绵羊胞内单精子注射技术

In this study, the possibility of sheep transgenesis by intracytoplasmic sperm injection (ICSI) was assessed. In experiment 1, activation of ovine oocytes matured in vitro in preparation for ICSI has been investigated with 3.42 mmol/L Ca2+ treatment, ionomycin alone and ionomycin followed by 6-dimethylaminopurine (DMAP) after 3-h delay (group 1, 2 and 3, respectively). After activation, the oocytes were then cultured in SOFaaBSA medium. Cleavage rates were significantly (P<0.05) different among three groups (18.4%, 91.8% and 71.7%, respectively). In additional culture, no parthenotes in group 1, whereas 11% and 17.4% in group 2 and 3 developed to the blastocyst stage. Therefore we used the third activation method in the following ICSI tests. In experiment 2, development of ovine oocytes after ICSI was investigated. Thawed semen from two rams was separated by Percoll centrifugation and was used for ICSI or in vitro fertilization (IVF) trails. A total of 71.8% of oocytes reached the 2-cell stage following living sperm injection, which was significantly (p>0.05) different from those following IVF (41.4%) and sham-ICSI (30.2%). After seven days' culture, no sham-injected oocytes developed into the blastocyst stage, although 7% in ICSI and 16.1% in IVF-oocytes developed into the blastocyst stage, but there was no significant difference in ICSI and IVF groups (p>0.05). In the further study, the possibility of sheep transgenesis by ICSI was assessed. After coinjection of ovine oocytes matured in vitro with dead sperm cold to -20 degrees C and exogenous DNA encoding green fluorescent protein (GFP), seventy-three percent of coinjected oocytes developed to 2-cell stage (33/45) and two of them were transgene-expressing embryos. Among ten embryos at the 16-cell stage, all embryonic cells in one transgenic embryo still expressed GFP. Four coinjected blastocysts were thawed and transferred to the uterine of the two progesterone-synchronized recipient ewe. No pregnancies were detected on the 60th day. These results suggested sheep transgenic embryos could be produced by ICSI and further studies should be performed.     

银杏雌雄基因组DNA间的差异性分析

本研究应用RAPD技术,应用300个10bp随机单引物及200对随机双引物组合,检测了雌雄异株银杏基因组DNA的多态性。结果表明:雌雄基因组间具有极高的相似性,在检测到的3450个标记中,仅获得1个与银杏雄性基因组相关的RAPD标记。以该标记为探针,与雌雄银杏基因组DNA的Southern杂交分析,其杂交信号在两性之间表现为限制性片段长度多态性,该结果为寻找银杏早期性别鉴定的探针以及在细胞和分子水平进一步研究其性别问题奠定了基础。     

虎纹捕鸟蛛毒素-Ⅵ的分离纯化与鉴定

通过离子交换及反相HPLC,从虎纹捕鸟蛛毒液中分离得到1种新的哺乳类神经毒素,命名为虎纹捕鸟蛛毒素-Ⅵ（Huwentoxin-Ⅵ,HWTX-Ⅵ）。MALDI-TOF质谱仪分析及氨基酸序列分析表明该多肽毒素分子量为4.44018kDa,序列为H2N-CIGEG VPCDE NDPRC CSGLV VLKKT LHGIW IKSSY CYKCK-COOH,其中6个Cys形成3对二硫键,小鼠脑内注射实验表明,HWTX-Ⅵ对神经系统具有明显的致瘫作用。这种致瘫作用又具有可逆性。     

乳突寄螨雌螨描述(蜱螨亚纲: 革螨股: 寄螨科)

乳突寄螨 Parasitus mammillatus ( Berlese,1 90 4 )雌螨 ,在所查到的文献中 ,仅记载有生殖区、头盖和螯钳的形状 [1 ,2 ] ,无其它描述。国内根据丹东标本曾描述该螨雄螨和后若螨 [3] 。本文根据朝鲜标本详细描述其雌螨。文中测量单位为 μm,括号内为测量均值。图 1- 7　乳突寄螨 Parasitus mammillatus (Berlese)1.背面 ;2 .腹面 ;3.头盖 ;4 .螯钳 ;5 .颚角 ;6 .须肢 ;7.跗节 乳突寄螨 Parasitus mammillatus ( Berlese,1 90 4 )雌螨 (图 1 - 7)体黄色 ,卵圆形 ,长 71 2 - 793( 758) ,宽 4 0 2 - 483( 4 50 )。背板覆盖整个背面 ,前背…     

湖南类石蛛科—新种(蛛形纲:蜘蛛目)

记述了中国湖南省类石蛛科垣蛛属一新种;大围垣蛛Ariadna daweiensis sp.nov。模式标本保存于湖南师范大学生命科学学院,文中量度单位为mm。     

葛藤雌雄蕊发育的研究

通过光学显微镜和扫描电镜对葛藤的雌雄蕊发育进行观察,结果表明：葛藤雌雄蕊的发育及结构基本上符合蝶形花科的胚胎学特征,其花粉粒为2－细胞型,胚囊发育为蓼型。     

北美香柏雌球果的发育

用扫描电镜(SEM)观察了北美香柏 Thuja occidentalis 雌球果的发育过程。在北京,北美香柏的雌球果是在八月初由营养芽转变而来,雌球果一般有4~6对苞片,中间2~3对可育,每一苞片腋部着生两枚胚珠,在可育苞片腋部最先观察到一扁平的隆起,并在其上分化出两个胚珠原基,接着分化出珠被和珠心,最后形成扁平而两侧对称的胚珠。在北美香柏雌球果发育过程中,约一半的雌球果在2~3对可育苞片中位于下面的1~2对的腋部产生3个胚珠原基,中间一个较小,并在以后的发育中逐渐退化。由此推测北美香柏的雌球果可能是由祖先类群中每一苞片具多于2个胚珠的雌球果演化而来。在光镜下对雌球果维管系统的观察发现,传粉前幼小雌球果的苞片内仅有一束维管束,传粉后随着苞片基部的居间生长,有4—8束维管束在苞片内形成,但是新发育的维管束木质部和韧皮部相对位置与正常叶性器官一致,这与在以往报道的柏科植物成熟雌球果的苞片中均有反向维管束的发育不同。北美香柏雌球果早期发育和维管束分析结果支持傅德志和杨亲二提出的解释裸子植物生殖器官形态演化的“苞鳞-种鳞复合体”理论。关键词北美香柏;雌球果的发育;胚珠分化;SEM     

中国豹蛛属一新种(蜘蛛目:狼蛛科)

记述采自我国海南岛的豹蛛属Pardosa 1新种,针豹蛛P.aciculifera sp.nov.,新种以其独特的外雌器和触肢器而易与本属其它已知种区别。由于其外雌器和触肢器十分独特,目前暂不能将新种归入本属已划定的种组,模式标本存于中国科学院动物研究所动物标本馆。