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@# Objective To analyze the epidemiological characteristics of mumps in Jiamusi city, and predict the incidence trend, so as to provide evidence for taking targeted prevention and control measures in the cold areas in Heilongjiang Province. Methods The mumps cases were collected from China Information System for Disease Control and Prevention in Jiamusi city from 2004 to 2017.Descriptive epidemiologic method was used for analyzing the epidemic of mumps in Jiamusi city during 2004-2017, and the autoregressive integrated moving average (ARIMA) was used to forecast the incidence of mumps in 2018. Results A total of 1 586 cases of mumps were reported in Jiamusi city during 2004-2017, the average annual incidence rate was 4.52/100 000. The incidence of mumps increased year by year from 2015 to 2017.The ratio of male to female was 1.67 :1,those aged from 3 to 19 years accounted for 86.32% of the total cases, the higher incidence rates were found at the age of 5-9 and 10-14 years.The incidence of mumps presented obviously seasonal characteristics.Most cases concentrated from April to July and from November to January. The incidence of urban disease was higher than that of other counties; The established finally mode was ARIMA(4,1,2)(2,0,0)12 and the predicted incidence from January 2018 to June 2018 was consistent with the actual one.From July 2018 to December 2018,predictive mumps incidence were:0.30/100 000, 0.35/100 000, 0.38/100 000, 0.39/100 000, 0.35/100 000, 0.33/100 000. Conclusions ARIMA model could predict the trend of mumps in Jiamusi city.To reduce the incidence of children, it is recommended to develop a second dose of mumps vaccine at the preschool age(3-6 years)or primary school;At the same time, surveillance and control should be continually strengthened in kindergartens and schools.
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AIM To establish an HPLC method for the simultaneous content determination of four constituents in Jinlan Xiaoyan Tablets (Lonicerae japonicae Flos,Paeoniae Radix Rubra,Salviae miltiorrhizae Radix et Rhizoma,etc.).METHODS The analysis of 80％ methanol extract of this drug was performed on a 30 ℃ thermostatic Wondasil C1s column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 327,230,286 and 250 nm.RESULTS Chlorogenic acid,paeoniflorin,salvianolic acid B and glycyrrhizic acid showed good linear relationships within the ranges of 46.48-464.84 μg/mL (r =0.999 5),20.28-202.75 μg/mL (r=0.999 1),12.44-124.35 μg/mL (r =0.999 6) and 15.09-150.89 μg/mL (r =0.999 9),whose average recoveries were 99.30％,99.10％,99.38％ and 100.27％ with the RSDs of 1.35％,1.37％,1.28％ and 0.86％,respectively.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Jinlan Xiaoyan Tablets.
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Epigallocatechin-3-gallate (EGCG) is not only the most active component of green tea, but the main object in green tea research. Numerous studies have shown that EGCG has cellular protec-tion functions, such as antioxidization, scavenging free radical , anti-cancer and antiapoptotic proper-ties. It has also been proved that EGCG could activate several signaling pathways. Nuclear factor erythroid-2p45 related factor 2/antioxidant response element (Nrf2/ARE) pathway is a key way to main-tain the redox homeostasis by regulating phase Ⅱ detoxification enzymes, and anti-inflammatory proteins. Recent studies have shown that EGCG plays an inportant role in the activation of Nrf2/ARE pathway in urinary, respiratory, cardiovascular, digestive systems. This review summarizes the research progress in EGCG as an activator of Nrf2/ARE signaling pathway, and provides reference for further research and exploitation of EGCG as potential medicine.
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AIM To investigate the effects of Shuxuening Injection (Ginkgo biloba leaf extract) on serum lactic acid (Lac),soluble CD14-st (Presepsin) and nitric oxide synthase (NOS) levels in sepsis patients.METHODS One hundred and eight patients with sepsis treated by routine treatment in our hospital from Jan.2014 to Oct.2016 were randomly divided into two groups,control group and Shuxuening group (therapy group).Two weeks were one therapeutic course.Before the treatment (the onset of patients within 3 hours),at 6 hours and 5 days after the treatment,Lac and Presepsin levels were detected,and the changes of nitric oxide (NO),NOS,inducible nitric oxide synthase (iNOS) and sequential organ failure assessment (SOFA) score were observed.Incidence of major adverse cardiac events (MACE) and 28-day survival were recorded at the same time.RESULTS Before the treatment,there were no significant differences in SOFA score and the levels of Lac,Presepsin,NO,NOS and iNOS between the two groups (P ＞ 0.05).Six hours after the treatment,the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05),both the two groups had lower levels of Lac and Presepsin than those before the treatment (P ＜ 0.05);five days after the treatment,the levels of Lac and Presepsin in the two groups were lower than those at 6 hours after the treatment (P ＜ 0.05),the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05).The SOFA score,NO,NOS and iNOS levels after the treatment in the therapy group were lower than those in the control group (P ＜ 0.05).The levels of Lac and Presepsin in sepsis patients were positively correlated with SOFA score (r =0.245,0.261,P =0.011,0.006).The patients in the therapy group had lower incidence of MACE and 28-day mortality rate than those in the control group (P ＜ 0.05).CONCLUSION The therapeutic effect of Shuxuening Injection combined with routine treatment on sepsis patients is superior to that of routine treatment,which can improve the prognosis of patients to a certain extent.
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AIM To investigate the effects of Shuxuening Injection (Ginkgo biloba leaf extract) on serum lactic acid (Lac),soluble CD14-st (Presepsin) and nitric oxide synthase (NOS) levels in sepsis patients.METHODS One hundred and eight patients with sepsis treated by routine treatment in our hospital from Jan.2014 to Oct.2016 were randomly divided into two groups,control group and Shuxuening group (therapy group).Two weeks were one therapeutic course.Before the treatment (the onset of patients within 3 hours),at 6 hours and 5 days after the treatment,Lac and Presepsin levels were detected,and the changes of nitric oxide (NO),NOS,inducible nitric oxide synthase (iNOS) and sequential organ failure assessment (SOFA) score were observed.Incidence of major adverse cardiac events (MACE) and 28-day survival were recorded at the same time.RESULTS Before the treatment,there were no significant differences in SOFA score and the levels of Lac,Presepsin,NO,NOS and iNOS between the two groups (P ＞ 0.05).Six hours after the treatment,the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05),both the two groups had lower levels of Lac and Presepsin than those before the treatment (P ＜ 0.05);five days after the treatment,the levels of Lac and Presepsin in the two groups were lower than those at 6 hours after the treatment (P ＜ 0.05),the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05).The SOFA score,NO,NOS and iNOS levels after the treatment in the therapy group were lower than those in the control group (P ＜ 0.05).The levels of Lac and Presepsin in sepsis patients were positively correlated with SOFA score (r =0.245,0.261,P =0.011,0.006).The patients in the therapy group had lower incidence of MACE and 28-day mortality rate than those in the control group (P ＜ 0.05).CONCLUSION The therapeutic effect of Shuxuening Injection combined with routine treatment on sepsis patients is superior to that of routine treatment,which can improve the prognosis of patients to a certain extent.
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Objective To explore the relationship between blood lipids and the severity of coronary artery disease. Methods A total of 4 751 patients with chest pain were divided into two groups according to the results of coronary angiography: coronary artery disease group (n = 3 402 stenosis in one main branch stenosis ≥ 50%) and control group (stenosis<50%, n = 1 349). According tothe number of coronary artery stenosis, the coronary artery disease group was divided into three subgroups (group 1: 1branch stenosis; group 2: 2 branch stenosis; and group 3: 3 or more than 3 branch stenosis). The serum total cholesterol (TC), triglycefides (TG),high density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were determined and LDL-C/HDL-C was calculated. The relationship between lipids and the severity of coronary artery stenosis was analyzed by Spearman correlation analysis, and the risk factors of coronary artery disease were analyzed by multiple-stepwise logistic regression analysis. Results The levels of TG, TC, LDL-C and the ratio of LDL-C/HDL-C in the coronary artery disease group were significantly lower than those in the control group (P<0. 01). LDL- C/HDL-C ratio increased with the increase of diseased branches in the disease group (P<0. 001), and the number of diseased branches was positively correlated with the ratio of LDL-C/HDL-C (R = 0. 25, P<0. 01). Logistic multiple regression analysis showed that high LDL-C/HDL-C level was an independent risk factor for coronary heart disease. Conclusion LDL-C/HDL-C ratio is an independent risk factor for coronary heart disease and it is positively correlated with the severity of coronary artery lesions.
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Microbiology experiment existing independently from microbiology theoretical curriculum is an indispensable compulsory course in contemporary life science. This article presents the principle applied by the National Excellent Microbiology Course teaching group in Nankai University, which is to strengthen the undergraduates’ basic skills of conducting microbiology experiments. With an aim to enhance the core competitiveness of the undergraduates, we have established the three-level experimental contents. A new multilevel teaching pattern focusing on basic skill training as the cornerstone has been applied to enhance the overall competences of the students and to stimulate their innovation abilities. Students’ experimental accomplishment will also be taken into consideration when their experiment results are evaluated, which helps to standardizing their research ethics.
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A strain Mortierella isabellina M6-22-4, which was sensitive to hygromycin B, was selected by treating parental spores with N-methyl-N' -Nitro-N-nitrosoguanidine (MNNG). Protoplasts of the strain Mortierella isabellina M6-22-4 were transformed successfully to hygromycin B resistance using the PD4 plasmid, which contains the Escherichia coli hph gene under the control of Mortierella alpina his H4.1 promoter. The PD4 plasmid was introduced by PEG/CaCl2 treatment. Transformation frequencies of 1.6 - 2.8 transformants/microg of DNA were achieved. Then they were successively incubated to non-selected PDA plates for 10 generations. About 31.6% transformants only from digested plasmid were mitotically stable and showed different hygromycin B resistance when they were incubated back to selection plates. The results of PCR and Southern analysis in three transformants indicated that the plasmid PD4 had been integrated into the fungal genome with 1 - 2 copies. This is the first report of Mortierella isabellina transformation system and supplies an important tool for further research into genetic manipulation of this filamentous fungus.
Subject(s)
Anti-Bacterial Agents , Pharmacology , Blotting, Southern , Drug Resistance, Microbial , Genetics , Escherichia coli Proteins , Genetics , Genome, Fungal , Genetics , Hygromycin B , Pharmacology , Mortierella , Genetics , Plasmids , Genetics , Polymerase Chain Reaction , Protoplasts , Metabolism , Transformation, GeneticABSTRACT
Pseudomonas aeruginosa is an important opportunistic human pathogen. It encodes many virulence factors and one of them is type III secretion system (TTSS). Effectors proteins can be delivered into host cells directly by this system, causing necrosis or apoptosis. popN gene is the first gene in the popN operon of TTSS gene cluster. To investigate its function, popN gene deletion mutant was generated in this study, and we found this mutant can secrete effectors proteins constitutively under non-inducting condition in DMEM medium containing serum. The results indicated that PopN is a negative regulator of the TTSS expression. However, no secreted effector proteins were detectable when the popN- mutant was grown in LB medium under non-inducting condition. To investigate the possible reasons, effects of growth status and protease (s) inhibitors on the TTSS were investigated. We present evidences that indicate protease mediated degradation of secreted effector proteins played a key role in the phenotypic inconsistency of popN- mutant.
Subject(s)
ADP Ribose Transferases , Metabolism , Bodily Secretions , Bacterial Proteins , Genetics , Metabolism , Bodily Secretions , Bacterial Toxins , Metabolism , Gene Expression Regulation, Bacterial , Mutation , Peptide Hydrolases , Genetics , Metabolism , Pore Forming Cytotoxic Proteins , Genetics , Bodily Secretions , Protease Inhibitors , Pharmacology , Pseudomonas aeruginosa , Genetics , Metabolism , VirulenceABSTRACT
Thamnidium elegans is a kind of phycomycete that produces essential unsaturated fatty acids, particularly y-linolenic acid. In this process, delta6-Fatty acid desaturase (D6D) plays a key role due to its enzymatic properties that catalyze the delta6 site dehydrogenation of precursor linoleic acid (18:2delta(9, 12) n-6) and a-linolenic acid (18:3delta(9, 12, 15) n-3). This reaction is the first and rate-limiting step of highly unsaturated fatty acids (HUFA) synthesis pathways. After we have isolated and cloned the gene coding delta6-fatty acid desaturase from Thamnidium elegans As3.2806 (GenBank accession number DQ099380), our interest focuses on the promotion and regulation of the gene transcription. To achieve this aim, we designed long primers and used nested inverse PCR to amplify DNA flanking sequences. First, genome of Thamnidium elegans was extracted and digested with restriction enzymes EcoR I and Kpn I , respectively. Then we ligated the digested DNA with T4 ligase at low concentration which is propitious for linear DNA to joint intromolecule. According to the sequence of delta6-fatty acid desaturase gene of Thamnidium elegans, we designed a couple of 35nt long inverse primers and two couples of shorter inverse primers for inverse PCR. Three rounds of PCR reactions were performed. In the primary reaction, the ligated DNA was used as a template, and the product was used as the template of the secondary reaction, the tertiary reaction was achieved in the same way. After all the three rounds of reactions, we got a nice product about 4 kb from the EcoR I digested sample, in which a 1.3kb 5' upstream sequence (GenBank accession number DQ309425) of delta6-fatty acid desaturase gene containing several putative regulatory elements including TATA. box, FSE-2, AP-1 sites, CCAAT cis-element site and STRE-binding site was derived after sequencing. All of these implied intensely that this 1.3kb fragment is a condition-regulated promoter. It is the first report about Thamnidium elegans detla6-fatty acid desaturase gene promoter. The procedure described here is a rapid and simple method and particularly useful to isolate flanking sequences from fungal genome. box, FSE-2, AP-1 sites, CCAAT cis-element site and STRE-binding site was derived after sequencing. All of these implied intensely that this 1.3 kb fragment is a condition-regulated promoter. It is the first report about Thamnidium elegans delta6-fatty acid desaturase gene promoter. The procedure described here is a rapid and simple method and particularly useful to isolate flanking sequences from fungal genome.
Subject(s)
Base Sequence , Cloning, Molecular , DNA Primers , Linoleoyl-CoA Desaturase , Genetics , Molecular Sequence Data , Mucorales , Genetics , Polymerase Chain Reaction , MethodsABSTRACT
Delta6-fatty acid desaturase is a membrane-bound enzyme, which is rate-limiting for the biosynthesis of polyunsaturated fatty acids. A cDNA sequence putatively encoding a delta6-fatty acid desaturase was isolated from Rhizopus arrhizus NK300037 using RT-PCR and RACE methods in our previous work. Sequence and function analysis indicated that this sequence was a novel delta6-fatty acid desaturase gene which had an open reading frame of 1377bp coding 458 amino acids of 52kD. The methylotrophic yeast Pichia pastoris, has been developed into a highly successful system for the production of a variety of heterologous proteins during the past 20 years. In this work, the Rhizopus arrhizus delta6-fatty acid desaturase gene (RAD6) was subcloned into expression vector pPIC3.5K to generate a recombinant plasmid pPICRAD6, which was subsequently transformed into Pichia pastoris strain GS115 for heterologous expression by electroporation method. Total fatty acids were extracted from the induced cells and methylated. The resultant fatty acid methyl esters (FAME) were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Fatty acids analysis showed that the coding product introduced a new double bond at delta6 position of appropriate fatty acid substrates including C16:1, C17:1, C18:1, linoleic acid and alpha-linolenic acid without chain length specificity of fatty acids. Furthermore, modification of sequence flanking AUG codon of this delta6-fatty acid desaturase gene increased the expression of target gene in P. pastoris. All of these results suggest that P. pastoris is an optimal expression system of delta6-fatty acid desaturase gene.
Subject(s)
Cloning, Molecular , Electroporation , Fungal Proteins , Genetics , Genetic Vectors , Linoleic Acid , Metabolism , Linoleoyl-CoA Desaturase , Genetics , Pichia , Genetics , Metabolism , Rhizopus , GeneticsABSTRACT
Gamma-linolenic acid (GLA, C18:3delta6 ,9,12), an essential polyunsaturated fatty acid, plays an important role in hormone regulation and fatty acid metabolization. Delta6-fatty acid desaturase (D6D) is the rate-limiting enzyme of the desaturation of linoleic acid (C18:2delta9,12) in the production of gamma-linolenic acid. A deficiency of GLA may have occurred when delta6-fatty acid desaturase activity decreases in aging, stress, diabetes, eczema, and some infections. To establish a new expression system for delta6-fatty acid desaturase gene in Pichia pastoris, which is an increasingly popular heterologous gene expression system, a gene encoding delta6-fatty acid desaturase from Mortieralla alpina was isolated by PCR amplification. The PCR product was then digested by EcoR I and Not I and subcloned into the intracellular expression vector pPIC3.5K to generate the recombinant vector pPIC3.5K-MA6. The resulting vector was linearized by Sac I and electroporated into P. pastoris SMD1168 (his- pep-) host cells. After electroporation, aliquots were spreaded on the MDS plates and incubated at 30 degrees C for three days until colonies appeared. Those transformants were subsequently screened for clones with high copy number by using the YPD plates containing G418. To identify the D6D constructs that were produced, chromosomal DNA of the transformants were prepared and used as template for PCR with the primer 5' AOX and 3' AOX. The PCR product of Mut+ recombinants was shown as a band of 1.38 kb of D6D gene and the product of 2.2 kb of AOX1 gene, while the product of Mut(s) transformants only was shown as a band of 1.38 kb of the D6D gene.To further confirm the transformants containing a functional D6D gene, the positive clones were selected and induced by methanol for expression. Those induced cultures were taken for analyses of the intracellular fatty acid composition by GC. The resultant chromatograms of fatty acid methyl esters showed that a novel peak was detected, which was not apparent in the case of control. Comparisons of the retention times of the newly yielded peaks with those of authentic standards have anticipated that the fatty acid is GLA. And this prospects was positively supported by definitive assignments of the compounds by GCMS analyses. Thus, the active delta6-fatty acid desaturase was expressed intracellularly in P. pastoris and gamma-linolenic acid reached 16.26% of the total fatty acid in recombinant P. pastoris strains. It was the first report about the expression of Mortieralla alpina D6D gene in P. pastoris.
Subject(s)
Cloning, Molecular , Fatty Acids , Gas Chromatography-Mass Spectrometry , Linoleoyl-CoA Desaturase , Genetics , Mortierella , Genetics , Pichia , Genetics , Plasmids , gamma-Linolenic AcidABSTRACT
Polyunsaturated fatty acids (PUFAs) including gamma-linolenic acid are valuable products because of their involvement in several aspects of human health care. GLA has been claimed to play a crucial role in development and prevention of some skin diseases, diabetes, reproductive disorder and others. At present, market demand for most gamma-linolenic acid is growing continually and current sources are inadequate for satisfying this demand due to the significant problems of low productivity, complex and expensive downstream process and unstable quality. Therefore, seeking for alternative sources are demanding. delta6-fatty acid desaturase is the rate-limiting enzyme for the biosynthesis of PUFAs, which catalyses the conversion of linoleic acid and alpha-linolenic acid to gamma-linolenic acid and stearidonic acid respectively. Unfortunately, the structure information on membrane desaturases is scarce because of the technical limitations in obtaining quantities of purified protein and the intrinsic difficulties in obtaining crystals from membrane proteins. With the isolation of the genes coding for delta6-fatty acid desaturase from various organisms, its characteristics will be elucidated gradually. Here we concisely reviewed the recent progress on studies of molecular biology including the cloning of delta6-fatty acid desaturase gene, structure and function, phylogeny and prospects of gene engineering application.
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Cloning, Molecular , Fatty Acid Desaturases , Genetics , Metabolism , Fatty Acids, Unsaturated , Genetic Engineering , Methods , Phylogeny , gamma-Linolenic AcidABSTRACT
Gamma-linolenic acid (GLA, C18:3delta6.9.12) is nutritional and important polyunsaturated fatty acid in human and animal diets. GLA play an important role in hormone regulation and fatty acid metabolization. Furthermore it is also the biological precursor of a group of molecules, including prostaglandins, leukotrienes and thromboxanes. Vast majority of oilseed crops do not produce GLA, but linoleic acid (LA, C18:2delta9.12) as its substrate. GLA is only produced by a small number of oilseed plants such as evening promrose ( Oenotheera spp.), borage (Borago officinalis) and etc. delta6-fatty acid desaturase (D6D) is the rate-limiting enzyme in the production of GLA. It can convert from linoleic acid to linolenic acid. To produce GLA in tobacco, plant expression vector was first constructed. To facilitate preparation of plant expression constructs, flanking Xba I and Bgl II restriction enzyme sites were added to the coding region of clone pTMICL6 by PCR amplification. pTMICL6 contains delta6-fatty acid desaturase gene cloned from Mortierella isabellina which is an oil-producing fugus. The PCR product was purified and subcloned into the plant expression vector pGA643 to generate the recombinant vector pGAMICL6 which contains the ORF of the D6D gene of Mortierella isabellina, together with regulatory elements consisting of the cauliflower mosaic virus 35S promoter and the nopaline synthase (nos) termination sequence. The plasmid pGAMICL6 was transformed into Agrobacterium tumefaciens strain LBA4404 by method of freeze thawing of liquid nitrogen. Transformants were selected by plating on YEB medium plates containing kanamycin and streptomycin and grown overnight at 28 degrees C, then transformants were further identified by PCR. The positive transformant containing the plant expression vector pGAMICL6 was transformed into tobacco ( Nicotiana tabacum cv. Xanthi) via Agrobacterium infection. Transgenic plants were selected on 100 microg/mL kanamycin. Plants were maintained in axionic culture under controlled conditions. Total nucleic acids were extracted and purified from anti-kanamycin transgenic tobacco and were analysed by PCR. 48 out of 80 transgenic plants were positive, in other words, transformation efficiency is 60% . This shows that Mortierella isabellina D6D gene is transformed into tobacco. Genomic DNA from PCR positive transgenic tobacco plants was digested with Hind III restriction enzyme and fractionated by agarose gel electrophoresis. Southern blotting was performed with strandard procedures for vacuum transfer of nucleic acids to nylon membrane. The probe was delta6-fatty acid desaturase gene from M. isabellina, which was labeled with DIG-dUTP via random-primed labeling. Hybridization and immumological detection were carried out the kit of DIG detection. The result shows single hybridizing bands in each of the transgenic tobacco plants DNA, but no hybridization was observed to non-transgenic tobacco. This indicates that delta6-fatty acid desaturase gene is integrated into the genome of transgenic tobacco. To provide further evidence that the introduction of the M. isabellina cDNA into the tobacco genome was responsible for the novel desaturation products, total RNA was isolated from GLA-positive transgenic tobacco plants via both PCR and Southern blotting and separated by electrophoresis through 1% formaldehyde agarose gel. Northern blotting including probe labeling, hybridization and detection was the same as Southern blotting in operation approach. A positive hybridization signal of identical mobility was obtained from RNA isolated from the transgenic tobacco plants, but not from the control tobacco plant. At last, total fatty acids extracted from the positive transgenic tobacco were analyzed by gas chromatography (GC) of methyl esters to confirm the transgenic tobacco containing a functional delta6-fatty acid desaturase gene. The result shows that two peaks were observed in the chromatogram of FAMes. GLA and octadecatetraenoic acid (OTA, C18:4delta6.9.12.15) respectively have 19.7% and 3.5% of the total fatty acids in the transgenic plant. The presence of both GLA and OTA indicates that the delta6-fatty acid desaturase used both linoleic acid and a-linolenic acid (ALA, C18:3delta6.9.12.15) as substrates, and this may be responsible for the decrease in ALA observed in the transgenic line. That was the first report about the expression of M. isabellina delta6-fatty acid desaturase gene in tobacco. All results mentioned above have laid the foundation of the thorough studying on an breeding transgenic oilseeds containing GLA to change the fatty acid composition of conventional oilseeds, it is significant to study on regulation mechanism of fatty acid desaturase.
Subject(s)
Agrobacterium tumefaciens , Genetics , Fatty Acid Desaturases , Genetics , Metabolism , Genetic Vectors , Genetics , Mortierella , Genetics , Plants, Genetically Modified , Genetics , Metabolism , Polymerase Chain Reaction , Nicotiana , Genetics , Metabolism , gamma-Linolenic AcidABSTRACT
Genomic DNA of two fungi Thamnidium elegans and Umbelopsis isabellina were extracted with an amended Cetyltrimethyl Ammonium Bromide (CTAB) method. This modified method uses repeated freezing in liquid nitrogen and thawing with combination of shocking with glass beads to replace of the tra- ditional method. Quality and concentration of DNA extracted by the modified methodwere tested. Compared with the traditional method, higher yield and purity of genomic DNA were obtained with less amount ofmy- celium. The result indicted that this is a simple and highly efficient method, which is suitable to treat many samples at one time and for basic molecular experiments, such as restriction endonuclease reaction and PCR.
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Microbiology is an important,fundamental and obligatory course in contemporary life science.This article introduces that teaching group of microbiology in Nankai University realizes transformation of teaching center,fully embodies the modernization of teaching notion and gives full play to students' main effect practically by adhering to teaching reform as center,optimizing teaching method as measure,communicating in and after class and using multi-media and teaching web.Therefore,teaching system is established to adapt to modern teaching notion and eventually microbiology course becomes a cultivation platform to foster elites with both solid fundamental theory and innovating mind.