摘要:絲氨酸蛋白酶是一個(gè)蛋白酶家族，它們能斷裂大分子蛋白質(zhì)中的肽鍵，使之成為小分子蛋白質(zhì)。本文對產(chǎn)絲氨酸蛋白酶菌株P(guān). ananatis進(jìn)行了絲氨酸蛋白克隆分析，結(jié)果表明：PCR產(chǎn)物長度為1062 bp,與其序列相似性最高的菌株是 Pantoea ananatis PA13(CP003085.1) ，相似度99.00%，翻譯出其蛋白序列，長度為352個(gè)氨基酸，分子量是37013D。本研究有助于從分子生物學(xué)角度更深入地對產(chǎn)絲氨酸蛋白酶菌株進(jìn)行分析利用。

關(guān)鍵詞: 絲氨酸蛋白酶; 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

目錄

緒論-1

1.1蛋白酶的研究概況-1

1.1.1蛋白酶的研究背景-1

1.1.2蛋白酶的的種類及分布-1

1.1.3蛋白酶的應(yīng)用及前景-1

1.2絲氨酸蛋白酶-2

1.2.1絲氨酸蛋白酶的分類-2

1.2.2絲氨酸蛋白酶的催化機(jī)制-3

1.2.3絲氨酸蛋白酶的應(yīng)用-4

1.3絲氨酸蛋白酶相關(guān)基因的克隆-5

1.4 Pantoea ananatis的研究進(jìn)展-5

1.5本研究的目的與意義-6

2 材料與方法-8

2.1 實(shí)驗(yàn)材料-8

2.1.1 實(shí)驗(yàn)菌株-8

2.1.2 主要藥品-8

2.1.3 儀器設(shè)備-9

2.2培養(yǎng)基的配制與滅菌-10

2.2.1培養(yǎng)基的配制-10

2.3 絲氨酸蛋白酶基因克隆-11

2.3.1基因組DNA的提取-11

2.3.2 PCR擴(kuò)增-12

2.3.3電泳檢測-13

2.3.4膠回收PCR產(chǎn)物-14

2.3.5測序分析-15

3 結(jié)果與討論-16

3.1實(shí)驗(yàn)結(jié)果-16

3.1.1絲氨酸蛋白酶基因的PCR擴(kuò)增電泳結(jié)果-16

3.1.2 DNA序列測序結(jié)果-16

3.1.3絲氨酸蛋白酶的蛋白序列-17

3.2討論-18

結(jié)論-20

致謝-21

參考文獻(xiàn)-22