为融合菌株，融合菌株菌落形态均与亲本存在明显差异。
A selection system for hybrids by protoplast fusion between Chaetomium cupreurn and C. globosum using dominant selective drug resistance markers were developed. C. cupreum was sensitive to G418 （Gs） and resistant to hygromycin B（H^T）. C. globosum was sensitive to both G418 （G^s） and hygromycin B （H^s ）. Agrobacterium tumefaciens mediated neomycin phosphotransferase II gene transformation of C. globosum （G^s, H^s） acquired resistance to G418 strains （G^r, H^s）. Protoplasts of C. cupreum （H^r, G^s） and C. globosum （H^s, G^r） were prepared, mixed in the presence of polyethylene glycol 6000 and incubated on regeneration agar plates. Two regenerated colonies showing Hr and Gr were regarded as hybrids by amplified fragment length polymorphisms analysis of the genomic DNA and they displayed a significant exchange of taxonomic characteristics between the parental strains.
出处
《华东理工大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第1期36-41,共6页
Journal of East China University of Science and Technology
基金
国家高技术研究发展计划(863计划)项目(2006AA10A213)
中国科学院重要方向资助项目(KSCX2-YW-G016)
关键词
角毛壳菌
球毛壳菌
抗生素抗性
原生质体融合
扩增片段长度多态性
Chaetomium cupreum
C. globosum
antibiotic resistance
protoplasts fusion
amplified fragment length polymorphism
