摘要目的：研究小球藻对3种筛选用抗生素的敏感性，找到合适的抗生素来作为藻株改造的抗性标记基因。方法：在液体培养体系中筛选最适合用于小球藻筛选的抗生素。然后在液体培养基中加入不同浓度的抗生素，每天测定藻液的OD680值并计算抗生素对小球藻的生长抑制率(P％)。根据抗生素浓度(C)与抑制率(P％)的关系来评估小球藻对抗生素的敏感性。结果：经过1周的筛选试验，在液体培养基中，硫酸链霉素的致死剂量为100 µg/mL，氯霉素、氨苄青霉素钠的均无致死反应。在硫酸链霉素浓度为40 µg/mL时，颜色上明显看出小球藻的生长受到显著的抑制；当抗生素浓度达到100 µg/mL时已经完全抑制了小球藻的生长。因此，对于硫酸链霉素而言，100 µg/mL是小球藻生长的致死剂量。在液体正式培养中，硫酸链霉素的作用剂量与在预实验的基础上进行了细化，详细观察了各浓度硫酸链霉素影响下的小球藻生长状况。结论：小球藻对氯霉素、氨苄青霉素钠不敏感，对硫酸链霉素素表现出高度敏感性。与其它2种抗生素相比，硫酸链霉素更适合作为小球藻遗传改造的抗生素抗性选择标记。80808

毕业论文关键词：小球藻；抗生素；选择标记；基因工程；敏感性分析

Abstract Objective Our paper is to study the sensitivity of Chlorella vulgaris to 3 kinds of antibiotics and to find the appropriate antibiotics to transform the resistant marker genes。 Methods In order to screen the most suitable for the screening of antibiotics of Chlorella vulgaris in the liquid culture medium, we added different concentrations of antibiotics to the liquid medium。 Then we measured the algae liquid in OD680 every day and calculated the growth inhibition rate of antibiotics on Chlorella vulgaris (P%)。According to the antibiotic concentration (C) and the relationship between the inhibition rate (P%), we evaluate the sensitivity to antibiotics of Chlorella vulgaris。 Results After one week of screening test in the liquid culture medium, we find the lethal dose of streptomycin sulfate was 100 µg/mL。 There were no lethal reaction between chloramphenicol and ampicillin。 When thestreptomycin sulfate concentration was 40 µg/mL, Chlorella vulgaris growth was significantly inhibited。 When the mass concentration was 100 µg/mL, the growth of Chlorella vulgaris has been completely inhibited。 Therefore, for streptomycin sulfate, 100 µg/mL is a lethal dose of Chlorella vulgaris growth。 In liquid formal training, the concentration was refined on the basis of pre experiments。 We observed the growth of Chlorella vulgaris under different concentration of streptomycin sulfate carefully。 Conclusion Chlorella vulgaris is not sensitive to chloramphenicol and ampicillin, but streptomycin sulfate shows high sensitivity。 Compared with the other two kinds of antibiotics, Streptomycin sulfate is more suitable for antibiotic resistance markers in genetic transformation of Chlorella vulgaris as a selection marker。

Keywords:Chlorella vulgaris;Antibiotics;Selected marker;Gene engineering;Sensitivity analysis

目  录

第一章  绪论 1

1。1 选题的目的和意义 1

1。2 微藻 1

1。2。1 小球藻 2

1。2。2 营养盐和pH值对小球藻生长的影响 3

1。2。3 光照对小球藻生长的影响 4

1。3 抗生素及其分类 4

1。3。1 β-内酰胺类 5

1。3。2 氨基糖苷类 5

1。3。3 大环内酯类