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Cattle lactoferrin engineering bacterium and preparation method for antibiotic peptide cattle lactoferrin

A technology of bovine lactoferrin and bovine lactoferrin, which is applied in the biological field, can solve the problems of low expression level, inability to realize commercial production, and high cost, and achieve the effect of high expression amount

Inactive Publication Date: 2009-07-22
INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high cost of isolating bovine lactoferrin from natural milk, its commercial production cannot be realized. However, the fundamental restriction factor for the production of bovine lactoferrin by genetic engineering technology is that the expression product has a strong suicide effect on the host and the expression level is low.

Method used

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  • Cattle lactoferrin engineering bacterium and preparation method for antibiotic peptide cattle lactoferrin
  • Cattle lactoferrin engineering bacterium and preparation method for antibiotic peptide cattle lactoferrin
  • Cattle lactoferrin engineering bacterium and preparation method for antibiotic peptide cattle lactoferrin

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Experimental program
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Effect test

Embodiment Construction

[0039] Preparation of mutant photorhabdus luminescens TZR and construction of bovine lactoferrin expression plasmid pBAD-CipB-BLfcin-Ampin

[0040] 1 material

[0041] 1.1 Strain and carrier

[0042] See Table 1 for strains and vector genotypes and sources.

[0043] Table 1 Strain and carrier genotype and source

[0044]

[0045]

[0046] Note: AP r , ampicillin resistance; Apra r , Apramycin resistance; Genta r , Gentamycin resistance.

[0047] 1.2 Tool enzyme

[0048] RNaseA, Proteinase K, and restriction endonucleases were purchased from New England Biolabs. Taqpolymerase was purchased from Eppendorf Company.

[0049] 1.3 Kit

[0050] The PCR purification kit was purchased from Inviteck, and the small, medium and large extraction plasmid kits were products of QIAGEN. Bacterial Genomic DNA Extraction Kit is a product of TaKaRa Company. PCR2.1 TOPO TA cloning kit: purchased from Invitrogen (www.invitrogen.com). The Red / ET kit was provided by the German gene ...

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Abstract

The invention discloses a cow milk ferroprotein engineering bacterium CCTCC NO: M206132 and a preparing method of antibiotic peptide cow milk ferroprotein element, which comprises the following steps: resuscitating engineering bacterium photorhabdus luminescens TZR+ expressing plasmid pBAD-CipB-BLfcin-Ampin on LB flat; choosing monoclonal from LB medium; seeding medium photorhabdus luminescens TZR+ pBAD-CipB-BLfcin-Ampin in LB medium; inducing; centrifuging; collecting cell; hanging load the cell with solution I; transferring into new centrifuge tube; eccentric-collecting cell; cryolysis-splitting cell; property-separating CipB blend lactoferrin element; adding into parenzyme; enzymolyzing CipB blend cow milk lactoferrin element; delivering pure cow milk ferroprotein element; getting the product. This invention possesses highly effective expression and cheap cost.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically relates to a bovine lactoferrin engineering bacterium, and also relates to a method for preparing an antibacterial peptide bovine lactoferrin by the bovine lactoferrin engineering bacterium, and the antibacterial peptide is widely used in animal feed. Background technique [0002] When Horoaki et al. (1991) studied the thermal stability of apolactoferrin (lactoferrin, LfB), they found that LfB was obviously degraded under acidic conditions, but its antibacterial activity increased instead, so it was speculated that there is a resistance in Lf Thermal antibacterial active peptides are stable in acidic conditions, but unstable in neutral or alkaline environments. Tomita et al. (1991) studied the antibacterial effect of the protease hydrolyzate of Lf B. The results showed that the small molecule polypeptide obtained by hydrolyzing pig trypsin had a strong antibacterial effect, and its ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/09C12P21/02C12R1/01
Inventor 唐志如张友明印遇龙
Owner INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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