PCR技术基本原理及其在环境污染监测中的应用

聚合酶链反应简称ＰＣＲ，是美国ＰＥ－ｃｅｎｔｕｓ公司ＭｕＵｉｓ等人于１９８５年创立的，。ＰＣＲ技术创立以来，以特异性强，灵敏度高，操作简便，快速等优点，迅速渗透到生物学科的各个领域，大大提高了应用基因探针检测特异基因序称的能力。     

高效降解环己酮红球菌JDM-3-12的分离及其系统发育分析

从岳阳巴陵石化公司环己酮生产车间总出水口的污泥中筛选到一株环己酮降解菌株,菌号为JDM-3-12;该菌能以环己酮为唯一碳源且能忍受5000 mg/L的环己酮,当环己酮的质量浓度为2000 mg/L时,在温度为30℃,转速为150 r/m in,pH=7的条件下,72小时内该菌株对环己酮的降解率达到97.91%。通过形态观察、生理生化特征检测和基于16S rRNA基因序列的系统发育分析,初步鉴定其为赤红球菌(Rhodococcus ruber)中的一个菌株,该菌最适生长温度为35℃,最适生长pH 7。     

氯酚的生物降解特性及其微生物的16SrRNA基因序列分析

从受氯代有机物污染的土壤中富集分离到对2、4-二氯酚具有高效降解能力的微生物混合菌群。实验表明，降解1mol二氯酚可以定量释放出2mol的氯离子，在生物流化床反应器中，以聚胺酯泡沫块为固定化载体吸附固定化微生物，进行了连续降解氯酚的实验研究，当水力停留时间为24h，二氯酚的初始浓度为30μmol／L时，二氯酚的去除率均在90％以上，利用平板划线法从混合微生物菌群中分离到可以利用二氯酚为唯一碳源和能源的纯种微生物，16SrRNA基因序列分析结果表明，该微生物为Rhodococcus属。     

狗甲流病毒传染给人风险低

农业部前日发布消息，中国农业大学报告该校动物医学院临床医院从52份发病犬鼻咽拭子样品中，检出两份样品呈甲型H1N1流感病原学阳性。基因序列分析表明，该病毒与当前人群中流行的病毒同源性为99％。有关官员称，这是我国首例检测出狗感染甲流。     

Characterization of a salt-tolerant bacterium Bacillus sp. from a membrane bioreactor for saline wastewater treatment

High salt concentrations can cause plasmolysis and loss of activity of cells, but the salt-torlerant bacterium can endure the high salt concentrations in wastewater. In this research 7 salt-torlerant bacteria, which could survive in dry powder products and could degrade organic contaminants in saline wastewater, were isolated from a membrane bioreactor. The strain NY6 which showed the fastest growth rate, best property for organic matter degradation and could survive in dry powder more than 3 months was selected and characterized. It was classified as Bacillus aerius based on the analysis of the morphological and physiological properties as well as the 16 S rRNA sequence and Neigh borjoining tree. The strain NY6 could survive in the salinity up to 6% and the optimal growth salinity is2%; it belongs to a slightly halophilic bacterium. The capability of its dry powder products for COD removal was 800 mg COD/(g·day) in synthesized saline wastewater with salinity of 2%. According to salt-tolerant mechanism research, when the salinity was below 2%, the stain NY6 absorbed K+and Na+to maintain osmotic equilibrium, and when the salinity was above 2%, the NY6 kept its life by producing a large amount of spores.     

一种新厌氧氨氧化菌的16S rRNA基因序列测试

运用序批式试验测定厌氧氨氧化污泥的氨氮和亚硝态氮消耗量,求得厌氧氨氧化活性为9.84×10-4 mg·(mg·h)-1,厌氧氨氧化菌消耗NO-2-N与NJ 4-N之比为1.311;采用分子生物学方法从EGSB反应器颗粒污泥中提取细菌总DNA,经纯化、特异引物PCR扩增、克隆、测序等过程,得到厌氧氨氧化菌部分16S rDNA序列;通过系统发育树分析可以发现,在EGSB反应器中富集得到的厌氧氨氧化菌种(anaerobic ammonium-oxidizing Planctomycete cquenviron-1)与Candidatus"Anammoxoglobus propionicus"和Candidatus"Jettenia asiatica"同属,arnaerobic ammonium-oxidizing Planctomycete cquenviron-1与其他厌氧氨氧化菌基因序列的同源性最大为93%.结果表明,前期EGSB反应器富集得到了一种新型厌氧氨氧化菌,该菌株命名为anaerobic ammonium-oxidizing Planctomycete cquenviron-1.     

Identification and characterization of integron mediated antibiotic resistance in pentachlorophenol degrading bacterium isolated from the chemostat

A bacterial consortium was developed by continuous enrichment of microbial population isolated from sediment core of pulp and paper mill e uent in mineral salts medium (MSM) supplemented with pentachlorophenol (PCP) as sole source of carbon and energy in the chemostat. The consortia contained three bacterial strains. They were identified as Escherichia coli, Pseudomonas aeruginosa and Acinetobacter sp. by 16S rRNA gene sequence analysis. Acinetobacter sp. readily degraded PCP through the formation of tetrachloro-phydroquinone (TecH), 2-chloro-1,4-benzenediol and products of ortho ring cleavage detected by gas chromatograph/mass spectrometer (GC-MS). Out of the three acclimated PCP degrading bacterial strains only one strain, Acinetobacter sp. showed the presence of integron gene cassette as a marker of its stability and antibiotic resistance. The strain possessed a 4.17 kb amplicon with 22 ORF’s. The plasmid isolated from the Acinetobacter sp. was subjected to shotgun cloning through restriction digestion by BamHI, HindIII and SalI, ligated to pUC19 vector and transformed into E. coli XLBlue1 , and finally selected on MSM containing PCP as sole source of carbon and energy with ampicillin as antibiotic marker. DNA sequence analysis of recombinant clones indicated homology with integron gene cassette and multiple antibiotic resistance genes.     

润滑油降解菌的分离、鉴定及降解特性

以HVI 500润滑油为唯一碳源进行选择性富集培养,从油污染土壤中筛选出3株菌,分别命名为SN0901,SN0902和SN0903. 采用重铬酸钾法测定含HVI 500润滑油培养液的ρ(COD_Cr),用以评价分离菌对润滑油的降解能力. 结果表明,由于润滑油降解而使培养液ρ(COD_Cr)降低,即3株菌均为HVI 500润滑油降解菌. 根据形态学观察、生理生化试验和16S rDNA基因序列比对分析,初步确定3株菌分别属于假单胞菌属(Pseudomonas)、苍白杆菌属(Ochrobactrum)和博德特氏菌属(Bordetella). 采用倾注平板法对不同温度下降解菌的菌落计数,采用重铬酸钾法测定不同pH下培养液的ρ(COD_Cr). 结果显示,温度和pH对菌株降解作用影响显著,3株菌最适宜的降解温度为30～34 ℃,pH为6.0～7.8,但每株菌的最适宜降解温度和pH稍有不同.      

黄芪根瘤菌CA8561和JL84的部分16S rRNA基因序列的测定

报导了黄芪根瘤菌ＣＡ８５６１和ＪＬ８４的部分１６Ｓ ｒＲＮＡ基因序列，测算了它们与已知根瘤菌、茎瘤菌各属模式种之间的遗传距离。结果表明，ＣＡ８５６１和ＪＬ８４均具有独立的进化路线，充分显示了黄芪根瘤菌的遗传多样性。     

文摘

新冠病毒基因工程疫苗研发,已完成基因序列合成2月15日,中国医药集团有限公司在京披露,该集团与中国疾病预防控制中心合作,新型冠状病毒疫苗的研发已经取得进展。国药集团旗下的国药中生研究院启动了新型冠状病毒基因工程疫苗研发,现已完成了基因序列合成,正在进行重组质粒构建和工程菌筛选工作。据介绍,研究发现在新冠康复者血浆中,有着丰富的新型冠状病毒综合抗体。为此,国药中生在研究中还提出了“用新冠康复者血浆治疗危重患者”的建议,并已获得科技部紧急立项。目前国药中生已按照国家卫健委和国家药监局的标准制备了新冠康复者血浆,并用于11名危重病人治疗。