青风藤SaDBR2基因的克隆和表达.docxVIP

青风藤SaDBR2基因的克隆和表达

目录

一、内容概述................................................3

1.1研究背景与意义.......................................4

1.2研究目的与内容.......................................4

1.3技术路线与方法.......................................4

二、材料与方法..............................................5

2.1实验材料.............................................7

2.1.1青风藤组织样本...................................8

2.1.2基因克隆相关试剂.................................9

2.1.3转化与表达相关试剂...............................9

2.2实验方法............................................10

2.2.1样品制备........................................11

2.2.2基因克隆........................................12

2.2.3表达载体构建....................................12

2.2.4转化与表达......................................13

2.2.5结果检测........................................15

三、青风藤SaDBR2基因的克隆.................................16

3.1样品总RNA提取.......................................17

3.2cDNA第一链合成......................................17

3.3基因克隆............................................18

3.4验证克隆............................................19

四、青风藤SaDBR2基因的表达.................................20

4.1表达载体的构建......................................21

4.1.1启动子选择......................................21

4.1.2终止子选择......................................23

4.1.3其他元件添加....................................23

4.2转化与表达..........................................25

4.2.1转化方法........................................26

4.2.2表达载体转入青风藤细胞..........................27

4.3表达产物的检测......................................28

五、结果与分析.............................................29

5.1基因克隆结果........................................30

5.2表达载体构建结果....................................31

5.3转化与表达结果......................................32

5.4结果分析............................................33

六、讨论...................................................34

6.1基因克隆的策略与优化................................35