根癌土壤杆菌介导棒形青霉的遗传转化
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。...
Saved in:
| Published in | 西南农业学报 Vol. 28; no. 2; pp. 700 - 703 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
贵州省农业科学院,贵州贵阳550006
2015
贵州大学生命科学学院,贵州贵阳,550025%贵州省农业生物技术重点实验室,贵州贵阳550006 贵州省生物技术研究所,贵州贵阳550006%贵州省农业生物技术重点实验室,贵州贵阳550006 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1001-4829 |
| DOI | 10.16213/j.cnki.scjas.2015.02.046 |
Cover
| Summary: | 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。 |
|---|---|
| Bibliography: | 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. SUN Chengdong, LIU Yong-xiang, PEN Xi-yi, WANG Yu-chen, LIU Zuo-yi. （ 1. College of Life Science, Guizhou University, Guizhcu Guiyang 550006, China ;2. Guizhcu Key Laboratory of Agricultural Biotechnology, Guizhou Guiyang 550006, China; 3. Guizhou Institute of Biotechnology, Guizhou Guiyang 550006, China; 4. Guizhou Academy of Agricultural Sciences, Guizhou Guiyang 550006, China） 51-1213/S Penicillium |
| ISSN: | 1001-4829 |
| DOI: | 10.16213/j.cnki.scjas.2015.02.046 |