养殖褐牙鲆细胞色素b 基因序列的初步研究

对养殖褐牙鲆( Paralichthys olivaceus) 的线粒体DNA Cytb 基因的部分序列进行测定,测得的目的DNA 片段的长 度为410 bp ,其A(104 bp) 、T(119 bp) 、C(117 bp) 、G(70 bp) 4 种碱基平均含量分别为25. 4 %、29. 0 %、28. 5 %、17. 1 %。 在28 个褐牙鲆个体中共出现了3 种单倍型。白化褐牙鲆出现的第1 种和第3 种单倍型个体数分别为10 尾(占白 化褐牙鲆样本数的90. 91 %) 和1 尾(9. 09 %) ;6 尾黑化褐牙鲆均出现第1 种单倍型(100 %) ;正常褐牙鲆出现的3 种 单倍型尾数分别为7 尾(占正常褐牙鲆样本数的55. 56 %) 、2 尾(22. 22 %) 和2 尾(22. 22 %) ;测得的序列与既知序列 间在第6 bp 、第19 bp 和第402 bp 碱基处出现差异。由于褐牙鲆Cytb 基因的高度同源性,研究其白化、黑化和正常 状态时出现的序列差异,对于寻找褐牙鲆白化机理研究的分子标记意义重大。

Study on Mitochondrial DNA Cytochrome b Gene of Chinese sea bass, Lateolabrax sp.

参考鳗鲡等鱼类线粒体 DNA序列进行了中国花鲈线粒体 DNA细胞色素 b基因片断的引物设计、PCR扩增及其序列测定。得到中国花鲈的碱基序列为 4 10 bp,其 A、T、G、C含量分别为 10 1bp(2 4 .6 3% )、112 bp(2 7.32 % )、72 bp(17.56 % )、12 5bp(30 .4 9% ) ,与鳗鲡等其他鱼类相同基因片断序列碱基含量相似。

犬科的线粒体细胞色素b DNA序列及其分子系统学研究

通过对犬科的赤狐、蓝狐、貉和狼4种的线粒体细胞色素b约372bp DNA片段序列分析,结合GenBank中狗、西门豺和非洲野犬3种的该区段DNA序列的比较,共发现113个核苷酸位点存在变异(约30%)。NJ法构建的分子系统树显示,非洲野犬最先从犬科动物中分化出来;犬属的狼、狗和西门豺等3种为系统树上独立的一支,且其分歧的时间较赤狐、蓝狐和貉早;赤狐和蓝狐具有较近的亲缘关系。

中国银额果蝇自然群体B染色体的分布

中国(台湾省外)银额果蝇自然群体中普遍存在Bs。结合前人报道提出,Bs频率的地理分布出现了由东向西和由南向北的规律性的依次升高趋势。这种跨越不同地理环境的区域性梯度变异与其宿主向大陆内地扩散的推论相符。Bs频率最高(87.7%)的是海南岛的孤立隔离群体。大部分群体含Bs的数目多,最多达14条,为标准染色体数目(2n=6)的2.3倍。同时依据该果蝇Bs频率的地理分布特点,将我国现生的银额果蝇划归为5个生态型群体,即沿海丘陵群体、高原型群体、岛屿型群体、山地型群体和峡谷群体等。

Structure and anti-HIV activity of micrandilactones B and C, new nortriterpenoids possessing a unique skeleton from Schisandra micrantha

Two new highly oxygenated nortriterpenoids with a unique norcycloartane skeleton, micrandilactones B and C (1-2), were isolated from Schisandra micrantha; micrandilactone C ( 2) exhibited an EC50 value of 7.71 mu g/mL (SI > 25.94) against HIV-1 replication with minimal cytotoxicity, and the potent anti-HIV-1 activity and unique structural features of 2 make it a promising lead for therapeutic development of a new generation of anti-HIV drug.

肉、奶牛B超活体取卵方法

本发明涉及一种肉、奶牛B超活体取卵方法，属生物技术领域。本方法的吸卵真空泵的压力为50-90mmHg，吸卵液为含0.5-1％,青-链霉素和5-10IU肝素的磷酸缓冲液，洗卵液为含5-12％,BCS的磷酸缓冲液和改良的基础培养液-Hepes液；采卵频率为1次/4天/头；穿刺对象为所有直径≥2mm的卵泡；取1-3级的卵母细胞分装、备用；对7-8岁的供体经产牛按程序注射等量的外源性FSH，总量为30-55μg；对≥14岁的老年牛按程序肌肉注射总量为150-200μg的外源性FSH，以提高卵母细胞发育潜能。本发明的优点在于，从活体可重复、无损伤获取卵母细胞；≥2mm直径卵泡作为穿刺对象和每4天1次的取卵频率保证了卵母细胞的质量和发育同步性；用外源性FSH诱导可提高卵母细胞的发育潜能；本方法的取卵效果佳。

Molecular phylogeny and taxonomy of wood mice (genus Apodemus Kaup, 1829) based on complete mtDNA cytochrome b sequences, with emphasis on Chinese species

Phylogenetic relationships among 15 species of wood mice (genus Apodemus) were reconstructed to explore some long-standing taxonomic problems. The results provided support for the monophyly of the genus Apodemus, but could not reject the hypothesis of paraphyly for this genus. Our data divided the 15 species into four major groups: (1) the Sylvaemus group (A. sylvaticus, A. flavicollis, A. alpicola, and A. uralensis), (2) the Apodemus group (A. peninsulae, A. chevreri, A. agrarius, A. speciosus, A. draco, A. ilex, A. semotus, A. latronum, and A. mystacinus), (3) A. argenteus, and (4) A. gurkha. Our results also suggested that orestes should be a valid subspecies of A. draco rather than an independent species; in contrast, A. ilex from Yunnan may be regarded as a separate species rather than a synonym of orestes or draco. The species level status of A. latronum, tscherga as synonyms of A. uralensis, and A. chevrieri as a valid species and the closest sibling species of A. agrarius were further corroborated by our data. Applying a molecular clock with the divergences of Mus and Rattus set at 12 million years ago (Mya) as a calibration point, it was estimated that five old lineages (A. mystacinus and four major groups above) diverged in the late Miocene (7.82-12.74 Mya). Then the Apodemus group (excluding A. mystacinus) split into two subgroups: agrarius and draco, at about 7.17-9.95 Mya. Four species of the Sylvaemus group were estimated to diverge at about 2.92-5.21 Mya. The Hengduan Mountains Region was hypothesized to have played important roles in Apodemus evolutionary histories since the Pleistocene. (C) 2004 Elsevier Inc. All rights reserved.

与植物多糖硫酸酯结合的HIV-1ⅢB灭活病毒免疫小鼠的研究

目的：为了探讨植物多糖硫酸酯（’&&(）与)*+%$ 结合后，能否诱导,-./+#0 的)*+%$ 暴露出中和抗体的表 位，用它作为灭活疫苗以便诱导产生中和抗体。方法：用’&&( 结合的灭活,-./+#0 作为免疫原，与佐剂混和后，免疫 0(102 3 小鼠，制备出免疫血浆。用41-5( 检测血浆内抗,-./+ 特异性-)6 抗体的滴度，用改良的活细胞染色法中和试验检测 免疫血浆的抗,-./+#0 的中和活性。结果：从与’&&( 结合的,-./+#0 免疫组的动物获得的免疫血浆内抗,-./+ 抗体的滴 度（7 组：+8 # 9 +$" ；: 组：+8 # 9 +$" ）比未结合’&&( 的,-./+#0 免疫组（;8 < 9 +$# ）高，雌性小鼠的免疫血浆的特异性抗体滴 度比雄性的高& 倍。所有免疫组获得的免疫血浆均没有抗,-./+ 中和活性。结论：’&&( 与)*+%$ 相互作用不能诱导暴露出 )*+%$ 的中和抗体表位，但’&&( 可以增强机体免疫原的抗体反应强度，提示它可以作为免疫增强剂用于疫苗研究。

7 种蛇毒小肽对临床分离耐(多) 药结核分枝杆菌的体外活性研究

目的　检测7 种从蛇毒分离的小肽是否对临床分离的耐药性结核分枝杆菌菌株具有活性。方法　放射性方法检测蛇毒 小肽对结核分枝杆菌的最小抑制浓度,细菌存活计数确证放射性方法的结果。结果　7 种蛇毒小肽对耐药性结核分枝杆菌菌 株都有活性。其MIC 值分别为(μg·mL - 1) : Opiophagus hannah 5. 4 , Naja at ra 8. 6 , B ungarus f asciatus 6. 4 , Trimeresurus ste2 jnegri 12. 6 , Protobothrops mucrosquamatus 11. 8 , Protobothrops jerdonii 7 , A gksist rodon halys 4. 2 。结论　这些结果是首次报 道,为进一步设计和开发新来源的抗结核病新药提供了依据。

刺鲃基于线粒体细胞色素b基因的生物地理学过程

以中华倒刺Spinibarbussinensis为外类群 ,研究了不同地理种群刺Spinibarbuscaldwelli细胞色素b基因序列 (114 0bp)变异 ,以探讨其生物地理学过程。结果表明 :长江下游水系与珠江水系种群的变异值为 1 2 %— 2 3% ,与闽江水系的为 2 7%— 3 7% ,与九龙江水系的为 3 1%— 4 2 % ,这些值都远远低于它们与中华倒刺的变异值(13 2 %— 14 6 % )。遗传变异值表明了刺的生物地理学过程 ,首先是东南沿海的水系同内地的水系发生隔离

江汉平原农田生态系统B素循环及平衡分析

以江汉平原农田生态系统为研究对象 ,通过对当地农户小麦 -稻、稻 -稻、油菜 -大豆、油菜 -花生、小麦 -芝麻、小麦 -棉花、青椒 -大白菜、萝卜 -茄子 8种种植模式农田 B素的输入、输出和平衡研究。结果表明 ,B素的输出主要是作物收获 ,占 B素总输出量的 4 4.8%～ 6 4 .7% ;其次是淋溶损失占 2 5 %～ 4 1 .4 % ,B素流失占总输出量的 9.2 %～ 1 7.4 %。B素的主要输入途径是施有机肥和 B肥 ,此外 ,降雨也是 B素输入的主要途径 ,该区域各种类型农田生态系统

Ultraviolet-B exposure induces photo-oxidative damage and subsequent repair strategies in a desert cyanobacterium Microcoleus vaginatus Gom.

Biological soil crusts are important in reversing desertification. Ultraviolet radiation, however, may be detrimental for the development of soil crusts. The cyanobacterium Microcoleus vaginatus can be a dominant species occurring in desert soil crusts all over the world. To investigate the physico-chemical consequences of ultraviolet-B radiation on M. vaginatus, eight parameters including the contents of chlorophyll a, reactive oxygen species, malondialdehyde and proline, as well as the activities of photosynthesis, superoxide dismutase (EC 1.15.1.1), peroxiclase (EC 1.11.1.7) and catalase (EC 1.11.1.6) were determined. As shown by the results of determinations, ultraviolet-B radiation caused decreases both in contents of chlorophyll a and in ratios of variable fluorescence over maximum fluorescence that indicate the growth and photosynthesis of M. vaginatus, besides, increases both in levels of reactive oxygen species and in contents of malondialdehyde and proline, while intensified activities of superoxide dismutase, peroxiclase and catalase reflecting the abilities of enzymatic preventive substances to oxidative stress of the treated cells. Therefore, ultraviolet-B radiation affects the growth of M. vaginatus and leads to oxidative stress in cells. Under ultraviolet-B radiation, the treated cells can improve their antioxidant abilities to alleviate oxidative injury. The change trends of reactive oxygen species, superoxide dismutase, peroxiclase and catalase are synchronous. These results suggest that a balance between the antioxidant system and the reactive oxygen species content may be one part of a complex stress response pathway in which multiple environmental factors including ultraviolet-B radiation affect the Survival of M. vaginatus. (C) 2009 Elsevier Masson SAS. All rights reserved.

Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin

Defensins are a group of cationic antimicrobial peptides which play an important role in the innate immune system by exerting their antimicrobial activity against pathogens. In this study, we cloned a novel beta-defensin cDNA from medaka (Oryzias latipes) by rapid amplification of cDNA ends (RACE) technique. The full-length cDNA consists of 480 bp, and the open reading frame (CRF) of 189 bp encodes a polypeptide of 63 amino acids (aa) with a predicted molecular weight of 7.44 kDa. Its genomic organization was analyzed, and Southern blot detection confirmed that only one copy of beta-defensin exists in the medaka HNI strain. RT-PCR, Western blot and immunohistochemistry detections showed that the beta-defensin transcript and protein could be detected in eyes, liver, kidney, blood, spleen and gill, and obviously prevalent expression was found in eyes. Antimicrobial activity of the medaka beta-defensin was evaluated, and the antibacterial activity-specific to Gram-negative bacteria was revealed. Furthermore, the lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria, was demonstrated to be able to induce about 13-fol up-regulation of the beta-defensin within first 12 h. In addition, promoter and promoter mutagenesis analysis were performed in the medaka beta-defensin. A proximal 100 base pair(bp) sequence (+26 to -73)and the next 1700 bp sequence (-73 to -1755) were demonstrated to be responsible for the basal promoter activity and for the transcription regulation. Three nuclear factor kappa B (NF-kappa B) cis-elements and a Sp1 cis-element were revealed by mutagenesis analysis to exist in the 5' flanking sequence, and they were confirmed to be responsible for the up-regulation of medaka beta-defensin stimulated by LPS. And, the Sp1 cis-element was further revealed to be related to the basal promoter activity, and transcriptional factor II D (TFIID) was found to be in charge of the gene transcription initiation. All the obtained data suggested that the novel medaka beta-defensin should have antimicrobial activity-specific to Gram-negative bacteria, and the antibacterial immune function should be modulated by NF-kappa B and Sp1. (C) 2008 Elsevier Ltd. All rights reserved.

Expressional induction of Paralichthys olivaceus cathepsin B gene in response to virus, poly I:C and lipopolysaccharide

Cathepsin B is a lysosomal cysteine protease of the papain-like enzyme family with multiple biological functions. In this study, Paralichthys olivaceus cathepsin B (PoCatB) cDNA was isolated from flounder embryonic cells (FEC) treated with UV-inactivated grass carp hemorrhage virus (GCHV) and subsequently identified as a vitally induced gene. The full length cDNA of PoCatB is 1801 bp encoding 330-amino acids. The deduced protein has high homology to all known cathepsin B proteins, containing an N-terminal signal peptide, cysteine protease active sites, the occluding loop segment and a glycosylation site, all of which are conserved in the cathepsin B family. PoCatB transcription of FEC cells could be induced by turbot (Scophthalmus maximus) rhabdovirus (SMRV), UV-inactivated SMRV, UV-inactivated GCHV, poly I:C or lipopolysaccharide (LPS), and SMRV or poly I:C was revealed to be most effective among the five inducers. In normal flounder, PoCatB mRNA was detectable in all examined tissues. Moreover, SMRV infection could result in significant upregulation of PoCatB mRNA, predominantly in spleen, head kidney, posterior kidney, intestine, gill and muscle with 18.2,10.9, 24.7,12, 31.5 and 18 fold increases at 72 h post-infection respectively. These results provided the first evidence for the transcriptional induction of cathepsin B in fish by virus and LPS, indicating existence of a novel function in viral defense. (C) 2008 Elsevier Ltd. All rights reserved.

Gene structures and promoter characteristics of interferon regulatory factor 1 (IRF-1), IRF-2 and IRF-7 from snakehead Channa argus

Three interferon regulatory factor (IRF) genes, CaIRF-1, CaIRF-2 and CaIRF-7, and their promoters of snakehead (Channa argus) were cloned and characterized. The CaIRF-1 gene consists of ten exons, spans 4.3 kb and encodes a putative peptide of 299 aa. The CaIRF-2 gene consists of nine exons, spans 8 kb and encodes a putative peptide of 328 aa. The gene organizations of CaIRF-1 and CaIRF-2 are very similar to that of human IRF-1 and IRF-2 except more compact. Comparison of exon-intron organization of the two genes indicated a common evolutionary structure, notably within the exons encoding the DNA binding domain (DBD) of the two factors. The CaIRF-7 gene spans 4.1 kb and encodes a putative peptide of 437 aa. However, the gene organization of CaIRF-7 consisting of ten exons is different to human IRF-7a gene which has an intron in 5' UTR. Three CaIRFs share homology in N-terminal encompassing the DBD that contains a characteristic repeat of tryptophan residues. The promoters of CaIRF-1 and CaIRF-2 genes contain the conserved sites for NF-kappa B and Sp1. The gamma-IFN activation sites (GAS) were found in the promoters of CaIRF-1 and CaIRF-7. The promoter of CaIRF-7 contains conserved interferon stimulating response element (ISRE) which is characteristic of IFN-induced gene promoter, and suggests that there also exist intracellular amplifier circuit in fish IFN signal pathway. Moreover, the element GAAANN oriented in both directions is repeated in CaIRF promoter regions, which confers to further inducibility by IFN. The constitutive expression of CaIRF genes were found to increase obviously in response to induction by the known IFN-inducer poly I:C. (c) 2008 Published by Elsevier Ltd.