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1.
The culture of Sertoli cells has become an indispensable resource in studying spermatogenesis. A new Sertoli cell line (POSC) that consisted predominantly of fibroblast-like cells was derived from the testis of the olive flounder Paralichthys olivaceus and sub-cultured for 48 passages. Analysis of the mtDNA COI gene partial sequence confirmed that the cell line was from P. olivaceus. Cells were optimally maintained at 25°C in DMEM/F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, and epidermal growth factor. The growth curve of POSC showed a typical “S” shape. Chromosome analysis revealed that the cell line possessed the normal P. olivaceus diploid karyotype of 2n=48t. POSC expressed dmrt1 but not vasa, which was detected using RT-PCR and sequencing. Immunocytochemistry revealed that the cells exhibited the testicular Sertoli cell marker FasL. Therefore, POSC appeared to consist of testicular Sertoli cells. Bright fluorescent signals were observed after the cells were transfected with pEGFP-N3 plasmid, with the transfection efficiency reaching 10%. This research not only offers an ideal model for further gene expression and regulation studies on P. olivaceus, but also serves as valuable material in studying fish spermatogenesis, Sertoli cell-germ cell interactions, and the mechanism of growth and development of testis.  相似文献   
2.
We cloned and sequenced a prtV-like gene from Vibrio anguillarum M3 strain. This prtV gene encodes a putative protein of 918 amino acids, and is highly homologous to the V. cholerae prtV gene. We found that a prtV insertion mutant strain displayed lower gelatinase activity on gelatin agar, lower protease activity against azocasein, and lower activity for four glycosidases. This prtV mutant strain also had increased activity for two esterases in its extracellular products, as analyzed by the API ZYM system. In addition, the prtV mutant strain exhibited decreased growth in turbot intestinal mucus and reduced hemolytic activity on turbot erythrocytes. Infection experiments showed that the LD50 of the prtV mutant strain increased by at least 1 log compared to the wild-type in turbot fish. We propose that prtV plays an important role in the pathogenesis of V. anguillarum.  相似文献   
3.
采用体外测定细菌浓度、胞外产物(ECP)蛋白含量和蛋白酶活力的方法,进行了鳗弧菌M3菌株在2216E培养基中的培养条件研究。结果表明,该菌株用固体培养基培养至24h左右,可得到较高的菌体浓度、ECP蛋白含量和蛋白酶活力。采用响应面分析方法设计实验,用SAS统计软件分析数据,得到NaCl浓度、pH值和温度对菌体生长及蛋白酶产量影响的回归模型。在2216E培养基的基础上,添加不同氮源、碳源物质以及不同浓度蛋白胨进行生长研究。结果表明,胰大豆蛋白胨能促进菌体生长及ECP蛋白分泌;NH4Cl与酪蛋白水解物可抑制蛋白酶的产生;牙鲆肌肉匀浆对菌体、ECP蛋白产量和蛋白酶产生有不同程度的促进作用;培养基中蛋白胨浓度为4%时菌体量与ECP蛋白含量达最高值,在蛋白胨浓度为2%时蛋白酶分泌量已稳定;1%的葡萄糖、蔗糖、甘油均能显著地提高菌体及ECP蛋白产量,却抑制了蛋白酶的产生。  相似文献   
4.
间接ELISA技术在病原性鳗弧菌SMP1快速检测中的应用   总被引:2,自引:0,他引:2  
以大菱鲆(Scophthalmus maximus)出血症的病原菌——鳗弧菌(Vibrio anguillarum)SMP1为抗原,制备兔抗血清,测定了抗血清的工作浓度和对抗原的特异性,建立了在大菱鲆肝肾组织中检测SMP1的间接ELISA方法。SMP1人工感染大菱鲆,用建立的间接ELISA对大菱鲆肾脏和肝脏进行检测,结果肾脏的阳性检出率为87.7%,肝脏的阳性检出率为90%。该方法的建立有助于快速准确地鉴定由SMP1引起的大菱鲆疾病。  相似文献   
5.
After the single-strain abilities of organic-pollutant-degrading bacteria in bioremediation of sediment environment of shrimp culture are determined, the multistrain degrading effect of the compositions of different strains is measured. The results indicate that the multi-strains groups have higher degrading ability than the single-strain groups. Three-strain groups are better than two-strain groups, and fourstrain groups are better than three-strain groups and five-strain groups, the groups composed of strains Lt7222, Lt7511. Fc6308 and Gy7018 has the best degrading effect, the CODMn removal rate is 73.2 % in 66 h, and gets to 82.7% in 114 h. 30% higher than that of the best single-strain group; Groups of Lt7222. Lt7511. Lt7451 and Gy7018 are the second, whose CODMn removal rate is 82.1% in 114 h. It is suggested that multi-species bacteria be used as thnctional bacteria in bioremediation of mariculture environment.  相似文献   
6.
为了实现所筛选的对虾养殖环境生物修复作用菌的产业化生产,本文选择三株具有高效降解有机污染物能力的菌株为代表,对作用菌的发酵生产和菌体收集工艺进行了优化。结果表明,作用菌的最适发酵培养基配方(g/L)为蛋白胨 25,酵母粉 5,磷酸铁 0.2,灭菌前培养基的 pH 可先调为 8.0,搅拌方式为搅拌1h 间隔 1h, 出罐时间以 20h 为宜。菌体收集以等电点沉降法效率最高、成本最低,其中菌株 Lt7222 的最适沉降 pH 为 3.67、Gy7018 为 4.02、Lt7511 为 3.40。菌体回收率可达 90 %,回收后菌体的存活能力几乎不受影响。  相似文献   
7.
We cloned and sequenced a prtV-like gene from Vibrio anguillarum M3 strain.This prtV gene encodes a putative protein of 918 amino acids,and is highly homologous to the V.cholerae prtV gene.We found that a prtV insertion mutant strain displayed lower gelatinase activity on gelatin agar,lower protease activity against azocasein,and lower activity for four glycosidases.This prtV mutant strain also had increased activity for two esterases in its extracellular products,as analyzed by the API ZYM system.In additi...  相似文献   
8.
对虾养殖池环境修复作用菌固定化的研究   总被引:10,自引:0,他引:10  
为了将对虾养殖池环境修复作用菌Lt 7511投放到虾池底部降解有机污染物,并且不造成二次污染。以沸石作为固定化载体,研究了影响固定化的因素、固定化菌降解有机物的性质及保藏方法。结果表明,沸石在500℃灼烧2h可除去绝大部分吸附水分,增大表面积;固定化过程中沸石粒度、菌液的浓度及体积对沸石吸附作用菌的能力都有影响,每kg 60目沸石可吸附0.46g菌体(干质量);沸石固定化菌对饵料浸出液的降解效果明显高于非固定化菌;真空保藏的固定化菌的存活情况明显优于保藏在空气中的固定化菌。  相似文献   
9.
17β-羟类固醇脱氢酶1(17β-HSD1)的主要作用是将雌酮(El)转化为发挥功能的雌二醇(E2)。作者从牙鲆(Paralichthys olivaceus)性腺转录组数据库获得该基因的开放阅读框(ORF)序列,对其进行了验证,并分析了该基因在高温、外源性激素处理条件下性腺分化期性腺组织中的差异表达以及c AMP和转录因子(NR5a2和NR0b1)在精巢原代细胞中对该基因表达的作用。结果显示,牙鲆17β-HSD1基因的ORF为873bp,编码290个氨基酸,与其他鱼类的有很高的相似性。半定量RT-PCR结果表明,该基因在卵巢中高表达,精巢有少量表达,并且在雌性个体的鳃、头肾、肾、脾、胃和肠中也有表达。实时定量RT-PCR结果显示,该基因在卵巢或精巢分化的关键时期表达量较高;在精巢原代培养细胞中,外源信号分子c AMP及转录因子NR5a2可以显著下调17β-HSD1基因的表达(P0.05),且呈现剂量效应,转录因子NR0b1对该基因的调控也与剂量有关。作者推测牙鲆17β-HSD1基因在性腺分化中起一定的作用,其表达受到调控因子的作用,这些结果将有助于增加对鱼类性腺分化和发育的认识。  相似文献   
10.
本文通过qPCR方法,研究了我国重要海水鱼类——牙鲆(Paralichthys olivaceus)雌、雄成体组织以及性腺分化期的脑、性腺中gnrh1gnrh2gnrh3的表达水平。结果显示,gnrh1分布于所有检测的组织:在脑、垂体、肾脏和肝脏中高表达,在性腺中表达相对较低;gnrh2则在雌、雄性的肝脏以及雄性的肠、肾脏、眼睛和头肾中;而gnrh3只在雄性的肾脏、眼睛和肌肉中检测到微弱表达。人工诱导的牙鲆雌核发育鱼苗分别用17β-雌二醇(E2)和17α-甲基睾酮(MT)处理,使其分化为雌性或雄性个体表型。在性腺分化期的脑中,gnrh1的表达呈现上升趋势,在全长(total length,TL)4 cm鱼苗中,E2组的表达显著高于MT组(P<0.05);gnrh2在2 cm TL时E2组显著高于MT组(P<0.05),随后在E2和对照组的表达水平均出现下降趋势;gnrh3在2 cm TL时E2和对照组的表达均显著高于MT组(P<0.05),自6 cm TL时MT和E2组的表达开始下降。在性腺中,三种gnrh在性腺分化前2 cm TL表达量都相对较高,随后呈现下降趋势。综上,推测牙鲆gnrh1可能参与了性腺分化的启动、分化过程及性腺发育,gnrh2主要参与了性腺分化的启动,gnrh3主要参与了性腺分化的启动和分化过程。  相似文献   
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