农杆菌介导的大麦Mlo反义基因转化小麦获得抗白粉病后代

从大麦‘斯特林’幼叶总RNA中分离Mlo基因cDNA完整编码区,反向连接到植物双元载体（pBI-121.2）35S启动子下游,通过农杆菌介导的苗端转化法获得两种小麦基因型（‘烟优2801’和‘烟优361’）的转基因小麦。T0代405株中有55株PCR检测阳性,平均转化率达到13.58%,T0和T1基因组DNA Southern杂交可以证明大麦Mlo基因片段已整合到小麦基因组中并可传递到后代。两种基因型的转基因小麦T0和T1植株在温室及大田中均表现出对白粉病抗性的提高。农杆菌介导的苗端转化法可以简单、快速、高效地获得转基因株系;排除体细胞变异对转基因植株的影响;克服基因型对农杆菌转化的限制,是小麦遗传转化的一种实用方法。     

Notch signaling maintains proliferation and survival of the HL60 human promyelocytic leukemia cell line and promotes the phosphorylation of the Rb protein

The Notch signaling pathway has been implicated in the development of several leukemia and lymphoma. In order to investigate the relationship between Notch signaling and acute myeloid leukemia (AML), in this study, we expressed a recombinant Notch ligand protein, the DSL domain of the human Jagged1 fused with GST (GST-Jag1). GST-Jag1 could activate Notch signaling in the human promyelocytic leukemia cell line HL60, as shown by a reporter assay and the induced expression of Notch effector gene Hes1 and Hes5. However, GST-Jag1 had no effect on the proliferation and survival of HL60 cells. HL60 cells expressed both Notch ligands and receptors, and had a potential of reciprocal stimulation of Notch signaling between cells. We, therefore, blocked Notch signaling in cultured HL60 cells using a γ-secretase inhibitor (GSI). We found that GSI inhibited the proliferation of HL60 cells significantly by blocking the cell-cycle progression in the G1 phase. Furthermore, GSI induced remarkably apoptosis of HL60 cells. These changes in GSI-treated HL60 cells correlated with the down-regulation of c-Myc and Bcl2, and the low phosphorylation of the Rb protein. These results suggested that reciprocal Notch signaling might be necessary for the proliferation and survival of AML cells, possibly through the maintenance of the expression of c-Myc and Bcl2, as well as the phosphorylation of the Rb protein.     

Natural selection maintains the transcribed LTR retrotransposons in Nosema bombycis

Eight intact LTR retrotransposons(Nbr1?Nbr8)have been previously characterized from the genome of Nosema bombycis,a eu-karyotic parasite with a compact and reduced genome.Here we describe six novel transcribed Nbr elements(Nbr9?Nbr14)identified through either cDNA library or RT-PCR.Like previously determined ones,all of them belong to the Ty3/Gypsy superfamily.Retrotransposon diversity and incomplete domains with insertions(Nbr12),deletions(Nbr11)and in-frame stop codons in coding regions(Nbr9)were detected...     

应用球面对称设计方法优化谷胱甘肽生物合成条件

研究了提高细胞内谷胱甘肽质量分数的方法。谷胱甘肽的总产量与细胞的数量和细胞内谷胱甘肽质量分数有关,通过添加底物和刺激物可以促进谷胱甘肽生物合成,提高胞内谷胱甘肽的质量分数。实验考察了高渗刺激物与前体氨基酸对胞内谷胱甘肽质量分数的影响,并应用球面对称设计优化了实验条件,使得胞内谷胱甘肽质量分数达4.47%,产量达257.3mg.L-1,分别比优化前提高了约69.3%和75.7%。     

大肠杆菌RecQ解旋酶的表达纯化和生物学活性研究

RecQ家族解旋酶是DNA解旋酶中高度保守的一个重要家族,在维持染色体的稳定性中起着重要的作用.人类RecQ家族解旋酶突变会导致几种与癌症有关的疾病.本研究旨在诱导大肠杆菌RecQ解旋酶体外表达,并应用生物化学和生物物理学技术研究大肠杆菌RecQ解旋酶的生物学活性. 体外诱导表达获得纯度达90% 以上并具有高活性的大肠杆菌重组RecQ解旋酶,其可溶性好;经生物学活性分析显示具有DNA结合活性、ATP依赖的DNA解链活性、DNA依赖的ATP酶活性. 较之双链DNA（dsDNA）,大肠杆菌RecQ解旋酶更容易与单链DNA(ssDNA)结合( P<0.01 ),但与长度不同的dsDNA的结合特性有差异(P<0.01)而与ssDNA没有差异(P>0.05);大肠杆菌RecQ解旋酶对3种dsDNA的解链速率不同(P<0.05);大肠杆菌RecQ解旋酶的ATP酶活性与辅助因子ssDNA长度也呈正相关(P<0.01). 这些研究结果将有助于阐明大肠杆菌RecQ解旋酶的分子作用机制,并为研究RecQ解旋酶家族其它成员的结构与功能提供帮助.     

The In vitro antioxidant properties of chinese highland lichens

Antioxidant properties of 46 lichen species collected from high-UV exposed alpine areas of southwestern China were evaluated for their therapeutic utilization. Anti-linoleic acid peroxidation activity, 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging activity, reducing power and the total phenolic contents were assessed in methanol extract of the lichens in vitro. Extracts of Peltigera praetextata and Sticta nylanderiana exhibited potent activity in all antioxidant tests. Especially, extracts of S. nylanderiana exhibited 1.37 times higher anti-linoleic acid peroxidation activity than ascorbic acid used as a positive control. It also showed the strongest free radical scavenging activity among all the tested species with an inhibition of 90.4% at the concentration of 330microgram/ml. Potent reducing power was also detected in the lichen extract compared with BHA. Generally, the antioxidant lichens contained large amount of phenolic contents. The activity-guided bioautographic TLC and HPLC analysis are used to find out the compounds responsible for the potent antioxidant activities of S. nylanderiana extract. The analysis demonstrated that lecanoric acid is one of the effective antioxidant compounds in S. nylanderiana. The results suggested that several highland lichens can be used as a novel bioresource for natural antioxidants.     

细胞角蛋白19启动子调控的双报告载体的构建及其在肝干/祖细胞分化研究中的应用

肝干/祖细胞是肝细胞和胆管上皮细胞(biliary epithelial cells,BECs)共同的前体细胞,为了对这一前体细胞的分化情况进行研究,利用报告基因来监测肝干/祖细胞的分化走向．首先,通过PCR方法从肝癌细胞系HepG2的全基因组中克隆了细胞角蛋白19(CK19)启动子片段,构建了CK19启动子调控的海肾荧光素酶和红色荧光蛋白(red fluorescent protein,RFP)双报告载体(pSicoR-CK19-hrl-mrfp)．其次,将上述慢病毒载体转染肝干/祖细胞后,通过流式细胞分选获得稳定转染的细胞株．再次,将上述细胞株与表达“上皮形态发生素”(Epimorphin,EPM)的PT67细胞共培养后,整合有pSicoR-CK19-hrl-mrfp表达载体的肝干/祖细胞不仅形态发生了变化,而且排列为二维环状结构,另外还检测到由CK19启动子启动表达的海肾荧光素酶和RFP,细胞形态和基因表型都证明肝干/祖细胞经诱导已经分化成为BECs．与此形成对照的是肝干/祖细胞与不表达EPM的PT67细胞共培养后,没有观测到上述的变化．所以,CK19启动子调控的双报告载体不仅可以实时地显示肝原始细胞在不同的诱导环境下的分化走向,而且还可以定量地检测CK19启动子活性的变化情况．总之,这一载体的成功构建将为研究肝干/祖细胞的分化提供了便捷的工具,同时也有助于筛选可诱导肝干/祖细胞定向分化的分子．     

大山包黑颈鹤自然保护区亚高山植物区系

在野外调查和标本鉴定的基础上,对大山包黑颈鹤国家级自然保护区维管植物多样性进行了统计分析,结果表明:该地区植物种类丰富,有维管植物72科197属358种(亚种和变种),其中栽培植物18种、外来入侵植物3种.其植物区系性质为温带性,为东亚地区高寒山地植物区系.大山包现代植物区系缺乏裸子植物和乔木树种归因于人类长期活动的干扰,水生植物种类贫乏是自然规律.     

Oligonucleotide-selenide conjugate: Synthesis and its inducible sequence-specific alkylation of DNA

Oligonucleotide-selenium conjugate was designed and synthesized and its sequence-specific cross-linking ability was investigated. The selenide derivatives can generate covalent interstrand cross-linking with its complementary strand through the formation of o-QM intermediate induced by periodate oxidation. A cross-linking reaction yield of up to 50% was obtained. Hydroxyl radical footprinting experiment revealed that the quinone appendage specifically alkylated the cytosine base extending the duplex formed between the conjugate and the target strand.     

2,3-Diamino acid modifying 3S-tetrahydroisoquinoline-3-carboxylic acids: Leading to a class of novel agents with highly unfolded conformation,selective in vitro anti-platelet aggregation and potent in vivo anti-thrombotic activity

In the preparation of anti-thrombotic agents the 2- and 3-positions of 3S-tetra-hydroisoquinoline-3-carboxylic acid (THIQA) were simultaneously modified with amino acids to form 20 novel N-(3S-N-aminoacyl-1,2,3,4-tetrahydroisoquinoline-3-carbonyl)amino acids (8a–t). On an in vitro platelet aggregation model 8a–t selectively inhibit ADP-induced platelet aggregation and their IC₅₀ values are leas than 3.5 nM. On an extracorporeal circulation of arterioveinos cannula model of rats both orally and intraveously effective doses of 8a–t are less than 30 nmol/kg. Cerius² based stereoview of explores 8a–t having highly unfolded conformation. 3D QSAR analysis gives the importance of the unfolded conformation to high in vitro anti-platelet aggregation and in vivo anti-thrombotic potency rational understanding.