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Gene of internal cutting glucanase of Bacillus megatherium and preparation process thereof

A technology of Bacillus megaterium and endoglucanase, which is applied in the field of Bacillus megaterium endoglucanase gene and its preparation, and can solve the problem of no discovery of Bacillus megaterium glucanase coding gene fragment report and the like

Inactive Publication Date: 2007-03-21
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the query results in GenBank and various biological journals, no report on the gene fragment encoding Bacillus megaterium glucanase was found

Method used

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  • Gene of internal cutting glucanase of Bacillus megatherium and preparation process thereof
  • Gene of internal cutting glucanase of Bacillus megatherium and preparation process thereof
  • Gene of internal cutting glucanase of Bacillus megatherium and preparation process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] The sequence of the Bacillus megaterium endoglucanase coding gene fragment is as follows:

[0088] ggctgcgcaa ctgttggaag ggcgatcggt gcgggcctct tcgctataac gcccagctgg 60

[0089] cgaaaggggg atgtgctgca aaggcgatta aagttgggta acgccagggt tttcccagtc 120

[0090] acgacgttgt aaaacgacgg cccagtgagc gcgcgtaata cgactcacta taggcgaatt 180

[0091] gggtaccgggg ccccccctcg aggtcgacgg tatcgataag cttgatatcg aattcctgca 240

[0092] gcccggggga tcagatatac gacgtttttc cctttagcgc cggcccgctc aaattcttca 300

[0093] cgtgtatcag aaagcgcttt cgctatcggt gtccagccgg ttggtttaat cttgctgagt 360

[0094] gattgctcaa atggagtcac ggcgtacggc tgaaggccgt ataacgtttc tgtcgcgtta 420

[0095] catgatacgg ctttgcctga gtttttatta ttgcccttat gcccgaatac ccgaagcatc 480

[0096] agattcgtgt cttcaggaag aacttccgca aaagaagtga cagactcttt cgcaacatct 540

[0097] attttacgct ctccccctgt tttttgtatc atgctcccgc ttgcgtcaaa cagcacggcg 600

[0098] acgtttgttt cttcgtgctt tttctcagct gcaaaaacag gtgatccgaa gctggctcgt 660

[0099] aagcatca...

Embodiment 2

[0142] Embodiment 2: gene fragment cloning method of the present invention

[0143] Source of strain: Bacillus megaterium (Bacillus megaterium) Ap25 was screened in wheat rhizosphere soil, strain preservation number is: CGMCC No.1213.

[0144] The specific method is as follows:

[0145] 1. Detection of endoglucanase enzyme activity

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Abstract

The invention is a gene of huge bacillus endoglucanase and its preparing method. The invention uses SDS, proteinase K and CTAB / NaCl processes to extract DNA from huge bacillus genome, uses Sau3A to make partial enzyme cutting, connects gene fragment with linearized plasmid pBluescriptII, converts Escherichia coli DH5alpha, screens electropositive clonal seeds (74 strains) by endoglucanase enzyme activity detection, extracts the recombinant plasmids from the invert seeds with better enzyme activity expression, and obtains endoglucanase coded gene fragments amounting to 941bp. The gene fragment can dehydrate beta- bond at will along the amylase chain to release oligose, degrading the cell walls of pathogenic epiphyte, inhibiting the epiphyte growth and achieving the preventing and controlling effect.

Description

Technical field: [0001] The invention relates to a gene of Bacillus megaterium endoglucanase and a preparation method thereof, belonging to the field of biotechnology. Background technique: [0002] Dextranases are a class of hydrolytic enzymes that degrade dextran into glucose. Glucanases can be divided into β-1,2-, β-1,3-, β-1,4-, β-1,6-glucanases according to the different glycosidic bond sites. Glucanase can be divided into endoglucanase and exoglucanase according to the position of the cutting point. Exoglucanase mainly hydrolyzes the glucan residue broken by the non-reducing end. Glucanases can freely hydrolyze β-bonds along the polysaccharide chain to release oligosaccharides. The degradation of β-glucans in fungi is usually the result of the combined action of endo- and exo-glucanases. In fact, the presence of multiple glucanases is found in many microorganisms, and the hydrolysis of dextran is endoglucanase (EC3.2.1.39) and exoglucanase (EC3.2.1.58) Result of joi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/11C12N15/56C12N15/10
Inventor 杨合同黄玉杰李纪顺M·瑞德P·哈韦
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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