Pathogen_resistant Transgenic Plant of Brassica pekinensis by Transfering Antibacterial Peptide Gene and Its Genetic Stability

The soft rot infected by pathogenic bacterium Erwinia aroideae Holland is one of the three serious diseases of Chinese cabbage ( Brassica pekinensis Rupr.). By constructing vector system of high frequency transformation mediated by Agrobacterium tunefaciens EHA105, anti-bacterial peptide gene with strong bactericidal action to pathogenic bacteria was introduced into Chinese cabbage AB-81 self-bred line and the transgenic plants were obtained. PCR and Southern blotting detection showed that target gene was integrated into plant genome of Chinese cabbage. The tests of bacteriostasis action of the extract from transgenic plants in vitro, and the assay of disease-resistant of transgenic plantlets in the tube and the pot by perfusing inoculation with pathogenic bacteria showed obvious resistance to soft rot. This resistance can be a stable heredity by genetic analysis of generations of transgenic plants self-bred, separation ratio of its R, was 3:1. The resistance to Km and disease of soft rot was still kept in the R-5. These results indicated the possibility of breeding new varieties of anti-soft rot Chinese cabbage by transgenic plants as parents.

Isolation and Purification of Antibacterial Peptides from Mussel

[ Objective ] This study aimed to extract antibacterial peptides from mussel. [ Method ] Blue mussels were used as raw materials for direct extraction of antibacterial peptides by using O. 5 % acetic acid, and the antibacterial peptides were isolated and purified by Sephacryl S-100 polyacrylamide gel chromatography. The fractions were collected for measurement of antibacterial activity and minimum inhibitory concentration for a variety of bacterial species by filter paper diffusion assay. Molecular weight of the antibacterial peptides was determined by SDS-PAGE. Variation of antibacterial activity of antibacterial peptides was measured at 100 ~C under conditions of different processing time and different pH. [ Result] The O. 5% acetic acid was used for crude extraction of antibacterial peptides as extrac- tion solution and led to relatively high extraction efficiency. By using Sephacryl S-100, the antibacterial peptides could be purified as a single substance. The isola- ted and purified antibacterial peptides of mussel had relatively strong antibacterial properties with molecular weight of 5 908, showing heat-resistance acid-alkaline resistance. [ Conclusion] This study laid the theoretical foundation for large-scale production of antibacterial peotides.

Inducing and isolation of antibacterial peptides from oriental fruit fly, Bactrocera dorsalis Hendel

One antibacterial activity fraction from an immunized dipteran insect, Bactrocera dorsalis, was isolated and purified by prepurification, ion-exchange chromatography, gel filtration chromatography and reverse-phase high performance liquid chromatography （HPLC）. The final purified fraction was checked on the Smart system HPLC and was judged as a pure fraction. The results of physical and biological analysis revealed that this fraction is heat stable and showed strong activities against Gram-positive bacterial growth. It possesses antibicrobial peptide properties and is worth further investigation.

High yield synthesis of cyclic analogues of antibacterial peptides P-113 by Sortase A-mediated ligation and their conformation studies

P-113 is a fragment of natural occurring peptide Histatin 5 found in human saliva. This peptide exhibited broad spectrum of antibacterial and antifungal biological activities. In this study, bifunctional P-113 peptides 2–5 were designed as Sortase A substrates and synthesized by solid support peptide synthesis,where the N-terminus were equipped with glycine and its analogues, and C-terminus were extended with LPETGGS, respectively. Under Sortase A catalyzed condition, head to tail cyclization products 7–10were afforded in yields from 76% to 93%. The conformation insights of linear peptides 2–5 and cyclic analogues 7–10 in aqueous buffers and in trifluroethanol（TFE） analyzed by circular dichroism（CD）suggested that a-helix structures were produced progressively in hydrophobic environment independent of the cyclization, which displayed the similar behavior as parent peptide P-113.

Induction,selection and antibacterial activity of the antibacterial peptides from lepidopteran insect cultured cell lines

We induced 3 cell lines that were in vitro cultured from Lepidoptera with heat inactivated Escherichia coliDH_(5α) to stimulate the antibacterial peptide followed by antibacterial activity assay,induction dynamic research and Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis(Tricine SDS-PAGE)experiment.The antibacterial activity of the induced BTI-Tn-5B1 cell line was the highest,and the antibacterial activity increased gradually to the highest level in 16 hours after stimulation.A new antibacterial peptide with a molecular weight of about 8000 Da was preferentially induced in Trichoplusia ni BTI-Tn-5B1 cells in 16 hours after stimulation.Antibacterial activity assays indicated that it had inhibition against Staphylococcus aureus,Escherichia coli K_(12)D_(31) and Salmonella derby.It has especially strong inhibition against Gram-negative bacteria such as Escherichia coli K_(12)D_(31) and Salmonella derby.

Isolation and Preliminary Identification of Antibacterial Active Substances of Antagonistic Actinomycetes against Peach Crown Gall(Agrobacterium tumefaciens)

[ Objective ] The paper was to isolate and preliminarily identify the antibacterial active substances of antagonistic actinomyeete strain G19 obtained from the soil highly affected by peach crown gall （Agrobacterium tumefaciens）. [ Method] The antibacterial substances of antagonistic actinomycete strain G19 were ex- tracted using protein precipitation method, then isolated and purified using high performance liquid chromatography and medium-pressure preparative chromatogra- phy. Its molecular weight was determined by MALDI-TOFMS method, and the related functional groups were verified through chemical color reaction. [ Result] Seven peptide portions were produced from the antibacterial substances of antagonistic actinomycete strain G19 with the molecular weights of 900 - 1 300 Da after isolation and purification. It could be also inferred that it contained Cys, and carried with H2O and Na＋. Color reaction of functional groups verified that the sub- stance was polypeptide containing glycosyl. [ Conclusion] The result provided basis for the final definition of the structure of antibacterial substances in antagonistic actinomycete strain G19.

Application of antimicrobial peptides in production life

Antibacterial peptide is a small molecular weight,a substance existing in the biological body,plays a very important role in congenital immune systems.Today,in various antibiotics,the problem of drug resistance has caused people to live an indelible impact.Since the antibacterial peptide has spectral antibacterial properties,it has the characteristics of participating in human natural immunity and no residual,and is now one of the research and development hotspots.Antibacterial peptide is a recognized substance that can replace antibiotics,and people have never stopped,and new types of antimicrobial peptides have also emerged in the Volkswagen Vision.This paper is based on the use of antimicrosis peptides in daily life,briefly describes the effects of antimicrobial peptides in medical,animal husbandry and industrial.

Development of an antimicrobial peptide-loaded mineralized collagen bone scaffold for infective bone defect repair

The repair of infective bone defects is a great challenge in clinical work.It is of vital importance to develop a kind of bone scaffold with good osteogenic properties and long-term antibacterial activity for local anti-infection and bone regeneration.A porous mineralized collagen(MC)scaffold containing poly(D,L-lactide-co-glycolic acid)(PLGA)microspheres loaded with two antibacterial synthetic peptides,Pac-525 or KSL-W was developed and characterized via scanning electron microscopy(SEM),porosity measurement,swelling and mechanical tests.The results showed that the MC scaffold embedded with smooth and compact PLGA microspheres had a positive effect on cell growth and also had antibacterial properties.Through toxicity analysis,cell morphology and proliferation analysis and alkaline phosphatase evaluation,the antibacterial scaffolds showed excellent biocompatibility and osteogenic activity.The antibacterial property evaluated with Staphylococcus aureus and Escherichia coli suggested that the sustained release of Pac-525 or KSL-W from the scaffolds could inhibit the bacterial growth aforementioned in the long term.Our results suggest that the antimicrobial peptides-loaded MC bone scaffold has good antibacterial and osteogenic activities,thus providing a great promise for the treatment of infective bone defects.