红假单胞菌产类胡萝卜素条件的研究

通过对影响红假单菌类胡萝卜素的因素的全面研究,获得了胡萝卜素产量相对辊主贩培养条件,以乙酸钠为碳源,谷氨酸钠为氮源,酵母亮为生长因子,４ｍｇ／Ｌ,ＭｇＳＯ４,２ｍｇ／ＬｆＥ２（ｓｏ４）３,１５００ｌｘ光照度,３０℃,微好氧培养１２０ｈ。     

Effect of monoclonal antibody to bull sperm sp18 on murine fertilization and embryos development

Western blot analysis revealed that one IgG1 monoclonal antibody (mAb) to sp18 family membrane proteins (Mr. 14, 16 and 18 ku) of bovine sperm reacted faintly with protein bands of 14, 18, 22, 30 and 60 ku (reducing) in samples of mouse sperm. The mAb also reacted to protein of egg lysozyme. Using a laser confocal microscope, indirect immunofluorescence (IIP) showed that the sp18 antigens were present in the posterior head of murine sperm. In murine in vitro fertilization (IVF) and embryo development trails, a total of 426 oocytes from C57BL/6 and F1 hybrid strain (CD1 × C57BL/6 cross) of 12 female mice were used in 3 independent trails. After preincubating capacitated sperm with 182 μg/mL of sp18 mAb in the modified TYH IVF medium for 15-20 min, cumulus-oocyte-complexes were introduced. The fertilization rate in sp18 mAb groups was 77.1 %, which was not significantly (P > 0.05) different from the nonspecific mouse IgG (79.2%) and non-IgG (80.3 %) control groups. Fertilized oocytes had been continuously cultured in modifed CZB medium. 100%, 100% and 97.9% of 1-cell embryos developed to 2-cell stage in sp18 mAb, nonspecific mouse IgG and non-IgG group 30 h after the start of fertilization, respectively. In the nonspecific mouse IgG and non-IgG groups, 64.1 % and 64.3% of embryos developed to the 4-cell stage, respectively, but all developing eggs in sp18 mAb groups arrested development in vitro at 2-cell stage. After zonae of 2-cell blocked embryos were enzy-matically removed with 0.5% pronase, detection of sp18 antigens by IIF indicated that the fluorescence scattered on two embryonic cells. For embryos fertilized in vivo and co-cultured with 182 μg/mL sp18 mAb, the numbers of 1-cell embryos reaching the 2-cell and 4-cell stage were 95. 2% and 70. 5%, which were not significantly (P>0.05) different from the control group (92.9% and 77.9%). These results indicate that the sp18 antigens on posterior head of mouse sperm were incorporated into the egg plasma membrane during fertilization, and played an active role in development of murine preimplanta-tion embryo.     

脉孢霉纤溶酶的酶学性质研究

研究了脉孢霉纤溶酶的部分酶学性质,确定该酶由双亚基组成,亚基分子量分别为30000Da、15500Da,等电点为7.9±0.2,最适作用pH值为7.4,最适作用温度为50℃,在pH6.6～9和32～42℃时较稳定。     

海洋来源真菌的曲酸发酵条件优化

为提高真菌Aspergillus sp.GXIMD02003的曲酸产量,本研究对其利用大米固体培养基发酵的条件进行优化,旨在获得成本低廉的曲酸原料,促进曲酸的工业化生产。研究采用单因素控制变量法考察最佳盐度和最佳发酵时间,通过HPLC法测定曲酸的产量。结果表明真菌Aspergillus sp.GXIMD02003产曲酸的最佳发酵条件为在2%海盐的大米固体培养基中发酵30 d,在此条件下1 000 g大米培养基能够发酵产生24.2 g曲酸。因此,海洋来源真菌Aspergillus sp.GXIMD02003可以作为曲酸的生产菌株,海盐浓度影响该菌株的曲酸代谢。     

不同NaCl含量、培养基和微量元素对分枝杆菌SP-3降解菲的影响研究

研究了分枝杆菌SP 3在不同NaCl含量、培养基和微量元素中降解菲的效果影响。结果表明,分枝杆菌SP 3在NaCl质量分数为1%时对菲的降解率达到最大,为87.41%;菌株在LB液体培养基、改良LB液体培养基和改良的无机盐液体培养基中对菲的降解率都达到了80%以上,三者之间无显著性差异;Fe2+、Zn2+浓度分别在100 μmol/L和5 μmol/L,Mg2+在100 μmol/L时,可以提高菌株对菲的降解能力,而Cu2+随着浓度的增加,能够抑制菌株对菲的降解。研究结果有助于该菌应用于多环芳烃降解的生物技术中,以提高菌株对菲等多环芳烃的降解能力。     

海洋细菌NTa发酵产琼胶酶条件的初步优化

利用选择性培养基从厦门红树林泥土样品中分离得到一株具有较高琼胶酶活力的海洋细菌NTa,通过16S rRNA分析,将该菌株归属为Stenotrophomonas sp.NTa.对该菌株发酵产琼胶酶的条件进行初步优化,结果表明:菌株NTa产琼胶酶的最佳碳源是琼脂,氮源是酵母浸膏,NaCl的质量分数为5%,在28 ℃发酵培养40 h,发酵液的酶活力达到1.98 U/mL,比优化前提高了37.35%.     

Isolation and characterization of a new species of Vibrio-Vibrio olivaceus sp.nov. from diseased flounder （Paralichthys olivaceus temminck et schlegel）

One kind of pathogenic bacteria was isolated from diseased (or dead) flounder (Paralichthys olivaceus L.) which expressed bacterial septicaemia. The phenotyic information of the 10 pure cultures was studied extensively, including morphological characteristics, colony characteristics in different media, physiological and biochemical characteristics, and the mol%G C ratio of the DNA for representative strain. The results showed that the isolates belonged to a new species of Vibrio. In addition, the representative strains have been re-checked by China Center for Type Culture Collection (CCTCC), the results were the same as ours, the examined strains were also regarded as a new species of Vibrio, and designated as Vibrio olivaceus sp.nov. by its isolated fish (Paralichthys olivaceus) based on its biological properties following Rules of International Code of Nomenclature of Bacteria. In the same time, we conducted studies on the serum homology of isolates, pathogenicity of isolates by experimental infection. The results showed that all the isolates were of serologic similarity and the isolates have strong pathogenicity to flounder. Inspection to pure cultures and isolates from liver of moribund fish from infection experiment by fluorescent antibody technique showed the feasibility of fluorescent antibody technique as an auxiliary method of examination.     

鱼腥藻PCC 7120染色体上relBE同源基因的克隆及表达

为研究鱼腥藻PCC7120染色体上与relBE同源的基因asl4561和asl4562表达产物的毒素-抗毒素作用,从鱼腥藻细胞中提取总基因组DNA,设计特异引物PCR扩增目的基因asl4561和asl4562,与T载体连接后经XhoI和EcoR I双酶切并与表达载体pET28a（＋）连接,由IPTG诱导表达,并在IPTG浓度和诱导时间上对asl4561基因的表达条件进行了优化.琼脂糖凝胶电泳显示扩增出了264bp的asl4561基因和213bp的asl4562基因,SDS-PAGE检测表明成功表达出15.65kD和13.55kD的两蛋白,当诱导温度28℃、IPTG浓度0.4 mmol/L、诱导时间6h时,asl4561基因表达蛋白在细菌裂解液上清中表达量最大.     

悬铃木白粉病病原菌及其生物学特性

对近年来在南京、上海、杭州等地危害较为严重的悬铃木白粉病病原菌的无性型和有性型进行了形态学观察和分子生物学鉴定。结果表明,该病害病原的无性型为粉孢属一种(Oidium sp.),病原的有性型为孓蠹菌门的Erysiphe platani。病原菌生长的适宜温度为20~28 ℃,最适温度为26 ℃;湿度对孢子萌发的影响不大,一般环境湿度下都可萌发;分生孢子寿命约为13 d。     

一株高氯酸盐降解菌的分离及特征

利用选择性培养基从ClO4-污染的底泥样品中分离到一株具有ClO4-还原能力的细菌JD15.对其细胞形态、结构、生理特征、16S rDNA和ClO4-还原酶α亚基基因片段pcrA的编码序列分析表明,该菌株为革兰氏阴性的短杆菌,微好氧;16S rDNA序列Dechloromonas sp.SIUL(AF170356.1)的相似性为100%,将其命名为Dechloromonas sp.JD15.该菌株的最适条件为t=25℃,pH=7;能够在ρ(ClO4-)=5×10-3～5×103mg.L-1的培养基上生长;在含0.5～10 mg.L-1的ClO4-培养基中分别培养4 d和30 d后,ClO4-的降解率分别大于50%和80%,表明该菌株可去除环境中的ClO-,具有潜在的应用价值.