小球藻基因工程选择标记研究

Study of selectable markers in genetic engineering of Chlorella sp.

研究了小球藻对10种常见抗生素:庆大霉素、新霉素、氨苄青霉素、卡那霉素、头孢霉素、链霉素、G418、潮霉素、Zeocin和氯霉素的敏感性,发现小球藻对庆大霉素、新霉素、氨苄青霉素、卡那霉素、头孢霉素和链霉素不敏感;对G418和潮霉素较敏感;对Zeocin和氯霉素最敏感.利用统计学的方法和原理,确定了G418、潮霉素、Zeocin和氯霉素在海水培养基和淡化10倍的培养基中对小球藻的半致死剂量和95%可信限.为筛选出小球藻基因工程藻株的选择试剂,建立其遗传转化系统奠定了基础.

The sensitivities of Chlorella sp. to 10 antibiotics： gentamicin, neomycin, ampicillin, kanamycin, cephalothin, streptomycin, G418, hygromycin Zeocin and chloramphenicol were investigated. The results show that Chlorella sp. is not sensitive to gentamicin, neomycin, ampicillin, kanamycin, cephalothin and streptomycin, and they cannot inhibit the growth of Chlorella sp. in the concentration of this test. However, it is relatively sensitive to G418 and hygromycin, and very sensitive to Zeocin and chloramphenicol. LD50 and 95% confidence ranges of these four reagents in both seawater culture and decreasing NaCl concentration medium were calculated respectively. The results provide important data in developing selectable markers in genetic engineering of Chlorella sp.