亚硝酸盐影响Lactobacillus brevis 4903发酵的研究

通过研究可知,亚硝酸盐对Lactobacillusbrevis4903发酵有抑制作用,环境中亚硝酸盐一旦分解掉,这种抑制作用就会被解除。分析其原因:①亚硝酸盐抑制了乳酸菌生长,从而抑制了乳酸发酵;②在发酵初期可能因亚硝酸盐还原酶的作用,使亚硝酸盐酶解生成NH3,NH3中和了乳酸菌生成的酸(H ),从而使环境pH值的下降和酸的积累变得缓慢。     

蛋白酶抗性人睫状神经营养因子突变体基因的构建及其在巴斯德毕赤酵母中的表达

AX15是一种比天然睫状神经营养因子具有更高的生物学活性、更好的稳定性和可溶性的hCNTF突变体。在巴斯德毕赤酵母中表达时AX15易发生降解。氨基酸序列分析表明降解位于由12和13位氨基酸残基组成的双碱性氨基酸之后。根据KEX2蛋白酶的底物专一性把双碱性氨基酸从RR变为RK,构建了KEX2抗性的AX15突变体AX15（R13K）。AX15（R13K）的稳定性得到了显著的提高,在诱导100 h后也未发生降解。利用超滤浓缩和凝胶过滤得到了纯度>90%的AX15（R13K）。TF-1细胞存活实验表明AX15（R13K）具有与AX15相同的生物学活性。蛋白酶抗性人睫状神经营养因子突变体可能具有更好的体内稳定性,在临床应用上具有潜在的优势。     

系统生物学——生命科学的新领域

系统生物学是继基因组学、蛋白质组学之后一门新兴的生物学交叉学科,代表21世纪生物学的未来.最近,系统生物学研究机构纷纷成立.在研究上,了解一个复杂的生物系统需要整合实验和计算方法.基因组学和蛋白质组学中的高通量方法为系统生物学发展提供了大量的数据.计算生物学通过数据处理、模型构建和理论分析,成为系统生物学发展的一个必不可缺、强有力的工具.在应用上,系统生物学代表新一代医药开发和疾病防治的方向.     

重组小鼠canstatin N端片段对鸡胚新生血管生成的抑制作用(英)

为了研究重组小鼠canstatin N端片段的体内抗血管生成活性, 通过PCR扩增小鼠canstatin N端片段cDNA,定向克隆于原核表达载体pET30a(+)中,构建小鼠canstatin N端片段重组表达载体pET-mCanN, 转化E.coli BL21(DE3), IPTG诱导表达,SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹检测小鼠canstatin N端片段的表达. 结果表明,IPTG诱导原核表达载体pET-mCanN在大肠杆菌E.coli BL21(DE3)中高效表达, 小鼠canstatin N端片段表达量约占菌体总蛋白量的18%, 小鼠canstatin N端片段主要以包涵体形式存在,包涵体经过洗涤、裂解、Ni-spin column亲合柱层析以及蛋白质复性等步骤纯化后,获得了纯度约为92%的重组小鼠canstatin N端片段. 鸡胚绒毛尿囊膜(chicken embryo choriollantoic membrane,CAM)实验表明,原核表达的小鼠canstatin N端片段能有效地按剂量依赖的方式抑制鸡胚新生血管的形成.     

引种台湾桤木的水分生理特性

台湾桤木(Alnusformosana)原产台湾,是台湾地区重要的先锋造林树种和林地培肥树种。水分是限制植物地理分布和决定植物引种能否成功的重要因子之一。采用Li-6400光合测定系统对引种两年生台湾桤木苗木的水分生理特性进行了研究,结果表明:(1)在整个生长期,叶片蒸腾速率日变化均表现为宽大的单峰型曲线,以13∶00～15∶00时左右蒸腾速率最高,不同部位叶片在不同生长期的峰值有所不同,属非蒸腾午休型植物。气孔导度日变化有单峰型和双峰型2种变化曲线;(2)蒸腾速率季节变化表现为单峰曲线,以7月为最高,整个生长期叶片日均蒸腾速率为3.25mmol·m-2·s-1,表现出高光合、强蒸腾的生理特点;(3)水分利用效率日变化与光合速率日变化相似,有单峰曲线和双峰曲线2种类型,与气孔导度的日变化有显著相关性。在整个生长期,以6月、7月叶片水分利用效率为最低,8月为最高;(4)在影响水分利用效率季节变化的生理生态因子中,以温度、光合有效辐射、气孔导度和相对湿度4个因子的直接作用为最大。     

偏分离分子标记的作图方法

对取自MAPMAKER软件小鼠F2群体(含333个体)的5个RFLP连锁标记数据作了共显性分子标记偏分离的分析。先确定选择类型的方程组(配子或合子),随后采用Newton-Raphson迭代法估算标记间的重组值。在构建分子标记遗传图谱时,如果两个相邻标记均存在偏分离,最好采用纳入偏分离因子的估算方法。在估计F2群体标记间偏分离重组距离上,用连续χ2检测方法比传统χ2检测更为准确。Abstract The comparative analysis of segregation distortions of the codominant markers data presented in software MAPMAKER are made, where five RFLPs markers involve in a mouse F2 population with 333 individuals. The successive χ2 test begins with the determinations of gametic or zygotic selection types, followed by the estimation of recombination fractions between two markers with the Newton-Raphson iteration method. It is better to use the molecular marker showing segregation distortion for constructing a genetic map, in the case of seriously skew segregation between both the adjoining markers. The successive χ2 test provides better accuracy than that of classical χ2 test for the estimation of the recombination values in F2 population with segregation distortion.     

偏分离分子标记的作图方法

对取自MAPMAKER软件小鼠F_2群体(含333个体)的5个RFLP连锁标记数据作了共显性分子标记偏分离的分析。先确定选择类型的方程组(配子或合子),随后采用Newton-Raphson迭代法估算标记间的重组值。在构建分子标记遗传图谱时,如果两个相邻标记均存在偏分离,最好采用纳入偏分离因子的估算方法。在估计F_2群体标记间偏分离重组距离上,用连续x~2检测方法比传统x~2检测更为准确。     

植物蛋白质组学研究进展

植物结构蛋白质组学研究目的在不断变化。早期旨在比较不同基因型和株系,估测系统发育距离。随后是用蛋白N端Edman测序法或氨基酸分析法鉴定蛋白。如今,应用双向凝胶电泳和质谱法,已能联手成功地解决蛋白分离和鉴定问题。这将有助于对不同突变系与野生型作比较和对不同的表达基因鉴定,应用数据库和作图软件有可能获得功能蛋白构象。     

Quantitative real-time PCR assay to detect transgene copy number in cotton (Gossypium hirsutum)

A TaqMan quantitative real-time PCR detection system was developed to examine transgene copy number in cotton. GhUBC1, a gene validated to be present as a single copy per haploid Gossypium hirsutum genome, was used as the endogenous reference to estimate copy number of GFP and selection marker NPTII in 28 T0 plants. This system was found to be more accurate than genomic Southern blot hybridization and could effectively tell homozygotes from heterozygotes in a T1 transgenic cotton population. Therefore it is suitable for efficient and cost effective early screening of transgenic seedlings and identifying transgene homozygotes in segregation populations.     

Characterization of DegQVh, a Serine Protease and a Protective Immunogen from a Pathogenic Vibrio harveyi Strain

Vibrio harveyi is an important marine pathogen that can infect a number of aquaculture species. V. harveyi degQ (degQ_Vh), the gene encoding a DegQ homologue, was cloned from T4, a pathogenic V. harveyi strain isolated from diseased fish. DegQ_Vh was closely related to the HtrA family members identified in other Vibrio species and could complement the temperature-sensitive phenotype of an Escherichia coli strain defective in degP. Expression of degQ_Vh in T4 was modulated by temperature, possibly through the σ^E-like factor. Enzymatic analyses demonstrated that the recombinant DegQ_Vh protein expressed in and purified from E. coli was an active serine protease whose activity required the integrity of the catalytic site and the PDZ domains. The optimal temperature and pH of the recombinant DegQ_Vh protein were 50°C and pH 8.0. A vaccination study indicated that the purified recombinant DegQ_Vh was a protective immunogen that could confer protection upon fish against infection by V. harveyi. In order to improve the efficiency of DegQ_Vh as a vaccine, a genetic construct in the form of the plasmid pAQ1 was built, in which the DNA encoding the processed DegQ_Vh protein was fused with the DNA encoding the secretion region of AgaV, an extracellular β-agarase. The E. coli strain harboring pAQ1 could express and secrete the chimeric DegQ_Vh protein into the culture supernatant. Vaccination of fish with viable E. coli expressing chimeric degQ_Vh significantly (P < 0.001) enhanced the survival of fish against V. harveyi challenge, which was possibly due to the relatively prolonged exposure of the immune system to the recombinant antigen produced constitutively, albeit at a gradually decreasing level, by the carrier strain.