| 摘要: |
| 于1997年1－4月，以本所培养的孤雌生殖海带为材料，运用统计学方法研究其对氯霉素、潮霉素的敏感性，以期得出氯霉素、潮霉素对不同长度孤雌生殖海带的半致死剂量。结果表明，孤雌生殖海带长度（全长）在0.5－1.5cm时，氯霉素的半致死剂量与海带的长度相关，而潮霉素的半致死剂量与长度不相关。孤雌生殖海带对潮霉素比对氯霉素更敏感，提示潮霉素磷酸转移酶基因有可能成为海带基因工程的另一个选择标记。 |
| 关键词: 海带 氯霉素 潮霉素 选择标记 |
| DOI： |
| 分类号: |
| 基金项目:国家攀登计划B资助项目，PD B-6-4-1号;国家自然科学基金资助项目，39400076号 |
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| SENSITIVITY OF SPOROPHYTES OF PARTTHENOGENETIC LAMINARIA JAPONICA TO CHLORAMPHENICOL AND HYGROMYCIN |
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LI Xin-ping, QIN Song, ZENG Cheng-Kui (C.K. Tseng)
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Institute of Oceanology, ne Chinese Acadeny of Sciences, Qingdao, 266071
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| Abstract: |
| In order to optimize the selection condition for transgenic Laminaria japonica, the sensitivity of sporophytes of partthenogenetic L. japonica to Chloramphenicol and Hygromycin was tested from Jan. to Apr., 1997. Young sporophytes of partthenogenetic L. japonica were regenerated from partthenogenetic gametophytes and cultured in laboratory. The total lengths of young sporophytes were from 0.5 to 1.5cm. In the test for Chloramphenicol, the young sporophytes were divided into three groups according to their total lengths: a) (4.9±0.6)mm, b) (6.5±0.5)mm, and c) (8.4±0.9)mm. Chloramphenicol concentration in each group was 0, 50.0, 65.0, 84.5, 110.0, 143.0, 185.5, and 241.2μg/ml, respectively. In the test for hygromycin, the young sporophytes were also divided into three groups i.e. a) (6.3±1.0)mm, b) (8.3±1.0)mm, and c) (12.3±2.3)mm. Hygromycin concentration in each group were 0, 50.0, 65.0, 84.5, 110.0, and 143.0μg/ml, respectively. Steril polystyrene dishes (diameter 3.0cm) were used, which contains 5ml enriched seawater added with necessary antibiotics stock. Every ten young sporophytes were put into a plate, cultured at (10.0±0.5) °C in about 50μE/(m2*s), 10h light/14h darkness, and their death were observed every day. All media were renewed every week.
LD50 of Chloramphenicol at 15, 16, 17, 18, 19 and 20d and LD50 of hygromycin at 36, 48, 72 and 96h were calculated. The results show that the LD50 of L. japonica to chloramphenicol is correlated with total length (0.5-1.5cm), but the LD50 of hygromycin is not. The parthenogenetic sporophytes are more sensitive to hygromycin than to chloramphenicol. For example, LD50 of chloramphenicol in the three groups at 15d are 158.9, 110.6 and 89.0μg/ml, respectively, but LD50 of hygromycin in the three groups at 96h (4d) are only 47.4, 50.7 and 69.4μg/ml, respectively. This observation suggests that HPH (hygromycin phosphotransferase) gene is a more efficient selectable marker than CAT(chloramphenicol acetyltransferase) gene. |
| Key words: Laminaria japonica, Chloramphenicol, Hygromycin, Selectable marker |
