去甲基万古霉素菌种低温冷冻、冷干保藏条件优化及10 年菌种保藏效果

【目的】针对去甲基万古霉素产生菌不耐保藏的问题,改进菌种保藏方法,对超低温液氮保藏、-80°C低温冷冻保藏、冷干保藏方法跟踪考察10年保藏稳定性,评价不同保藏方法对去甲基万古霉素产生菌的保藏适用性。【方法】采用甘油作基础保护剂进行超低温液氮保藏和-80°C低温冷冻保藏,采用脱脂牛奶作基础保护剂进行冷干保藏,针对超低温液氮保藏进行降温速率考察,研究非渗透性冷冻保护剂海藻糖、聚乙烯吡咯烷酮(PVP)等对3种保藏方法的冻存影响,对优选出的保藏方法进行10年跟踪考察。【结果】3种保藏方法冻后菌种存活率依次为:-80°C低温冷冻保藏超低温液氮保藏冷干保藏。液氮保藏最适降温速率为快速冷冻。优选出最佳保护剂配方:超低温液氮保藏为甘油8.0%,海藻糖3.5%;-80°C低温冷冻保藏为甘油6.0%,PVP 5.0%;冷干保藏为脱脂牛奶,6.0%海藻糖。采用优化保藏条件,液氮保藏10年存活率稳定在70.6%,菌种发酵水平为入藏水平的92.9%。【结论】在优化条件下,尤以超低温液氮保藏适合于去甲基万古霉素产生菌长期保藏。     

Comparison of Ethanol Production by Different Zymomonas Strains

A comparison of the rates of growth and ethanol production by 11 different strains of Zymomonas revealed a wide range of characteristics, with some strains being more tolerant of high sugar or ethanol concentrations and high incubation temperatures than others. Some strains were unable to utilize sucrose; others produced large amounts of levan, and one strain grew well but produced no levan. One strain, CP4, was considerably better in all respects than most of the other strains and was chosen as a starting strain for genetic improvement of ethanol production.     

Comparison of beta-Glucosidase Activities in Different Streptomyces Strains

Cellobiase (beta-glucosidase) production was compared for two streptomycetes: Streptomyces flavogriseus, a known producer of cellulase complex, and Streptomyces sp. strain CB-12, a strain isolated for its rapid growth on cellobiose. The optimal conditions for enzyme activity were established in relation to pH, temperature, enzyme stability, and substrate affinity. The production of beta-glucosidase by the two strains depended on the carbon substrate in the medium. Cellobiose was found to repress the biosynthesis of the enzyme in S. flavogriseus and to stimulate its production in strain CB-12. The biosynthesis of the enzyme correlated well with the accumulation of glucose in the culture filtrates. The combined action of the beta-glucosidases produced by the two Streptomyces strains might allow a better utilization of the reaction products which arise during the biodegradation of cellulose.     

红曲霉不同转化方法比较

为了研究红曲霉聚酮体途径,考察和比较了4种不同的转化方法以建立有效的红曲霉遗传转化系统。以潮霉素作为抗性筛选标记,pBC-Hygro作为转化载体,用基于原生质体的传统转化和电击转化、基于萌发孢子的电击转化以及REMI技术转化红曲霉。发现基于萌发孢子的电击转化由于转化率极低而不适于红曲霉转化。基于原生质体的传统转化和电击转化尽管每微克DNA分别能获得135个转化子和125个转化子,但因转化子稳定性差也适合红曲霉转化的转化。应用REMI技术,转化率提高约20倍,每微克DNA 2500个转化子,70%～75%的转化子的稳定,非常适合于红曲霉的转化。      

生物破乳剂产生菌的筛选及其方法研究

针对生物破乳剂产生菌筛选难的问题,采用显色法、溶血细胞测试法、表面张力测定法和排油圈法从6种不同菌源对生物破乳菌产生茵进行了筛选.通过试验筛选得到了17株生物破乳剂产生茵,其中24h内破乳率高于70%的破乳菌有5株;油田含油污泥、采油废水生物处理污泥和污水处理厂剩余污泥是筛选破乳菌的较好的菌源:显色法、溶血圈法存在检测范围的局限性;表面张力测定法和排油圈法是最为简易和准确的生物表面活性剂产生茵的筛选方法,采用模型乳状液对生物破乳剂产生菌进行筛选最为直接和准确,但工作量大、所需时间长,因此在筛选高效破乳菌时,建议采用表面张力、排油圈法进行初筛,而后通过模型乳状液破乳进行验证.     

噬菌体浓缩方法的比较

目的：对不同噬菌体浓缩方法进行比较。方法：采用PEG沉淀、PEG反透析、阴阳离子结合树脂、超滤等5种方法对T4、T7、N4等3类噬菌体进行浓缩对比试验;在此基础上,用浓缩效果较好的方法对土壤和河道污水样本中的噬菌体进行浓缩,观察浓缩效果。结果：上述5种方法对3类噬菌体均有浓缩效果;超滤的浓缩效果最好,其次是PEG反透析,PEG沉淀没有很好的浓缩效果;河水样本中噬菌体的回收率比土壤样本高。结论：可通过PEG反透析、超滤或两者相结合的方法,得到高浓度的有活性的噬菌体。     

生物破乳剂产生菌的筛选及其方法研究

针对生物破乳剂产生菌筛选难的问题, 采用显色法、溶血细胞测试法、表面张力测定法和排油圈法从6种不同菌源对生物破乳菌产生菌进行了筛选。通过试验筛选得到了17株生物破乳剂产生菌, 其中24h内破乳率高于70%的破乳菌有5株; 油田含油污泥、采油废水生物处理污泥和污水处理厂剩余污泥是筛选破乳菌的较好的菌源; 显色法、溶血圈法存在检测范围的局限性; 表面张力测定法和排油圈法是最为简易和准确的生物表面活性剂产生菌的筛选方法, 采用模型乳状液对生物破乳剂产生菌进行筛选最为直接和准确, 但工作量大、所需时间长, 因此在筛选高效破乳菌时, 建议采用表面张力、排油圈法进行初筛, 而后通过模型乳状液破乳进行验证。     

Comparison of Genomes in Streptococcus thermophilus Strains of Different Origin

According to DNA hybridization data, thermophilic streptococci used in Russia as starters in the dairy industry are divided into six different genomovars, with a degree of DNA homology not exceeding 20–50%. The analysis of genomes from these genomovars using SmaI restriction endonuclease and pulsed-field gel electrophoresis revealed wide variability of the genome size. In some strains, the genome size considerably exceeded 2000 kbp. Most of the strains studied contained plasmids about 120 kbp in size.     

Comparison of Different Methods for Preparation of Human Milk Casein

Changes of the estradiol and lipid concentrations in hen serum, and those in the fatty acids of hen and cockerel livers were studied during the maturing period. The concentrations of estradiol and lipid in serum reached a maximum level a few days before the onset of laying, and then fell. A marked increase in oleate, and decreases in stearate and arachidonate were observed in both the liver and serum lipids of the hen when reaching laying. The fatty acid compositions of the serum and liver of the hen at 145 and 167 days of age were close to those of yolk lipid. In the cockerel, high contents of stearate and arachidonate and a low content of oleate were observed, and the fatty acid composition did not change appreciably with age. On estrogenization of the male chick, the fatty acid cmposition of the liver lipid was similar to that of the laying hen liver. The ratio of the unsaturated fatty acid/saturated fatty acid in the liver of hen increased when reaching laying, the value at 167 days old being close to that of yolk lipid. This ratio in the liver lipid of cockerel did not change significantly throughout the experimental period.     

啮齿类螺杆菌不同检测方法的比较

目的确定用于实验啮齿类动物常规螺杆菌检测方法。方法选用针对螺杆菌属16S rRNA属特异性的4对引物进行了引物选择、取材部位以及与分离培养法比较。结果引物P7/P8的敏感性优于其它3对,检出限可达0.01 pg,对阳性动物群检出率80%-100%;分离培养法检出率40%-50%;盲肠、结肠检出率无显著差异。结论以引物P7/P8的PCR方法作为啮齿类实验动物螺杆菌初筛方法,分离培养法作为验证方法,取材部位可在盲肠或结肠。     

苯丙酮尿症常用筛查方法的比较

苯丙酮尿症是一种由于体内缺乏苯丙氨酸羟化酶所引起的遗传性代谢异常疾病,如果能够做到早诊断、早发现、早预防和早治疗,就可以避免患儿以后出现智能发育障碍,因此开展新生儿的筛查工作十分重要。简要介绍了苯丙酮尿症的基本情况以及目前应用于临床的常用筛查方法,如细菌抑制法、荧光法等,并从筛查原理、主要优缺点等方面进行了简单的比较。