Abstract:A high Streptolydigin-producing strain Streptomyces lydicus AS 4.2501-L8 with genetic stability was obtained from the original strain S. lydicus AS 4.2501-P28 by mutagenesis with UV and acridine orange, followed by resistance selection. Its yield of Streptolydigin was 177.0 μg/mL, 96.2% higher than the original strain.

Abstract:A mathematical model, describing the population growth under limited conditions, was developed based on the time dependent changes of the specific growth rate. This model could simulate the lag growth phase, the exponential growth phase and the stationary growth phase of the population growth of many kinds of biology or their organs or cells. This model had less parameters, and all the parameters had clear physiological meanings and were easy to be calculated.

Abstract:Eighty-eight strains of endophytic fungus，belonging to 28 genera, were isolated from wild Polygala tenuifolia Willd of two different growth stage，and the inhibitory activities screening to three species of microbe were conducted research. The results showed that the quantity, population and distribution of the endophytic fungi varied in different tissue and different growth stage of P. tenuifolia. The species of endophytic fungus distributed in stem were more than in root and leaf, and the dominant genus was Alternaria Nees. Seventy-three strains, accounting for 83.0% of all the isolates, presented antimicrobial activities to the tested indicators. High antimicrobial activities were found in four strains, which seperatedly belonged to the genera as follows: Trichothecium Link,Cephalosp-orium Corda,Alternaria Nees,Trichosporiella Kamyschko. et al.

Abstract:The epitope-G₁ gene, cloned from the pMD-G plasmid including G protein gene of bovine ephemeral fever virus, was subcloned into expression vector pPIC9K to construct pPIC9K-G₁ recombinant plasmid successfully, and the recombinant plasmid linearized was transformed into Pichia pastoris GS115 by electroporation. The recombinant Pichia pastoris strains were screened by G418 and PCR, and induced by methanol. The expressed products were analyzed by SDS-PAGE, deglycosylation, Western blot, ELISA, immunizing rabbits and specificity experiments. The results indicated that the gene was expressed successfully in GS115 and glycosylated moderately, and the target protein had nicer biological activity and specificity. The protein is able to be used as coating antigen to develop ELISA Kit for diagnosing bovine ephemeral fever.

Abstract:In order to isolate genes related with the osmoadaptation and glycerol metabolism of Candida glycerinogenes, a transformation system based on the dominant selectable marker Zeocin and restriction enzyme-mediated integration (REMI) was established. Effects of seven restriction enzymes on transformation efficiency of C.glycerinogenes were tested. Transformation conditions were optimized in the presence of Hind III. Under the optimal conditions of OD₆₀₀≈1.3, voltage of 1.5 kV, 2.0×10⁹ competent cells/mL, 100 units of Hind III added, the transformation efficiency was up to 129 trnaformants/μg DNA. 58% of transformants were stable on nonselective medium. These results suggest that REMI technique would be beneficial to the genetic transformation of C.glycerinogenes.

Abstract:It was found that the phosphate concentration effected significantly on the cell growth and the production of aminoglycoside antibiotic JI-20A. Although initial phosphate concentration of 6.1mmol/L～9.6 mmol/L was beneficial for the cell growth, the production of aminoglycoside antibiotic JI-20A was inhibited. When the phosphate concentration was controlled below 1.1 mmol/L in the biosynthesis phase of aminoglycoside antibiotic JI-20A, high alkaline phosphatase activity and low pyruvic acid concentration were resulted in, and the production of aminoglycoside antibiotic JI-20A was enhanced.

Abstract:This paper investigated the effects of carbon sources on the growth of Chlorella zofingiensis and the production of astaxanthin. The result showed that the concentration of 20g/L was beneficial to the growth of Chlorella zofingiensis, but on the this concentration biomass and astaxanthin content was low; on the concentration of 50g/L, the growth is slower, but biomass and astaxanthin content was high . Among the three carbon sources investigated , sucrose and glucose were more beneficial to growth and production of astaxanthin than fructose. When 50g/L sucrose was used , the astaxanthin content and astaxanthin yield was 0.94 mg/g and 9.61 mg/L respectively.

Abstract:On the basis of morphological, physiological and biochemical characters as well as sequence analysis of 16S rDNA, the antagonistic bacterium strain 3-1-16 was identified. Under optics microscope, the strain was bacilliform, G⁺, stained evenly purple. Furthermore, many granulose structures were found in cell without parasporal crystal under EM observation. According to Biolog Bacteria Identified system, 3-1-16 strain has 98% similarity with Bacillus megaterium MO31. A 1443-bp 16S rRNA gene sequence was obtained from strain 3-1-16. The BLAST search of the GenBank database using this sequence showed its similarity to many species of Bacillus. It was 99.4% similar to the 16S rRNA gene sequence of Bacillus megaterium strain MO31 (GenBank accession No. AY553118.1) over 1437 bases. Phylogenetic tree of Bacillus 16S rRNA gene sequences showed that 3-1-16 strain was clusterred with three Bacillus megaterium. Based on those characteristics, strain 3-1-16 was identified as Bacillus megaterium GIMA 1.001. Glasshouse trials demonstrated that the control efficiency of 3-1-16 strain against tomato bacterial wilt disease was 81.3%

Abstract:Carotenoids play an important role in regulating the hydrogen production of hydrogen-producing Rhodobacter sp. The carotenoids of hydrogen-producing Rhodobacter sp. grown in acetate medium were extracted by using acetone-methanol (7∶2，V/V) solvent and were separated by using thin-layer chromatography on silica-gel plate. The qualitative and quantitative of the carotenoids were analyzed by spectrometry. The results showed that the carotenoids were completely extracted three times with acetone-methanol (7∶2，V/V) in two hours. The ultrasonication had little effect on yield of carotenoids. The yield of carotenoids was 2.81mg/g wet cell. There were 4 spots on the silica-gel plate in the order of yellow, red, light red and light yellow. Yellow spot and red spot were the dominant composition of carotenoid in Rhodobacter sp. The spectrometry data showed that the yellow and red component might be the spheroidene and spirilloanthin respectively.

Abstract:The present investigation was undertaken in order to select the surface-sterilization technique most efficient for eliminating epiphytes, to document endophytes of healthy tissues from Glaycyrrhiza inflat Bat. in Xinjiang. Surface sterilization with 5% commercial solution of sodium hypochlorite for 5 minutes was reaffirmed as adequate for removing epiphytes on licorice roots. From the 151 segments incubated, 149 bacterial isolates and 2 fungal isolates were obtained. From all the isolates, Bacterial isolates were identified by VITEK-AMS. Part of Bacteria were identified in 13 different genus. Fungal species were characterized as Penicillium sp. and Fusarium sp.with microscope.

Abstract:A fusion protein of glucagons-like peptide-1 and human serum albumin (GLP-1/HSA) was expressed and secreted into the fermentation broth with recombinant Pichia pastoris. The productivity of expressed GLP-1/HSA could reach 63.6mg/L in 10L fermentor. After concentrated with hollow-fiber ultrafiltration membrane, GLP-1/HSA was purified from fermentation broth by hydrophobic chromatography, negative ion exchange chromatography and gel filtration chromatography in turn. The HPLC analysis showed that the purified GLP-1/HSA had an overall purity of 95.8%. Furthermore, the analysis of in vivo activity indicated that GLP-1/HSA had the bioactivity of native GLP-1, and could significantly reduce blood glucose level 4h after intraperitoneal administration. It was concluded that a great deal of GLP-1/HSA with higher purity could be harvested by Pichia pastoris expression system and the established purification methods. Preliminary studies show a new potential for developing the long-acting GLP-1 analogs for clinical applications.

Abstract:The extracellular metabolism products of the marine bacterium CⅢ-1, a endophytic marine bacterium isolated from mangrove (V.C Bruguiera gymnorrhiza), were extracted with dichloromethane and then analyzed by GC/MS technique. The results showed that the mainly chemical composition and their concentrations of the extracellular metabolism of this strain were arachic acid（10.12%）, all-trans-squalene（7.33%），12-tricosanone（7.07%），2-(2-hydroxyethoxy)ethyl stearate（6.21%），tetramethyl-oxirane（4.67%）, pyrrole-2-carboxylicacid-4-(1-chlorodec-1-enyl)-3,5-dimethyl-ethylester(4.63%）, 3-methyl-3-undecene（3.55%） and so on. Among them the all-trans-squalene has an important value of application.

Abstract:In order to reveal the differences among the dimorphic cells, basidiospores, arthrospores and mycelia of Tremella fuciformis were subjected to microscopical observation and electrophoresis analysis. Morphology and nucleus phase showed that arthrospores are larger than basidiospores in diameter, basidiospores are mononuclear cells, the majority of arthrospores are dikaryocytes and the mononuclear cells are in minority, mycelia are dikaryocytes with typical clamp connection. The whole protein and isozyme analysis indicated that the mycelial form contains more bands than that of yeast form and some differences in peroxidase and esterase between the two forms were found, while the polyphenol oxidase are almost similar. Therefore, corresponding changes in metabolism level are occurred during the form transition of the dimorphic cell in T. fuciformis.

Abstract:The dynamic of endophytic bacteria at different growth stage of tomato and use of these endophytic bacteria to control tomato bacterial wilt were studied. The results showed that endophytic bacteria could be found in the tomato seeds and their quantities reached the highest peak in the adult plants both in resistant and susceptible cultivars. The amount of endophytic bacteria in adult plants of resistant tomato cultivars was 2.43×10⁵CFU/g FW in the root and 22.9×10⁴ CFU/g FW in the stem, while the amount of endophytic bacteria in adult plants of susceptible tomato cultivars was 9.8×10⁴CFU/g FW in the root and 13.4×10⁴CFU/g FW in the stem respectively. Seventeen strains of endophytic bacteria from resistant cultivars and only seven strains from susceptible cultivars were found to be antagonistic to Ralstonia solanacearum. In addition, some strains of endophytic bacteria had the abilities of promoting tomato seed germination and controlling tomato bacterial wilt, among which, strain 5R and 3R had better control effect of 91.7% and 81.3% respectively.

Abstract:NP gene of AIV was amplified by PCR and cloned into the expressing plasmid pSOC, where NP gene was fused to 3′end of T4 phage small outer capsid gene encoding SOC protein. T4 phage expression vector named pSOC-NP was constructed and used to transform E.coli BL21(DE3). The recombinant E.coli BL21(DE3) was induced by IPTG. The expressed fusion protein SOC-NP was detected by SDS-PAGE with expected molecular size of 58 kD and the expression product accounted for 20.34% of the total bacterial protein. The immunological test applying Western blot indicated that SOC-NP fusion protein could react to AIV specifically.

Abstract:By using PDA plate and PDB shaking culture method, the effect of six chemical pesticides on the spore germination, plate inhibition and Biomass of the biological control strains Pochonia chlamydospora ZK7 for tobacco root-knot nematodes were measured. Results showed that strain ZK7 is resistant to six pesticides at lower concentration and the resistance level are different among different pesticides. Carbetamide and thiophos of them have the biggest resistance, next is phoxim-methyl and metaldehyde, the last is carbophenothion and aldoxycarb.

Abstract:A duplex real-time PCR, based on Taqman hybridization probes technology, were developed and applied to detect enterohemorrhagic Escherichia coli O157.Two pairs of primer and two probes were designed for detection rfbE and stx2 genes .the rfbE probe was 5′end labeled with FAM) and 3′end labeled with Taqman-MGB. The stx2 probe was 5′end labeled with VIC and 3′end labeled with Taqman-MGB. The detection limits of the sensitivity assays were 101 copies/uL of DNA; the qualitative consensus PCR assay indicated all Escherichia coli O157 were found rfbE positive and did not detect DNA from non- O157 isolates. In the duplicated experiment, coefficients of variation intra-assay and inter-assay over the dynamic range of the MGB probe assays were lower than 70% and 80%, respectively. These results show that this duplex real-time PCR can detect the Escherichia coli O157 and stx2 gene from samples rapidly, and that the virulence of the strains were known.

Abstract:D-D4FC (β-D-2′，3′-didehydro-2′，3′-dideoxy-5-fluorocytidine)，a new anti-HIV drug，is on its PhaseⅡ clinical trials in America，France and Germany. Our lab has synthesized D-D4FC successfully using N-deoxyribosyltransferase from Lactobacillus helveticto catalyzing the ribose transfer from D4T (β-D-2′，3′-unsaturated thymidine) to 5-FC (5-fluorocytidine).The yield of D-D4FC reached 25%.We discovered the reaction could also be done by using intact cells.The yield could increase to 50% in 12.5 hours and more convenient to industrial continuous process.In this paper，the conditions including pH，buffer，substrates concentration，cells amount，reaction time and a possible catalytic mechanism were studied and discussed.

Abstract:Induction and antagonistic activity of chitinases produced by Gliocladium catenulatum HL-1-1 were studied. Results showed that the isolate could produce chitinases under various culture conditions. Sclerotia powder could induce the isolate to produce much more chitinases. The optimum conditions to produce the enzyme were pH 4.5 at initial medium and to culture the isolate for 6 days. The chitinases were proved to inhibit the conidial germination of Fusarium nivale, Coniella diplodiella, Curvularia lunata and Alternaria alternata and the sclerotial germination of Sclerotinia sclerotiorum and Rhizoctonia solani obviously.

Abstract:The potential of Rhizobium sp. N613 to produce the exopolysaccharide (REPS) was studied in this paper. Using an orthogonal design in a flask-shaker culture system, the fermentation medium and conditions of synthesizing REPS were optimized. Based on these results, the fermentation kinetic parameters were obtained in the batch fermentation with a 10L fermentor. The REPS yield of 11.31g/L was achieved by metabolic regulation during 40 h fed-batch fermentation. Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect. The result of anti-tumor activities showed that inhibition rate was 53.40%, when dose of REPS was 5mg/kg. These results indicate that REPS has the following properties: the short duration of fermentation, the high yield, the low cost, the effective immunocompetence and thickening. Thus, REPS has the value of development and application.

Abstract:Phospholipase D (PLD) may play an important role in the infection and inflammation process, but it is unknown whether cellular PLD was activated during the invasion of Listeria monocytogenes into Vero cells. Our study demonstrated that firstly the Vero cells either infected by Listeria monocytogenes or stimulated by PMA shows remarkable increase on PLD activity, compared to basal ones (no stimuli). Further, in the Vero cells with the overexpression of catalytically inactive PLD₂ (mPLD₂-K758R), PLD activition by both Listeria monocytogenes and PMA were dramatically inhibited, leaving basal PLD activity unaffected. These results indicated that the PLD activities may increase while Listeria monocytogenes invade the Vero cells and PLD₂ is likely to be activated. The important physiological role of PLD in this pathogen_host interaction process need to be further studied.

Abstract:The effect of dark tea fermentation liquid with Eurotium Cristatum on the activity of digestive enzyme was researched, as fellowing amylase, protease, lipase. The results showed that dark tea fermentation liquid with Eurotium Cristatum may increase remarkably the activity of α-amylase and protease, but decrease efficiently the activity of lipase. The fermentation liquid improves the digestion and absorption of starch and protein, but inhibits the decomposition and absorption of fat, so it can explain the mechanism of Fu-brick tea's health functions.

Abstract:By streak plate method, a strain of heterotrophic microorganism which was extremely acidophilic, alkalitolerant, named by HJM, was isolated from an acid mine drainage of a certain copper mineral in Jiangxi Province. This strain was able to grow under pH value of 1.5～10.0. Morphological characteristics and sequences analysis of 16S rDNA and 26S rDNA D1/D2 region revealed that it belonged to the species P.guilliermondii. Resistant experiment to metals showed that it could resist copper of 45mmol/L, so it was an important strain used to study the resistance mechanism of copper.

Abstract:In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.

Abstract:Based on the strain breeding theory and metabolic engineering theory, A high-yield mutant of Lactobacillus Thermophilus ATCC8317 was obtained through the compound inducements by the original Acetic acid-Sodium acetate plate and the productivity increased 210%.The best media components included saccharifying corn,malt powder 30g/L,peptone 5g/L.Based on the variety of specific cell growth rate and specific L-lactic acid production rate at different temperatures, the strategy of temperature control was obtained. The total product of L-lactic acid reached 135g/L besides the rate of glucose consumed and the average L-lactic acid productivity were up to 95% and 2.25g/(L·h) respectively.

Abstract:Recombinant plasmids pHY-PA, pBBR-PA were constructed in which genes pdc and adhB were placed under the control of tac promoter, respectively, and had successfully expressed in Escherichia coli. Then these recombinant plasmids were electroporated into Lactobacillus strains for ethanol production. Preliminary ethanol fermentation using these Lactobacillus strains and their recombinants was carried out using 42℃ as fermentation temperature. The results indicated that introducing pdc and adhB, ethanologenic pathway was successfully constructed in L.plantanum CICIM B0080. 0.4% (V/V) ethanol was detected at the end of fermentation with 6.7% glucose, and that is 67-fold as the wild-type B0080. Two-fold of ethanol production was detected in L.amylovorus B0112 (pHY-PA) and L.acidophilus B0068 (pBBR-PA). Introducing both pdc and adhB, and meanwhile knock-outing the lactate dehydrogrnase gene may better convert carbon flux to ethanologenic direction.

Abstract:Using 16S rDNA clone library method, the bacteria composition of the Organic Matter-decomposing Inoculants A and B were investigated. The results indicated that: Sample A was clustered into 14 taxonomic operational units (OTUs),the dominant communities are Weissella confuse,Bacillus subtilis and Bacillus pumilus, which accounted for 28.6%,30.4% and 23.2%;Sample B was clustered into 43 OTUs,the dominant communities are Lactobacillus buchneri, Lactobacillus farciminis and Lactobacillus acetotolerans, which accounted for 18.03%,18.86% and 13.12%, respectively. The results had much difference from the samples' labels:there were not bacteria in the lable of sample A and only Bacillus pumilus was the same with the lable of sample B.This study showed that 16S rDNA clone library method has nicer application perspectives in analying the bacteria of microbial inoculant and its quality control.

Abstract:Class Ⅱa bacteriocins can be considered as the major subgroup of bacteriocins from lactic acid bacteria, not only because of their large number, but also because of their activities and potential applications. They have first attracted particular attention as listericidal compounds and are now believed to be the next in line if more bacteriocins are to be approved in the future. The present review attempts to provide an insight into general knowledge available for class Ⅱa bacteriocins and discuss common features and recent findings concerning these substances.

Abstract:Integrins are a family of cell surface glycoproteins that contribute to a variety of biological functions, including cell growth, migration, proliferation and morphology. In addition, integrins also play the important roles in pathological process. Several viruses have been showed to use integrins as receptors or co-receptors to infect host cells.This article mainly reviews the progress on integrins and their roles in FMDV infection.

Abstract:γ-Hexachlorocyclohexane (γ-HCH) is an organochlorine insecticide,it has been listed recalcitrant worldwide pollutant due to its toxicity, environmental persistence. Microorganisms capable of degrading γ-HCH have been isolated, several genes encoding the enzymes involved in the degradation of γ-HCH have been cloned and characterized. In this article, we summarized the current progress regarding γ-HCH biodegradation, with emphasis on the diversity of microorganisms degrading γ-HCH, the biodegradative pathway and relevant genes and enzymes.

Abstract:The detections of foodborne bacterial pathogens are the key technological link for the foodborne disease prevention and control. The PCR technology has been introduced for the pathogenic bacteria detections recent years and the many target genes have been found including the genes of various virulence, engymes and some specific species. These genes are frequently used in the PCR detection and the application of diagnostic kits.

Abstract:During invasion of Aspergillus fumigatus into host cells，there is an obvious cellular actin cytoskeleton rearrangement. Toll-like receptors (TLRs)，one of the most important PRRs （pattern recognizing receptors）, can tigger the intracellular signaling transduction to induce the innate immunity, meanwhile be involved in regulating bacteria-associated cellular actin cytoskeleton rearrangement. The PAMPs(pathogen-assosiated molecular patterns) of Aspergillus fumigatus can be recognized by TLR2 and TLR4, nevertheless, the role of TLRs in the regulation of cellular actin cytoskeleton rearrangement during invasion remains unclear. Further studies on the internalization of Aspergillus fumigatus through TLRs will contribute significantly to our understanding of regulation on invasion of Aspergillus fumigatus, eventually provide some ways to cure the fungal infection.

Abstract:Nitrobenzene is one of the toxic compounds. Much work had focused on biodegradation of it sofar. Two main pathways for nitrobenzene biodegradation, oxidative and partial reductive pathways, were reviewed in this article. The mechanism of these pathways including involved enzymes and genes was introduce in details. Comparative analysis of the pathways would provide basis for the development and application of biodegradation technology for nitrobenzene and other organic pollutants.

Abstract:The process and mechanism of chitin degradation in Vibrios were introduced in this paper by reviewing of recent progress in chitin catalytic cascade research. There exist three steps in chitin degradation, hydrolysis of chitin, transportation of chitin oligosaccharides and N-acetylglucosamine, and further degradation of chitobiose and N-acetylglucosamine-6-phosphate. Most of the reactions in chitin degradation are regulated by two-component signal transduction system.

Abstract:Avian influenza viruses (AIV) can cause serious economic losses and threaten to human health. The laboratory methods for rapid and accurate detecting AIV ensure timely implementation of intervention strategies so that it plays a key role in avian influenza prevention and control. The laboratory technologies for detecting AIV are topic subject and developed quickly those days. This paper reviews the advances of laboratory diagnosis technologies for AIV at 3 aspects of virus isolation, immunoassay and molecular diagnostics.

Abstract:Some types of bacteria swim through rotating their flagella. The swimming mechanism of bacteria during flagella bundling and tumble process is analyzed. The effects of body rotation and flagellum′s polymorphic transitions on bundling processes and the wall effect phenomenon are also discussed. Finally, based on dynamics similarity, a new microrobot module is put forward to further studying the flagella swimming phenomena. The research would be very helpful for constructing the bionic swimming robots under the low Reynolds number.

Abstract:Marine actinobacteria are novel sources for drug discovery and active natural products. Since the functional diversity of active metabolites from marine actinobacteria originates from the Bio-diversity of this important group of gram-positive bacteria, studies on the diversity of culturable marine actinobacteria are of great importance. In this review, the progress in the diversity studies of marine actinobacteria, especially the research in the marine sponge-associated actinomycetes, deep sea actinomycetes and the indigenous marine actinomycetes was reported. Meahwhile, the isolation techniques including the pre-treatment of samples and the selection of media were highlighted. The cultivation of as yet unculturable marine actinobacteria was discussed, and the importance of establishing regional centers for marine actinobacteria strain and gene resources was emphasized.

Abstract:By analysis of the “man-oriented” viewpoint, the paper revealed that teach students in accordance of their aptitude and advancing in proper sequence were the specific expressions and implementations in higher education. Expounding the concrete methods and measures from “teaching” and “learning” two aspects combined with Microbiology teaching practice, that classroom teaching is basically students centered, the teacher should carry out the forms of school running, course contents based on the needs of students and their ability to learn, and it is important for college students study that teacher must play the guiding role in learning, because of classroom learning must be combined with experimental learning practice and knowledge should be advanced by steps and making steady progress, which further shows that comprehensively carry out the education principle of the “man-oriented” is special importance or significance to microbiology teaching in higher education.

Abstract:The experimental teaching method and content of fermentative project in the past can not adapt to the development of biotechnology industry, at the same time can not meet the needs of training talented person of biotechnology.Therefore, studying from the experimental course content of fermentative project and taking reforming by experimental methods, to deep the student′s cognition and understanding of experimental course knowledge in entire fermentative project. Ultimately, it can foster the student′s experimental ability of independent character, comprehensiveness and innovation, reach the purpose of training the person with applying and innovative talented.

Abstract:The study compared the traditional medical microbiology experimental teaching with a new experimental teaching pattern,with the students majoring in anaesthesia as the research object. The new pattern mainly deals with the cultivation of the students' creativity by reforming and exploring the plan,the content,the method of experimental teaching and the ways of checking the students' work, adding general and designing experiment,and working out the PPT of the experiments.The result shows the new experimental teaching pattern contributes to the cultivation of the students'abilities of performing experiment, the ways of thinking, creativity and comprehensive analysis. It's better than the traditional experimental teaching pattern.A new medical microbiology experimental teaching systerm which is suit to the students majoring in anaesthesia has been established.

Abstract:To reform the teaching purpose and to incite the students' interests in medical bacteriological experiments，the comprehensive, designing and creative experiments with science research and exploring study were set up to culture the students to acquire performing, science thinking and expressing ability, and to master the basic technical ability and basic principle as well as acquainting with the main method and means of the medical microbiology research. The author tried an attempt in training the creative talents with creative ability and thinking.

Abstract:A direct lysis manner for extracting DNA directly from rhizosphere soil of transgenic cotton SGK321 and Zhongmiansuo 41 was designed by glass bead-lysozyme-SDS method. The results showed that intact DNA fragments over 20 kb were gained from rhizosphere soil planted two cultivars of transgenic cotton. DNA extracted suited to PCR amplification and RFLP.

Abstract:The crude toxin was extracted from hypha and culture solution of Phyllosticta commelimecola through three different polarity solvent: benzinum, puncificatum ethyl acetate and chloroform. The result indicated that the toxin secreted by Phyllosticta commelimecola not only was in hypha but also in culture solution and the extracting effect of ethyl acetate was the best. The soybean median and PSK media can be respectively used as solid and liquid culture media to produce toxin and grow mycelium. The optimal cultural conditions for producing toxin were temperature 32℃,cultured period 14d, cultured ways shaking of 150r/min.

Abstract:A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.