受精卵

Methods: Genes of MCP and DAF were connected by 10aa coding sequence of enriching Gly and Ser. Then insert into pcDAN3.1 vector, constituting pcDNA-MD3 expression plasmid of hMCP-DAF fusion gene. The genes were transformed by liposome into the pig endothelial cell; human serum handles masculine cloning, identify the function that restrain breaking of person alexin.

将MCP、DAF两个基因以富含Gly和Ser的10aa编码序列作铰链串接，并接入表达载体pcDNA 3.1中，构成含hMCP-DAF的真核表达质粒(pcDNA-MD3)；以脂质体转染猪内皮细胞，人血清处理阳性克隆细胞，鉴定抑制人补体溶破的功能；应用显微注射将线性基因导入猪受精卵；PCR检测导入基因整合率。

Animals develop from a single cell, a fertilised egg, which divides to produce cells that will form the embryo.

动物们从一颗受精卵衍化而来，受精卵产生组成胚胎的细胞。

Results: in this study,2840 zygotes were injected,the survived 2256 zygotes after microinjection were reimplanted into the oviducts of 85 psudopregnant km female mice.

结果： 注射受精卵2840枚，筛选注射后成活的受精卵共2256枚，注射成活率79.4％。

RESULTS:①A total of 178 injected and survived zygotes were replanted. Thirteen mice were born with the birth rate of 7%.

结果：①178只注射pcDNA3.1-mutant LMNA后的存活受精卵移植至假孕鼠输卵管后，出生13只子代鼠，出生率只有7%；58只注射pcDNA31 -mutant LMNA后的存活受精卵移植到假孕鼠输卵管后，在移植后12~15 d的受体母鼠子宫内收集到1只活胚和5只死胚。

There was no different in culture between different timing of micromanipulation. Zygotes with good quality grew better. Zygotes with bad quality almost cannot grow.

结果：基因浓度越高，受精卵发育越差，操作时间不同的卵发育无明显差异，质量好的受精卵发育较好，质量差的几乎不发育。

When the other conditions were stable, the effect of different concentration of the microinjected DNA, and different timing of micromanipulation in CZB medium, different quality of zygotes on microinjection was investigated in vitro. Results: The higher the concentration of the microinjected DNA, the worse the development of zygotes.

获取小鼠受精卵，在其他条件固定的情况下，分别比较不同的注射基因浓度，在CZB操作液中显微操作时间以及注射前受精卵的质量对显微注射后的胚胎体外培养结果的影响。

At 24 h, Ang1 was expressed mainly in the lens, optic cup, mesencephalon, ventral part of hindbrain, and myotomes in the somites, while zfTie2 was mainly expressed in intersomitic vessels including dorsal aorta and axial veins, and sprouting vessels from the dorsal aorta in the trunk. VEGF was expressed mainly in the lens, optic cup, mesencephalon and myotomes in the somites, while zfFlk1 was mainly expressed in intersomitic vessels including dorsal aorta and axial veins, and sprouting vessels from the dorsal aorta in the trunk.

结果从受精卵胚胎发育第12小时可见血管内皮细胞生长因子和血管发生素1及其相应受体flk1和Tie2的表达；在受精卵胚胎发育的第16小时，血管内皮细胞生长因子和血管发生素1主要在头部和尾部胚芽表达；第24小时在眼球、视窝、中脑、后脑的腹前侧和肌节中表达，Tie2和Flk1主要在肌间背部动脉血管、躯干新生血管芽和轴向静脉中表达。

Oocyte samples from one group were collected to detect the presence and integration of HBV DNA within cells and chromosomes using PCR, Southern blot, dot hybridization and fluorescence in situ hybridization. The female animals from another group were mated with their normal males, respectively. Their zygotes, 2-cell embryos were collected to detect the integration of HBV DNA in the female pronuclei of zygotes and the replication and expression of HBV genes in the 2-cell embryos using FISH, RT-PCR and immunofluoresence assay.(1) PCR detected positive bands in the tested oocyte samples fromgoldon hamster and mice. Southern blot revealed clear hybridization signals in PCR products.

研究用金黄地鼠和小鼠建立实验动物模型：将卵巢内注射HBV DNA的实验动物分成两组，一组注射后进行超排卵，收集卵巢和输卵管的卵母细胞，用PCR、Southern杂交，斑点杂交和荧光原位杂交(fluorescence in situ hybridization、FISH)检测HBV在卵母细胞内的存在和染色体上的整合；另一组超排雌鼠与正常雄鼠合笼，收集受精卵和2-细胞胚，用FISH、RT-PCR和免疫荧光检测技术分别研究HBV基因在受精卵雌原核上的整合以及在2-细胞胚中的复制与表达。

Assays of AKT and MPF activities AKT or MPF kinase activity was measured by histone H2B kinase or histone H1 kinase assay.The frozen eggs were treated with the same method as we previously reported.The radioactivity on the filter paper was counted with a BECKMAN scintillation counter.

免疫荧光分析的结果显示，结合Ser473位磷酸化AKT的绿色荧光FITC在GV期卵母细胞中均匀分布在胞浆中，而在S期的受精卵中绿色荧光分布于细胞膜，AKT的膜定位表明在S期的受精卵中它可能处于活化状态。

The common scenario is that a certain percentage of eggs are bound to be unfertilised....and if they are laid very close to each other, fungus that grow on unfertilised and dead cell will also encase and suffocate the rest of the fertilised eggs and growing fetus.

通常一定比例的卵肯定无法受精…而且如果卵堆挤在一起，那生长在未受精卵和死亡细胞的霉菌会包住并闷死受精卵，长出新的霉菌。

Plunder melds and run with this jewel!

掠夺melds和运行与此宝石！

My dream is to be a crazy growing tree and extend at the edge between the city and the forest.

此刻，也许正是在通往天国的路上，我体验着这白色的晕旋。