摘要:丝氨酸蛋白酶是一个蛋白酶家族，它们能断裂大分子蛋白质中的肽键，使之成为小分子蛋白质。本文对产丝氨酸蛋白酶菌株P. ananatis进行了丝氨酸蛋白克隆分析，结果表明：PCR产物长度为1062 bp,与其序列相似性最高的菌株是 Pantoea ananatis PA13(CP003085.1) ，相似度99.00%，翻译出其蛋白序列，长度为352个氨基酸，分子量是37013D。本研究有助于从分子生物学角度更深入地对产丝氨酸蛋白酶菌株进行分析利用。

关键词: 丝氨酸蛋白酶; Pantoea ananatis; 克隆

Abstract:The serine protease is a protease family, they are capable of cleaving the peptide bond in protein molecules, to become small molecule protein. Cloning and analysis of a serine protease from a protease -producing strain, P. ananatis, was carried out, and the results show, the length of the PCR product is 1062 bp, its highest similarity sequence is of the strain the Pantoea ananatis PA13 (CP003085.1), similarity is 99.00%. The present study also obtain its protein sequence with the length 352aa and 37013D . This study helps to have the characteristics of production serine protease strain from the perspective of molecular biology more in-depth analysis.

Keywords:  Serine protease，Pantoea ananatis，Cloning

目录

绪论-1

1.1蛋白酶的研究概况-1

1.1.1蛋白酶的研究背景-1

1.1.2蛋白酶的的种类及分布-1

1.1.3蛋白酶的应用及前景-1

1.2丝氨酸蛋白酶-2

1.2.1丝氨酸蛋白酶的分类-2

1.2.2丝氨酸蛋白酶的催化机制-3

1.2.3丝氨酸蛋白酶的应用-4

1.3丝氨酸蛋白酶相关基因的克隆-5

1.4 Pantoea ananatis的研究进展-5

1.5本研究的目的与意义-6

2 材料与方法-8

2.1 实验材料-8

2.1.1 实验菌株-8

2.1.2 主要药品-8

2.1.3 仪器设备-9

2.2培养基的配制与灭菌-10

2.2.1培养基的配制-10

2.3 丝氨酸蛋白酶基因克隆-11

2.3.1基因组DNA的提取-11

2.3.2 PCR扩增-12

2.3.3电泳检测-13

2.3.4胶回收PCR产物-14

2.3.5测序分析-15

3 结果与讨论-16

3.1实验结果-16

3.1.1丝氨酸蛋白酶基因的PCR扩增电泳结果-16

3.1.2 DNA序列测序结果-16

3.1.3丝氨酸蛋白酶的蛋白序列-17

3.2讨论-18

结论-20

致谢-21

参考文献-22