引用本文:	陈颖,李文彬,张利明,孙勇如.小球藻对5种常用基因工程抗生素的敏感性研究.海洋与湖沼,1999,30(5):500-505.

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小球藻对5种常用基因工程抗生素的敏感性研究
陈颖^1,2, 李文彬^1,2, 张利明^1,2, 孙勇如^1,2
1.中国科学院遗传研究所北京;2.100101

摘要:

于1996年以本实验室培养的椭圆小球藻为材料，就其对G418、潮霉素、氯霉素、链霉素、卡那霉素等5种抗生素的敏感性进行了研究。结果表明，小球藻对氯霉素、链霉素和卡那霉素不敏感，对潮霉素较为敏感，100 μg/ml（致死剂量，下同）即0.190 mmol/L（致死浓度，下同）的用量可抑制固体培养中小球藻的生长；对G418则高度敏感，30 μg/ml即0.043 mmol/L的G418即可完全抑制固体培养中小球藻的生长，15 μg/ml即0.022 mmol/L的G418即可完全抑制液体培养中小球藻的生长。与其它4种抗生素相比，G418更适合作为小球藻基因工程的选择试剂。

关键词: 小球藻基因工程选择试剂

DOI：

分类号:

基金项目:国家“九五”科技攻关资助项目，96-C02-04-05号

附件

STUDY ON SENSITIVITIES OF CHLORELLA ELLIPSOIDEA TO 5 ANTIBIOTICS IN GENETIC ENGINEERING

CHEN Ying,LI Wen-bin,Zhang Li-ming,SUN Yong-ru

Institute of Genetics, me Chinese Academy of Sciences, Beijing, 100101

Abstract:

Much progress has been made for recent years in selection markers of high plants. However, the situation varies for different algae. Researches on Chlamdomynas reinhardtii developed rapidly and there is also a good start in macroalgae, such as Laminaria japonica, but little progress has been made for Chlorella. Lack of selection marker made the use of gene transferred Chlorella limited. This paper reports a study on sensitivities of Chlorella ellipsoidea to 5 antibiotics in order to identify suitable selection marker of Chlorella genetic engineering. Chlorella ellipsoidea cells after 4–5 day’s culture were used as material and antibiotics—Hygromycin (Hm), Cholromycetin (Cm), and Strepeomycin (Str) and Kanamycin (Km); different concentrations were used for test in solid culture and Geneticin (G418) was used in solid and liquid culture. After 1–5 week’s culture, the results show that C. ellipsoidea is not sensitive to Cm, Str and Km; 600 μg/ml or 1.240 mmol/L Km, 600 μg/ml or 0.412 mmol/L Str and 200 μg/ml or 0.169 mmol/L Cm cannot inhabit the growth of C. ellipsoidea. However, it is relatively sensitive to Hm; 100 μg/ml or 0.190 mmol/L Hm can completely stop the growth of C. ellipsoidea on solid media. Further, C. ellipsoidea was highly sensitive to G418, since 30 μg/ml or 0.043 mmol/L and 15 μg/ml or 0.022 mmol/L completely inhabited the growth of C. ellipsoidea in solid and liquid cultures, respectively. It suggests that G418 be the most suitable selection regent if using NPTII gene as selection marker for C. ellipsoidea genetic engineering.

Key words: Chlorella ellipsoidea, Genetic engineering, Selection marker