鸡白痢沙门氏菌O12特异性噬菌体的分离及生物学特性研究

采用双层平板培养法从成都某养鸡场的污水样本中分离得到一株鸡白痢沙门氏菌O12 (宿主菌)的噬菌体(Ph-1), 并对菌斑形态及理化特性进行了初步研究. 试验结果显示, 纯化后该噬菌体的噬菌斑直径约3mm; 经高于60℃温度处理1h 后噬菌体失活; PH5-7时该噬菌体可保持较高效价和较强的紫外线耐受性; 该噬菌体的最佳感染复数(MOI)为 0.01-0.001; 一步生长曲线显示该噬菌体的潜伏期为40min, 裂解期持续60min.     

鸡白痢沙门氏菌O12特异性噬菌体的分离及生物学特性研究

采用双层平板培养法从成都某养鸡场的污水样本中分离得到一株鸡白痢沙门氏菌O₁₂(宿主菌)的噬菌体(Ph-1), 并对菌斑形态及理化特性进行了初步研究. 试验结果显示, 纯化后该噬菌体的噬菌斑直径约3mm; 经高于60℃温度处理1h 后噬菌体失活; PH5-7时该噬菌体可保持较高效价和较强的紫外线耐受性; 该噬菌体的最佳感染复数(MOI)为 0.01-0.001; 一步生长曲线显示该噬菌体的潜伏期为40min, 裂解期持续60min.     

Construction of heteroduplex DNA and in vitro model for functional analysis of mismatch repair

Functional deficiency of mismatch repair (MMR) system is one of the mechanisms of tumorigenesis. With the development of the investigation and the requirement from the clinical diagnosis and treatment it is necessary to build up a method to evaluate the functional status of the whole MMR system in the concerned tumors. The original ssDNA and dsDNA from wild type (wt) bacteriophage M13mp2 and its three derivates with mutation points in the     

酸碱度对温和噬菌体存活与吸附效率的影响

实验测定了ｐＨ对Ｂａｃｉｌｕｓｔｈｕｒｉｎｇｉｅｎｓｉｓｖａｒ．ｅｎｔｏｍｏｃｉｄｕｓ温和噬菌体ＴＰ３存活与吸附效率的影响作用。结果表明：在不同ｐＨ的介质中,ＴＰ３存活的能力相差很大;酸性介质及高碱性介质对ＴＰ３都有不同程度的瞬间灭活作用;ＴＰ３存活的最适ｐＨ为７．７,且适宜其存活的ｐＨ范围很窄;ｐＨ不同,ＴＰ３吸附到苏云金芽孢杆菌表面上的效率也不同,在ｐＨ≥９范围内,ｐＨ升高,吸附效率降低。     

栖热菌噬菌体TSP4密码子偏嗜性及其对基因异源表达的影响

 为了探索噬菌体TSP4、栖热菌模式菌株Thermus thermophilus HB27中6种蛋白编码序列和Escherichia coli基因组遗传密码子使用偏嗜性,探索TSP4基因密码子偏嗜性对其基因异源表达的影响本研究利用生物学软件Editseq和RSCU算法统计噬菌体TSP4密码子使用频率并分析其偏嗜性,与栖热菌HB27的同源基因以及常温菌E.coli基因组的密码子使用偏嗜性进行对比分析.结果显示,噬菌体TSP4与栖热菌HB27优势密码子相似度高达85%,而与E.coli的相似性仅有65%.为了验证分析结果,选取TSP4基因组中一个潜在分子伴侣基因序列在E.coli BL21和E.coli Rosetta(补充了BL21菌株中缺失的6个稀有密码子)中进行异源表达.噬菌体TSP4与其宿主菌HB27之间密码子高相似性说明在长期的进化过程中TSP4在基因水平上形成对宿主的一种适应性机制.异源表达结果显示,分子伴侣基因在E.coli BL21中未见明显表达,但在Rosetta中高效表达,说明Rosetta中补充的 6个稀有密码子明显有助于分子伴侣基因的表达,该结果也进一步证明密码子偏嗜性是否一致在很大程度上影响基因的表达.     

细菌生物膜与各微生物类群之间的作用关系

从细菌生物膜与真菌、其他细菌、原生动物及噬菌体4个角度对其作用进行了综述。在细菌与真菌之间,生物膜不仅是细菌的拮抗手段,也可介导两者之间的共存。在应对异种细菌时,其作为一种物理防御手段与入侵者竞争。遭受原生动物捕食时,生物膜的致密程度上升以应对原生动物的冲击,或是减少生物膜形成,形成不易被掠食的微菌落。而在抵抗噬菌体侵染方面,生物膜的作用包括使菌体紧密排列,调节本身化学成分变化及群体感应功能,抑制噬菌体感染。     

236例儿科抗菌素使用对比分析

目的为了解儿科抗菌素应用的变化及现状,探讨改善儿科抗菌素滥用趋势的途径。方法分别随机抽取时间跨度10年两组儿科住院病历,对其诊断,用药指征、用药时间、用药种类、用药量进行对比分析。结果住院病例全部使用抗菌素、X线、实验室检查支持使用抗菌素分别为75%和47.1%,抗菌素越用越新,用量偏大。结论必须建立系统的到位的干预措施,强化继续教育,加强知识更新和病原微生物的临床研究。     

用PCR法检测苏云金杆菌的溶原性噬菌体

根据苏云金杆菌(Bacillus thuringiensis,B.t)溶原性噬菌体G IL01(GenBank Accession Number:AJ536703)的同源序列设计一对引物,用35株B.t标准菌株和生产菌株MZ1(血清型H3 a3b)的过夜培养物作模板,结果有16株菌株包括MZ1能扩增出大小约为750 bp的预期产物,推测这16株菌为溶原菌。MZ1菌株的过夜培养液与指示菌ZK1(血清型H25)混匀并在双层琼脂上培养12 h后出现了滴度为2×107pfu的噬菌斑,从而证明生产菌株MZ1确为溶原菌;又以G IL01同源性较高的其它四个基因设计引物,分别以MZ1的培养物及其溶原性噬菌体基因组为模板进行PCR扩增,结果两者得到几乎一致的带型,进一步证明了生产菌株MZ1为溶原菌,说明利用PCR法检测鉴定苏云金杆菌的溶原性是可行的。比较指示菌方法、直接电镜法和DNA鉴定法,PCR法快速简便,对B.t生产上快速检测其溶原性噬菌体有实用意义。     

Construction of heteroduplex DNA and<Emphasis Type="Italic">in vitro</Emphasis> model for functional analysis of mismatch repair

Functional deficiency of mismatch repair (MMR) system is one of the mechanisms of tumorigenesis. With the development of the investigation and the requirement from the clinical diagnosis and treatment it is necessary to build up a method to evaluate the functional status of the whole MMR system in the concerned tumors. The original ssDNA and dsDNA from wild type (wt) bacteriophage M13mp2 and its three derivates with mutation points in the lacZα gene have been used to construct two kinds of hetero-duplex DNA molecules. One named del(2) has two bases deleted in the negative strand, the other has a G·G mismatch base pair in the negative strand too. Introducing this heteroduplex DNA into E. coli NR9162 (mutS-) without the MMR ability on the indicator plate with x-gal and IPTG, there are three kinds of plaques, mixture plaque as the characteristic phenotype of heteroduplex DNA, blue and clear plaques. If the cell extract is mismatch repair competent the percentage of the mixture plaque will decrease after incubation with these heteroduplex DNA, the repair efficiency is expressed in percentage as 100× (1 minus the ratio of percentages of mixture plaque obtained from the extract-treated sample and untreated samples), which can imply the functional status of MMR system of certain samples. After large T-antigen-dependent SV-40 DNA replication assay cell extract from TK6, a human lymphoblastoid B-cell lymphoma cell line with MMR ability, and Lovo, a human colonic carcinoma cell line with MMR deficiency have incubated with these heteroduplex DNA. The repair efficiency of TK6 to del(2) is more than 60%, to G-G is more than 50%. The Lovo efficiency to del(2) is less than 10%, to G-G is less than 20%. Therefore, in this in vitro model used for functional analysis of mismatch repair of heteroduplex DNA as the repair target, TK6 can serve as the control for MMR proficiency and Lovo as the control for MMR deficiency. Using this model the tumor tissue from a case of hereditary nonpolyposis colorectal cancer (microsatellite instability high, MSI-H) was measured and lack of MMR ability was shown. And a case of sporadic rectal cancer (SRC) (microsatellite stability, MSS) maintains MMR proficiency. The results indicate that the model is sensitive and dependable. It could be used to measure the func- tion status of MMR system in tumor cell and/or tissues. This is a reliable method to investigate the mechanic of tumori-genesis. It is meaningful in the observation of the role of MMR in the initiation and progression of concerned tumors.     

病毒作为纳米模板的研究进展

随着基因工程和纳米技术的发展,病毒的材料学应用潜能逐渐彰显.根据材料科学的观点,病毒可被看作有机纳米颗粒,其大小、形态以及表面的功能基团的数量和性质都是经过精确定义的.由于具有合适的大小、形状和明确的化学结构,病毒可被用作材料组装的纳米量级模板,调节无机材料、金属材料、磁性材料、半导体材料等的生长和成型.本文主要以丝状结构的M13噬菌体和球状结构的豇豆花叶病毒为例,阐述病毒作为模板在纳米导线、纳米电池、晶体管、存储器电路等领域的应用进展.