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。

肉牛B超活体取卵效果的研究

对肉牛(莫累灰×婆罗门) 利用B 超活体取卵,研究不同年龄和促卵泡素(FSH) 对平均卵泡数、平均卵母细 胞数、平均可利用卵母细胞(COC) 数、卵母细胞回收率、COC 回收率、囊胚率和平均囊胚数的影响。结果表明:肉牛 B 超活体所取卵的数量和质量受年龄的影响,青年牛好于经产牛,经产牛好于老年牛;适量注射FSH 可以提高经产 牛和老年牛的囊胚率和(或) 平均囊胚数。

从线粒体Cyt b基因探讨红瘰疣螈物种地位的有效性

疣螈属的红瘰疣螈(Tylototriton shanjing)和棕黑疣螈(T.verrucosus)的物种界限一直不清楚.测定了来自中国西南地区14个地点的T.shanjing和T.verrucosus共40只标本的线粒体DNA Cyt b基因(753 bp).结果表明:(1)用邻接法、最大简约法和贝叶斯法等3种系统发育分析法分别重建棕黑疣螈种组系统发育树的拓扑结构不支持T.shanjing是单系群;(2)T.shanjing与T.verrucsus的mtDNA Cyt b序列差异平均值仅为1.2%.未达到种级水平.因此,全部T.shanjing样品都属于同一个物种,即T.verrucosus,不支持T.shanjing的物种地位,T.shanjing为T.verrucosus的同物异名,并建议恢复T.verrucosus的中文名红瘰疣螈.根据基于40个样品Cvt b基因序列的系统发育树和遗传变异以及地理分布,这些红瘰疣螈(T.verrucosus)样品聚为3支,即中国西南地区的红瘰疣螈可分为片马、滇中滇西和滇东南3个地理居群.

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.

HIV-1感染对T和B细胞核内UNG2 mRNA表达的影响

目的:探讨HIV-1感染是否影响细胞中UNG2的表达.方法:采用四步法SYBR green Ⅰ实时定量RT-PCR,对HIV-1感染者的T和B淋巴细胞,以及HIV-1感染的C8166细胞核内UNG2 mRNA的表达进行测定.结果:UNG2 mRNA的表达在HIV-1感染者的T细胞和HIV-1感染的C8166细胞中被明显上调,分别是对照的8.76倍和8.14倍,而在HIV-1感染者的B细胞中却没有被上调.结论:HIV-1感染导致的UNG2表达上调,可能通过减少TCR的多样性削弱Th的功能,另一方面可能有利于病毒对UNG2的包装.

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。

银额果蝇的B染色体研究 1: 昆明群体的Bs数目和频率

银额果蝇昆明群体有丝分裂中期核型中存在B染色体, 出现频率为69.1%在已 研究过的来自各个地区的银额果蝇中, 昆明群体的B染色体频率最高。B染色体 数目为1—6条。该群体内单雌系间的B染色体数目不同, 个体间和细胞间的B染色 体数目也不同。在核型中, B染色体最小, 形态稳定, 点状, C-带和G-带呈阳性 。 图版1图2表1参12

银额果蝇的B染色体研究2: 昆明群体的生活力和B染色体的关系

银额果蝇的B染色体在其生长和发育过程中具双重性调节作用。一方面, B染色体以单一的形式存在于单雌系核型中, 即1B或2Bs可刺激生长发育,增强生活力。另一方面, 在单雌系的细胞中存在多条B染色体, 并形成核型多态性, 似乎对生长发育有一定的抑制作用而减弱其生活力。图1表1参5

Stability and molecular modeling of triplex DNA inhibiting DNA binding protein binding to the core promoter of hepatitis B virus.

Two three-dimensional structure models of the 21nt oligodeoxyribonucleotides, CPI (G3TG-2TGT2G5TG2TGT) and CP3 (TGTG2TGST2GTG2TG3), were constructed by InsightII (MSI) software in IRIS Indigo2 (SGI) workstation using the crystal structure of TAT tripler formation as the template. The initial structures subsequently were minimized by molecular mechanics. The final structures were believed as the dominant conformation. The results showed that the energy of CP1 is lower than that of CP3, and the former is more stable than the latter. Moreover, the results further proved that the 21nt oligodeoxyribo-nucleotide CP1 stably combines with the core promoter (Cp) fragment of hepatitis B virus (HBV) to form a tripler DNA, and CP1 specifically inhibits a specific cellular factor (DNA binding protein) binding to Cp fragment. These results indicated that specific repression of gene transcription of HBV DNA might be possible by tripler-formation DNA.

Molecular phylogeny and biogeography of woolly flying squirrel (Rodentia : Sciuridae), inferred from mitochondrial cytochrome b gene sequences

To investigate the genetic diversity between the populations of woolly flying squirrels (Eupetaurus) from the eastern and western extremes of the Himalayas, partial mitochondrial cytochrome b gene sequences (390-810bp) that were determined from the museum specimens were analyzed using maximum parsimony (MP) and maximum likelihood (ML) methods. The molecular data reveal that the two specimens that were collected in northwestern Yunnan (China) are members of the genus Eupetaurus. Reconstructed phylogenetic relationships show that the populations of Eupetaurus in the eastern and western extremes of the Himalayas are two distinct species with significant genetic differences (12%) and diverged about 10.8 million years ago. Eupetaurus is significantly different from Petaurista and Pteromys. The level of estimated pairwise-sequence divergence observed between Eupetaurus and Petaurista or Pteromys is greater than that observed between Eupetaurus and Trogopterus, Belomys, Glaucomys, or Hylopetes. Considering the divergence time of the two Eupetaurus groups, the glaciations and the uplift of the Himalayas and Qinghai-Tibet plateau during the Pliocene-Pleistocene period might be the major factors affecting the present distribution of Eupetaurus along the Himalayas. (C) 2004 Elsevier Inc. All rights reserved.

增强UV-B辐射下一氧化氮对小球藻氮素代谢和类囊体的保护作用

由于到达地球表面的紫外线B辐射不断加强,生物生长受到了威胁。UV-B的增强改变了生物体赖以生存的环境,影响了藻类生物生长,抑制了其光合作用。以BG11为培养基,在室内培养的条件是光照强度为60μmol·m-2s-(1昼夜比为12h∶12h),温度为26℃,研究了一氧化氮(NO)在增强UV-B(强度为0.2J·m-2s-1)辐射下的对小球藻的作用。测定了小球藻的硝酸还原酶(NR,nitrate reductase)、亚硝酸还原酶(NiR,nitrite reductase)、谷胱甘肽还原酶(GS,gluta

天然三倍体鲫鱼突变体萍乡肉红鲫线粒体细胞色素b基因序列分析

用细胞色素b(Cyt b)基因特异性引物,对萍乡肉红鲫(Carassius auratus var.pingxiangnensis)的线粒体Cytb基因进行PCR扩增和双向测序。在12个个体中均得到序列一致的Cytb基因全序列,长度为1 140 bp。其A、T、G、C含量分别为28.2%(321)、28.8%(328)、14.8%(170)和28.2%(321),A+T含量(57%)明显高于G+C含量(43%),与其他水生动物相同基因片段碱基含量相似。该基因中密码子第1位核苷酸中4种碱基组成较为均衡;第2