1 绪论...... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ( 1)

1.1 何谓海洋生物技术........................... ................................................... ( 1)

1.2 海洋生物技术研究现状........................................................................ ( 2)

2 海洋微生物的筛选和培养........................................................................... ( 6)

2.1 概论... ... ... ...... ............... ... ... ......... ... ... ............ ... ... ... ............ ... ... ...... ( 6)

2.2 海洋微生物的採样、分离和培养技术...................................................... ( 7)

2.3 细菌的分类、鉴定与保存................................................................ . .(13)

2.4 产生EPA的海洋细菌的筛选............................................................... (19)

2.5 油细菌的筛选.................................................................................... (20)

2.6 海洋蓝藻的採集、分离和培养............................................................... (22)

3 海洋动物细胞的培养.................. ......... ... ... ... ...... ...... ... ... ... ... ...... ...... ... ... ... (28)

3.1 概论.................................................................. ......................... . .(28 )

3.2 鳗鲡、鲤鱼肝细胞的培养................................................................ . .(28)

3.3 鳗鲡肝细胞灌流法的操作................................................................ . .(29)

3.4 分离鳗鲡肝细胞的原代单层培养法......................................................... (34)

3.5 振荡法分离鲤鱼肝细胞................................................................... . .(35)

3.6 分离鲤鱼肝细胞的原代单层培养............................................................ (35)

3.7 培养肝细胞的实验........................ .................. ............................... . .(3 6)

3.8 草鱼肾组织细胞的原代培养和传代培养实验............................................. (39)

4 大型海藻的组织培养............................................................................ . .(4 1)

4 .1 概论. . ... ......... ... ... ... ... ......... ... ... ... ... ... ... ... ... ... .. ... ... ... ... ... ... ... . . .(41 )

4.2 组织培养的设备和基本技术............................................................. . .(43)

4.3 外植体的无菌化技术...................................................................... . .(48)

4.4 海藻组织块愈伤组织的诱导............................................. .................... (49)

4.5 无菌培养系的愈伤组织诱导.................................................................. (50)

4.6 细胞培养.......................................................................................... (51)

5 海洋动物的细胞操作........................ ......................................................... (56)

5.1 概论... ................................................... .......................................... (56)

5.2 性决定(性别控制)..................... ......................................................... (59)

5.3 嵌合体动物的培育.............................. ................................................ (62)

6 大型海藻的细胞操作............ ... ... ... ......... ... ... ............................ ... ......... ... ... (65)

6.1 概论................................................................................................ (65)

6.2 大型海藻的培养................................................................................. (68)

6.3 原生质体分离技术......................................................................... . .(70)

6.4 原生质体培养技术......................................................................... . .(74)

6.5 细胞融合技术............................................................................... . .(77)

6.6 异种融合细胞的筛选及个体再生............................................................ (82)

7 海洋微藻的细胞操作... ... ............... ... ... ......... ... ... ............ ... ... ............ ......... (85)

7.1 概论............................................. ........................... ... ..................... (85)

7.2 海洋微藻的分离和纯化技术.................................................................. (88)

7.3 培养基.................................... ......................................................... (91)

7.4 海洋微藻的生长测量和培养技术............................................................ (96)

7.5 原生质体分离技术........................................................................... (100)

7.6 原生质体培养技术.................. ......................................................... (103)

7.7 细胞融合技术................................................................................. (104)

7.8 细胞学技术................................................................................ .(105)

8 海产鱼类的染色体操作........................................................................... (109)

8.1 概论... ................................................... ....................................... (109)

8.2 天然多倍体......... ...... ............ ... ......... ... ... ... ... ...... ... ... ... ............ ...... (109)

8.3 人工诱导的多倍体...... ..................................................................... (111)

8.4 单性发育................................................................................... .(118)

8.5 染色体片断移植.............................................................................. (123)

8.6 染色体操体技术的套用及其发展趋势......................................... . . . .(123)

9 海产无脊椎动物的染色体操作......... ......................................................... (129)

9.1 概论... ............ ... ... ............ ...... ...... ... ... ... ......... ... ... ............ ... ......... (129)

9.2 细胞核及染色体的观察方法............................................................... (132)

9.3 人工多倍体诱导法........................................................................... (135)

9.4 多倍体的鉴定................................................................................. (137)

10 海洋动物的基因克隆.............................................................................. (141)

10.1 概论... ... ... ... ... ... ... ... ...... ... ... ... ......... ... ... ... ... ... ... ......... ... ... ...... ...... (141)

10.2 海洋动物的cDNA 克隆.................................................................. (143)

10.3 目的cDNA克隆的鉴定.................................................................. (166)

10.4 DNA 序列测定.............................................................................. (181)

11 转基因海洋动物........................ ............................................................ (190)

11.1 概论..................................................................... ........................ (190 )

11.2 基因导入方法................................................................................. (195)

11.3 转基因动物的鉴别...................................................................... . . (198)

12 海洋微藻的基因工程...... ........................................................................ (200)

12.1 概论......... ... ... ... ...... ...... ... ... ............ ... ... ............ ... ... ... ............... ... (200)

12.2 载体的开发............................................................... ..................... (201)

12.3 受体系统.......................................................... ....................... . . (202)

12.4 外源基因转移方法........................................................................... (203)

13 大型海藻的基因工程................................. ......... . . ............................. (208)

13.1 概论............................................................................................. (208)

13.2 材料的选择.................................................................................... (210)

13.3 大型海藻的核酸製备............................................. ........................... (210)

13.4 大型海藻的目的基因................................................................... . . (211)

14 纯种保存... ... ... ... ... ... ... ... ... ... ... ...... ... ......... ... ... ... ... ... ... ... ... ... ... ... ... ... ... (213)

14.1 概论............................................................................................. (213)

14.2 大型藻类的纯种保存........................................................................ (215)

14.3 海洋微型生物的冷冻保存.................................................................. (217)

14.4 海洋植物的冷冻保存........................................................................ (220)

14.5 海洋动物生殖细胞的冷冻保存............................................................ (222)

15 生物反应器.......................................................................................... (225)

15.1 概论............................................................................................. (225)

15.2 微生物固定化......... ............................................................ ............ (226)

15.3 利用生物反应器生产有用物质............................................................ (228)

15.4 海洋蓝藻生物反应器........................................................................ (231)

16 生物感测器... ... ......... ... ... ... ... ......... ... ... .................. ............ ... ... ... ......... (235)

16.1 概论............................................................................................. (235)

16.2 接受器的固定化.............................................................................. (237)

16.3 换能器.......................................................................................... (239)

16 .4 套用... . .. . .. .. . . . . . . . .. . .. . . . .. . .. . . .. . .. .. . . .. . .. . . . . . . .. . .. . . .. . . . .. . . .. . . . .. . . .. . . . (24 1)

16.5 结语.............................................................................. ............... (243)

17 海洋环境保护............................................................... ........................ (245)

17.1 概论............................................................................................. (245)

17.2 石油的生物降解.............................................................................. (247)

17.3 卤代烃的生物降解........................................................................... (248)

17.4 酶工程方法消除工业废水中的酚与芳香胺............................................. (250)

17.5 藻类对重金属离子的吸附.................................................................. (253)

17.6 可降解塑胶................................................................................ .(255)

17.7 生物技术在解决生物耗氧化合物对环境污染方面的套用........................... (256)

18 疾病的诊断与防治................................................................................. (257)

18.1 概论............................................................................................. (257)

18.2 诊断........................... .................................................................. (258)

18.3 防治............................................................................................. (260)

19 海洋生物活性物质... ... ... ... ... ... ... ... ... ...... ... ...... ... ...... ... ... ... ... ... ... ...... ... ... (261)

19. 1 概论... ... ... ... ... ... ... ... ... ... .... . ....... ... ... ... ... ...... ... ... ... ... ... ... ...... .. .(261)

19.2 海洋生物活性物质的研究方法........................................................ .(267)

20 海洋新材料— —甲壳质....................................................................... .(271)

20.1 概论... ... ...... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ...... ... ... ... ... ... ... ... ... ... ... ... (271)

20.2 人工皮肤............ ........................................................................... (272)