根癌土壤杆菌介导棒形青霉的遗传转化

Genetic Transformation of Penicillium claviforme Mediated by Agrobacterium tumefacien

为实验室棒形青霉的功能基因研究奠定基础,采用根癌土壤杆菌（Agrobacterium tumefaciens）介导转化方法,以棒形青霉的分生孢子为受体,p DHt-sk-hyg为双元载体,成功实现棒形青霉的遗传转化。结果表明：棒形青霉对潮霉素B比较敏感,30μg/m L就可以抑制其生长;遗传转化的效率为70-90个转化子/106个分生孢子,连续转接5代能够稳定遗传;转化子的PCR检测表明,潮霉素磷酸转移酶标记基因已整合到棒形青霉的基因组中,转化子的遗传表现稳定。

In order to lay a foundation for studying on functional genes of P. claviforme in the laboratory,by using A. tumefaciens-mediated transformation,the authors successfully transformed P. claviforme and obtained T-DNA insertion mutants. Under the condition of p DHt-skhyg being binary vector,the transformation efficiency reached 70-90 transformants per 106 spores. Transformants were obtained under more than 30 μg / m L Hygromycin B. Most of them were quite stable after five rounds of successive cultures. PCR amplification showed that the TDNA was integrated into the genome,and was stable through mitotic cell division.