REMI介导电转化产甘油假丝酵母  

Genetic Transformation of Candida glycerinogenes by REMI and Electroporation

作　　者：张永光[1] 沈微[1] 饶志明[1] 方慧英[1] 诸葛健[1] 

机构地区：[1]江南大学生物工程学院工业微生物研究中心工业生物技术教育部重点实验室,无锡214036

出　　处：《微生物学通报》2007年第5期848-851,共4页Microbiology China

基　　金：江苏省青年科技创新基金资助项目(NoBK2006504);国家自然科学基金资助项目(No30570142)

摘　　要：为了分离耐高渗和甘油代谢相关基因,以Zeocin为选择标记,利用REMI技术电转化产甘油假丝酵母Candida glycerinogenes。考察了7种限制性内切酶对转化的影响,选择HindIII进一步优化了转化的几个条件。结果表明,在OD600≈1.3时收集细胞,在1.5kV电压下,感受态细胞浓度为2.0×109个细胞/mL,100U Hind III时,能获得129个转化子/μgDNA的较高转化率,58%的转化子稳定,表明REMI技术适合于产甘油假丝酵母的转化。In order to isolate genes related with the osmoadaptation and glycerol metabolism of Candida glycerinogenes, a transformation system based on the dominant selectable marker Zeocin and restriction enzyme-mediated integration （REMI） was established. Effects of seven restriction enzymes on transformation efficiency of C. glycerinogenes were tested. Transformation conditions were optimized in the presence of Hind Ill. Under the optimal conditions of OD600≈ 1.3, voltage of 1.5 kV, 2.0 × 10^9 competent cens/mL, 100 units of Hind III added, the transformation efficiency was up to 129 trnaformants/μg DNA. 58% of transformants were stable on nonselective medium. These results suggest that REMI technique would be beneficial to the genetic transformation of C. glycerinogenes.

关 键 词：产甘油假丝酵母 遗传转化 REMI 

分 类 号：Q78[生物学—分子生物学]