PCR-SSP技术对广东汉族人HLA-DR基因分型

探索具有高分辨率、高特异性和简捷快速的方法对ＨＬＡ－ＤＲ基因分型,为临床器官移植配型和疾病相关性分析提供实用的方法和基础资料．利用ＤＲ１～ＤＲｗ１８序列特异性的１９组引物及１对内参照引物进行ＰＣＲ扩增即ＰＣＲ－ＳＳＰ对ＨＬＡ－ＤＲ进行基因分型,扩增产物经琼脂糖凝胶电泳,溴乙锭染色,在紫外光下观察分型结果．每个被检个体的ＤＲ型别可由特异引物扩增出现的电泳谱带直接判断．双盲检测２２例的结果１００％正确．在１０２例中国广东地区汉族人中,ＤＲ９和ＤＲ２的基因频率最高,分别为０．２２０５和０．１９１２,ＤＲ１０为最低（０．００９８）．与用ＰＣＲ－ＳＳＯ方法分型获得的结果比较,基因型别分布基本一致,但一些等位基因的频率有差异,表明ＨＬＡ－ＤＲ基因频率的分布在不同地区、不同种族的人群间存在着差异．ＰＣＲ－ＳＳＰ法分辨率和特异性虽不及ＰＣＲ－ＳＳＯ法但比血清学方法精细,分型的全过程只需２～４ｈ能满足临床器官移植配型的要求．基因频率调查结果为器官移植配型和疾病相关性分析提供了基础资料．     

紫外照射对DNA修复基因RAD24转录的影响

以酿酒酵母HY684为实验对象,用波长254nm紫外线分别在0,37,50和70J/m~2等强度下照射不同时间,提取总RNA,应用North-ern杂交方法检测RAD24基因转录水平的变化。结果显示37J/m~2、50J/m~2照射20分钟到120分钟明显提高RAD24基因转录水平,70J/m~2照射时,则恢复至正常水平,这说明低剂量紫外线照射可提高RAD24基因转录水平,该基因的表达具有损伤诱导性。     

Anti-AIDS agents 82: Synthesis of seco-(3′R,4′R)-3′,4′-di-O-(S)-camphanoyl-(+)-cis-khellactone (DCK) derivatives as novel anti-HIV agents

Thirteen novel seco-DCK analogs (4–16) with several new skeletons were designed, synthesized and screened for in vitro anti-HIV-1 activity. Among them, three compounds (5, 13, and 16) showed moderate activity, and compound 9 exhibited the best activity with an EC₅₀ value of 0.058 μM and a therapeutic index (TI) of 1000. The activity of 9 was better than that of 4-methyl DCK (2, EC₅₀: 0.126 μM, TI: 301.2) in the same assay. Additionally, 9 also showed antiviral activity against a multi-RT inhibitor-resistant strain (RTMDR), which is insensitive to most DCK analogs. Compared with 2, compound 9 has a less complex structure, fewer hydrogen-bond acceptors, and a reduced log P value. Therefore, it is likely to exhibit better ADME, and appears to be a promising new lead for further development as an anti-HIV candidate.     

枫香叶片变色期全长转录组测序及分析

枫香因其树形优美,入秋后叶色红艳或橙黄,极具观赏价值,是优良的景观生态树种。为了解枫香叶片变色及其次级代谢过程的遗传基础,该文以枫香5个叶片变色期叶片混合样品为材料,利用单分子实时测序技术(PacBio平台)对其进行全长转录组测序。结果表明:(1)全长转录组测序共获得41.04 Gb的高质量数据,从中鉴定出全长非嵌合序列563 180条,通过聚类和去冗余,获得27 269条高质量全长转录本。在27 269条全长转录本中预测到2 035条长链非编码RNA(lncRNA),并检测出14 892个简单重复序列(SSR)位点和1 856个转录因子。(2)基因注释结果表明,NR、GO、COG、KEGG 等8个数据库共注释了24 857条转录本,KEGG数据库共获得了124个条代谢途径,主要有核糖体、碳代谢、氨基酸生物合成等,在类黄酮和叶绿素代谢途径中分别有49和71个转录本参与。上述结果初步揭示了枫香叶片变色期转录组信息以及功能特性,为后续研究枫香叶片变色分子机制、色素代谢合成途径和调控、相关功能基因克隆以及叶色改良提供基础数据。     

Synthesis and DNA cleavage activity of 2-hydrazinyl-1,4,5,6-tetrahydropyrimidine containing hydroxy group

2-Hydrazinyl-1,4,5,6-tetrahydropyrimidin-5-ol dihydrochloride 2, as well as 2-hydrazinyl-4,5-dihydro-1H-imidazole dihydrochloride 1, was synthesized as metal-free DNA cleaving agent. Agarose gel electrophoresis was used to assess the plasmid pUC 19 DNA cleavage activities in the presence of 1 and 2. DNA cleavage efficiency of 2 exhibits remarkable increases compared with its corresponding non-hydroxy compound 1. Kinetic data of DNA cleavage promoted by 2 fit to the Michaelis–Menten-type equation with k_max of 0.0378 ± 0.0013 h^?1 giving 10⁶-fold rate acceleration over uncatalyzed DNA. The acceleration is driven by the spatial proximity of the nucleophilic hydroxy group and the electrophilic activation for the phosphodiester by the ammonium and/or guanidinium groups. In vitro cytotoxic activities toward Hela cells and human leukemia HL-60 cells were also examined, and 2 exhibits stronger cytotoxic activities than 1.     

Alkali‐catalyzed low temperature wet crosslinking of plant proteins using carboxylic acids

We report the development of a new method of alkali‐catalyzed low temperature wet crosslinking of plant proteins to improve their breaking tenacity without using high temperatures or phosphorus‐containing catalysts used in conventional poly(carboxylic acid) crosslinking of cellulose and proteins. Carboxylic acids are preferred over aldehyde‐containing crosslinkers for crosslinking proteins and cellulose because of their low toxicity and cost and ability to improve the desired properties of the materials. However, current knowledge in carboxylic acid crosslinking of proteins and cellulose requires the use of carboxylic acids with at least three carboxylic groups, toxic phosphorous‐containing catalysts and curing at high temperatures (150–185°C). The use of high temperatures and low pH in conventional carboxylic acid crosslinking has been reported to cause substantial strength loss and/or undesired changes in the properties of the crosslinked materials. In this research, gliadin, soyprotein, and zein fibers have been crosslinked with malic acid, citric acid, and butanetetracarboxylic acid to improve the tenacity of the fibers without using high temperatures and phosphorus‐containing catalysts. The new method of wet crosslinking using carboxylic acids containing two or more carboxylic groups will be useful to crosslink proteins for various industrial applications. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009