溶藻弧菌asp基因在大肠杆菌中表达活性研究及条件优化   总被引：1，自引：0，他引：1  

蔡双虎  鲁义善  吴灶和  简纪常  汤菊芬  黄郁葱 《广东海洋大学学报》2008,28(3)

为进一步研究溶藻弧菌碱性丝氨酸蛋白酶(ASP)蛋白生物学活性和功能,将构建的pET23d-asp在大肠杆菌BL21(DE3)中表达,对表达产物进行纯化分析,并优化表达条件。结果表明:在咪唑洗脱缓冲液浓度为100 mmol/L时,表达的可溶性ASP被大量洗脱,纯化的ASP相对分子质量约为42 000,具有蛋白活性;在诱导温度28℃、异丙基-β-D硫代半乳糖苷(Isopropyl-β-D-thiogalactopyranoside,IPTG)浓度1 mmol/L及诱导时间16 h时,表达的活性ASP蛋白最多。  相似文献   

养殖近江牡蛎致病弧茵的分离与鉴定     

张占会  张其中  李春勇  陈浩如  张吕平 《热带海洋学报》2008,27(6)

从广东省疫区患病近江牡蛎Crassostrea ariakensis消化盲囊及血液中分离纯化出10株细菌，人工注射感染实验发现其中3株细菌注射组死亡率显著高于对照组。3株细菌均为革兰氏阴性，短杆状，能运动。常规生理生化和Biolog细菌鉴定系统测试结果表明，菌株A3G-2与溶藻弧菌Vibrio alginolyticus亲缘关系最近，菌株A2G-5B与解蛋白弧菌V．proteolyticus亲缘关系最近，菌株B1X-1与鲨鱼弧菌V．carchariae亲缘关系最近。为了进一步确定它们的分类学地位，分别测定了3株弧菌的16S rDNA序列，分析相关细菌相应序列的同源性，并构建系统进化树。结果表明，菌株A3G-2与溶藻弧菌亲缘关系最近，菌株A2G-5B与解蛋白弧菌亲缘关系最近，菌株B1X-1与鲨鱼弧菌亲缘关系最近。因此菌株A3G-2、A2G-5B和BIX-1分别被鉴定为溶藻弧菌、解蛋白弧菌和鲨鱼弧菌。组织病理观察发现患病牡蛎主要受损器官是消化盲囊，表现为腺泡管内壁吸收细胞崩解脱落，管腔扩大，进而管壁解体，消化盲囊出现大面积受损。  相似文献   

红鳍笛鲷弧菌病病原的研究   总被引：1，自引：0，他引：1  

《广东海洋大学学报》2004,24(6):1-5

  相似文献   

A culture-dependent survey of thermophilic bacteria from hot springs in Xiamen area in China     

YANG Bo  OUYANG Jianping  AO Jingqun  CHEN Xinhua 《海洋学报(英文版)》2009,28(1):62-71

Microbes are believed to play important roles in ecosystem function in many environments. The hot springs of Xiamen Island are close to the Xiamen Sea, and may have some characteristics different from those of inland hot springs. Microbes living in the hot springs of Xiamen may have new characteristics. However, little is known about microbial communities of hot springs close to the Xiamen Sea. A culturedependent survey of microbial population in the Xiamen hot springs was performed by using an approach combining total cellular protein profile identification and 16S rRNA gene sequencing. A total of 328 isolates of bacteria were obtained from liquid and sediment samples from the Xiamen hot springs, including neutrophilic thermophilic bacteria and moderately thermophilic acidophiles. Neutrophilic thermophilic bacteria, which grow at a temperature range of 55—90℃ including Rhodothermus marinus (Strain 1), Thermus thermophilus (Strain 2), Thermus thiopara (Strain 3), Geobacillus stearothermophilus (Strain 4), Geobacillus thermoleovorans (Strain 5), and Pseudomonas pseudoalcaligenes (Strain 6), were recovered by 2216E plates. Moderately thermophilic acidophiles, which can grow at temperatures above 50℃ and a pH range of 1.8—3.5 such as Alicyclobacillus acidoterrestris (Strain 8), Sulfobacillus acidophilus (Strain 9), and Sulfobacillus thermosulfidooxidans (Strain 10), were isolated on selective solid medium containing sulfur and Fe2+. Among these strains, Rhodothermus marinus, Thermus thermophilus and Geobacillus stearothermophilus are not only thermophiles, but also halophiles. One bacterium strain (Strain 6) shared 99% nucleotide sequence homology with Pseudomonas pseudoalcaligenes on the 16S rRNA gene sequence, but was quite different from Pseudomonas pseudoalcaligenes in biological characteristics, suggesting that it may represent a novel thermophilic species. Results indicated that various species of neutrophilic thermophiles and moderately thermophilic acidophiles were widely distributed in the Xiamen hot springs and that Rhodothermus marinus and Thermus thermophilus dominated the cultivable microbial community.  相似文献   

Growth of marine bacteria and ammonium regeneration from substrates in different C: N ratios     

WANG Jiangtao  YIN Xiaonan 《海洋学报(英文版)》2009,28(3):59-64

Natural assemblages of marine bacteria were chosen in a batch culture experiments. The impact of varying nitrogen substrate concentrations and the substrate C:N ratios (C:N_S) on the bacterial C:N ratio (C:N_B), the bacterial growth efficiency (BGE) and ammonium regeneration was mainly examined. The C:N_S ratios varied from 5:1 (carbon limitation) to 40:1 (nitrogen limitation) with varying combinations of glucose and NO₃^-. The C:N_B ratio had positive relationship with the C:N_S ratio (r=0.93, n=8), whose value was 3.77 when the C:N_S ratio was 5:1 but increased to 6.47 when the C:NS ratio was 40:1. These results indicate that the C:N_B ratio is a potential diagnostic tool for determining the bacterial growth in natural waters controlled by either, carbon or nitrogen. BGE decreased with the declining nitrate concentration and negatively related to C:N_s (r=-0.51, n=8). The average value of BGE was 0.20. This value was a little lower than other reports, which could be induced by the nitrogen source used in our experiments. Finally, regeneration time of ammonium delayed with the increasing C:N_S ratio, which indicates that there were different metabolism mechanisms when bacterial growth was limited by carbon source and nitrogen source.  相似文献   

Isolation and identification of bacteria associated with the surfaces of several algal species     

王子峰  肖天  逄少军  刘敏  岳海东 《中国海洋湖沼学报》2009,27(3):487-492

We conducted this study to assess the diversity of bacteria associated with the surfaces of algae based on 16S rDNA sequence analyses.Twelve strains of bacteria were obtained from the surfaces of the following four species of algae:Gracilaria textorii,Ulva pertusa,Laminaria japonica,and Polysiphonia urceolata.The isolated strains of bacteria can be divided into two groups:Halomonas and Vibrio,in physiology,biochemical characteristics and 16S rDNA sequence analyses.The phylogenetic tree constructed based on ...  相似文献   

拮抗香蕉枯萎病菌海洋细菌的筛选和鉴定   总被引：3，自引：0，他引：3  

黄瑶  杨耿周  张雅娟  何秋韵  林巧玲  易润华 《广东海洋大学学报》2009,29(6):72-77

从93份中国南部海域近海海泥样品中分离出1800多株海洋细菌,以对峙培养法,测定海洋细菌对香蕉枯萎菌的拮抗作用,再以琼脂划线培养法(agar streak)测定菌株的抑菌活性,结果表明:约55.6%的菌株对香蕉枯萎菌有一定的拮抗作用,其中58株菌株抑制香蕉枯萎菌生长所形成的抑菌带宽度大于10.0mm,抑菌作用较好,复筛后其中13个菌株形成的抑菌带宽在15.0mm以上。经反复转接和对峙培养,筛选出拮抗效果好、抑菌活性稳定的菌株TC-1,初步鉴定为芽孢杆菌属的细菌,TC-1对14种病原微生物都有较好的抑菌效果。  相似文献   

2009年冬、夏季南海北部超微型浮游生物的分布特征及其环境相关分析   总被引：2，自引：1，他引：1  

乐凤凤  蔡昱明  孙军  郝锵  刘诚刚  翟红昌  陈建芳 《海洋学报》2015,37(12):41-55

2009年2月(冬季)和8月(夏季)在南海北部海域(nSCS)采用流式细胞术对聚球藻、原绿球藻、超微型光合真核生物3类超微型光合浮游生物和异养浮游细菌的丰度和碳生物量的时空分布特征进行了研究,并分析了其与环境因子之间的关系。结果表明,夏季聚球藻和原绿球藻的平均丰度高于冬季,超微型光合真核生物和异养浮游细菌的丰度反之,为冬季高于夏季。聚球藻、超微型光合真核生物和异养浮游细菌在富营养的近岸陆架海域丰度较高,而原绿球藻高丰度则出现在陆坡开阔海域。在垂直分布上,聚球藻主要分布在跃层以上,跃层以下丰度迅速降低;原绿球藻高丰度主要出现在真光层底部;超微型光合真核生物在水层中的高值同样出现在真光层底部,且与Pico级份叶绿素a浓度分布一致;异养浮游细菌在水体中的分布与聚球藻类似。这些分布格局的差异,取决于环境条件的变化和4类超微型浮游生物生态生理适应性的差异。在超微型光合浮游生物群落中,各类群碳生物量的贡献因季节和海域类型的不同而发生变化:聚球藻在夏季近岸陆架区占超微型光合浮游生物总碳生物量的41%,原绿球藻在陆坡开阔海成为主要贡献者(50%),超微型光合真核生物碳生物量以冬季为高(在近岸陆架区占比68%)。冬、夏季异养浮游细菌碳生物量均高于超微型光合浮游生物碳生物量。  相似文献   

北冰洋楚科奇海浮游细菌丰度和生产力及其分布特征     

高源  何剑锋  陈敏  林凌  张芳 《海洋学报》2015,37(8):96-104

2012年夏季中国第5次北极科学考察期间,对北冰洋楚科奇海及其北部边缘海浮游细菌丰度和生产力进行了测定,并将其与环境因子进行了相关性分析。结果显示,楚科奇海浮游细菌丰度的变化范围为0.56×108~6.41×108 cells/dm3,平均为2.25×108 cells/dm3;细菌生产力介于0.042~1.92mg/(m3·d)(以碳计)之间,平均为0.54mg/(m3·d)(以碳计),与已有研究结果基本相当。陆架区细菌丰度和生产力要明显高于北部边缘区,但前者的单位细菌生产力则较低。与环境因子的相关性分析显示,细菌丰度与温度和叶绿素a浓度存在显著正相关(p0.01),表明北极变暖导致的海水升温及浮游植物生物量的增加均会促进细菌的生长,从而进一步提高细菌在海洋生态系统和碳循环中的作用。但陆架区的细菌生产力与环境参数均没有显著相关性,表明其影响因素较为复杂;生产力在北部边缘区则仅与叶绿素a存在显著正相关(p0.01),表明浮游植物生长过程产生的溶解有机碳(DOC)是细菌生长最为主要的碳源,碳源的单一可能制约细菌的生产从而导致该海域无冰状态下细菌丰度的增加不如预期,但融冰过程带来的大量DOC将促进细菌活性的增加。  相似文献   

青岛潮间带沉积物可培养厌氧细菌多样性的研究   总被引：1，自引：1，他引：0  

王雅苹  史晓翀  于少兰  张晓华 《海洋科学》2015,39(3):92-99

为了验证设计的简易厌氧培养方法,作者以青岛潮间带沉积物为研究对象,采用5种培养基,从青岛潮间带沉积物共分离获得138株厌氧细菌。16S r DNA序列分析表明,这些细菌分属3个门15个属32个种,其中γ-变形菌纲64株18个种,在种类上处于优势地位;此外还包括δ-变形菌纲(δ-Proteobacteria)16株2个种,ε-变形菌纲4株1个种,拟杆菌门(Bacteroidetes)29株8个种,梭杆菌门(Fusobacteria)25株3个种。在属水平上,弧菌属(Vibrio)、Marinifilum属、泥杆菌属(Ilyobacter)、脱硫弧菌属(Desulfovibrio)、希瓦氏菌属(Shewanella)在数量上占优势。此外,有26株8种细菌(占总菌株数的18.84%)与已知菌的同源性介于89.38%~94.22%,为潜在的海洋细菌新科或新属,表明此简易厌氧菌培养方法在获得新菌方面具有较大优势。另外,研究结果还表明,不同培养基对特定的类群具有选择性:2216E培养基对γ-变形菌纲分离培养效率较高;SPG培养基在获得新菌方面具有优势(占新菌数62.5%),且这些新菌大多属于拟杆菌门和梭杆菌门,其中SPG-1培养基对于分离硫酸盐还原菌和难培养的ε-变形菌纲细菌具有优势,SPG-4培养基对分离硝酸盐还原菌具有优势。  相似文献