根癌农杆菌介导工业化产甘油假丝酵母的遗传转化  

Genetic Transformation of Industrialized Strain Candida glycerinogenes by Agrobacterium tumefaciens

作　　者：饶志明[1] 张君胜[1] 沈微[1] 方慧英[1] 诸葛健[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室和工业微生物中心,江苏无锡214036

出　　处：《应用与环境生物学报》2007年第6期868-871,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金(No30570142);江苏省青年科技创新人才(学术带头人)基金(BK2006504);教育部长江学者和创新团队发展计划资助(IRT0532)资助~~

摘　　要：通过构建质粒pCAM3300-zeocin,将其电击转化到根癌农杆菌LBA4404中.将含有目的质粒pCAM3300-zeocin的根癌农杆菌和工业化产甘油假丝酵母(Candida glycerinogenes)WL2002-5共培养,利用zeocin抗性基因为筛选标记,实现了pCAM3300-zeocin对产甘油假丝酵母的转化.并根据产甘油假丝酵母的生长特性,对转化条件进行了优化,在共培养时间为24h,产甘油假丝酵母和根癌农杆菌的细胞比例为1∶(500～1000)时最高转化率达到2个转化子/104个酵母细胞.初步建立了根癌农杆菌介导转化产甘油假丝酵母的转化方法.In this research, the plasmid pCAM3300-zeocin was constructed and transformed into Agrobacterium tumefaciens. The A. tumefaciens containing the binary vector pCAM3300-zeocin was co-cultivated with industrialized strain Candida glycerologenesis WL2002-5. Then, transformants were screened on a plate containing 150 mg/L Zeocin. PCR results demonstrated that zeocin gene was integrated into nuclear genome of C. glycerologenesis. Research results indicated that zeocin gene could be utilized as selection marker and the Zeocin could be used as selection pressure in transformation of C. glycerologenesis. According to the growth characteristics of C. glycerologenesis, the transformation conditions were further optimized. The results showed that the transformation efficiency could reach two trasformants per 104 C. glycerologenesis cell when cocultivation time remained 24 hours and cell ratio of C. glycerologenesis and A. tumefaciens was 1：500 - 1000, The successfully construc- ted transformation system of C. glycerologenesis by A. tumefaciens can provide a powerful tool for further research of foreign genes expression in C. glycerologenesis.

关 键 词：产甘油假丝酵母 转化 根癌农杆菌 zeocin基因 

分 类 号：Q78[生物学—分子生物学]