质粒

Inaddition,we extract total RNA from cultural U251 cell of humanglioma,make coding gene extron amplification of TfR through ReverseTranscription-Polymerase Chain Reaction,construct expressionplasmid of pCDNA3.1-TfR after digestion of enzyme andconjunction,do transfection to U251 cell by liposome afteridentification,cause excess expression,detect the efficiency of transientexpression of TfR protein by pEGFPN1-TfR, sieve positive cell ofpCDNA3.1-TfR through G418, choose positive cell clone, detect theexpression of TfR in anteroposterior period through Western blot,FCM,cell immunochemistry and mRNA through RT-PCR, identify the effect of transfection.

另外，培养人脑胶质瘤U251细胞，进行细胞总RNA抽提，运用RT-PCR方法进行TfR基因的完整编码区外显子的扩增，经过酶切、连接后构建pCDNA3.1-TfR的表达质粒，鉴定正确后按脂质体转染方法转染人胶质瘤U251细胞，使其过量表达，通过pEGFPN1-TfR检测其瞬时转染的效率；用G418对稳定表达pCDNA3.1-TfR的细胞进行筛选，挑选阳性细胞克隆，运用Western blot、流式细胞检测技术和细胞免疫化学以及mRNA半定量的方法检测TfR在转染前后的表达水平，鉴定转染的效果。

The location and structures of sex-pheromone-producing gland in female H.insularis were studied by EAG,GC,SEM,and TEM.These studies showed that thegland situate in the intersegmental membrane between the eighth and ninthabdominal segments,and is an eversible abdominal fold;Many plump cones disturbon the surface of the gland.The glandular cells of 2-day old virgin female H.insularis are arranged in one layer,among which the central cells are columnarepithelial cells and flat on two sides.The nucleus is irregular elliptical.There isevident conjugation between cells and the involution is more in the basal membraneof cell.Microvilli are distributed on the cytoplasmic membrane and linked withendocuticle on which there are many layers of chitin,and the outer cuticule is staineddeeper.The cell contains bubbles,mitochondria,glycogen deposits,roughendoplasmic reticulum and smooth endoplasmic reticulum.

结合触角电位、毛细管气相色谱、扫描电镜、透视电镜等技术对小线角木蠹蛾雌蛾腹尖末端不同组织部位提取物的测定分析以及腺体位置和形态结构的观察发现：小线角木蠹蛾性信息素分泌腺位于腹部末端8～9节之间，是一个由节间膜特化而成的上皮结构，为一可外翻的腹褶，腺体表面分布着饱满的锥形体，羽化后2天未交尾的雌蛾腺体细胞呈单层排列，腹面中央由密集的柱形上皮细胞组成，细胞排列向两侧延伸至背部，其形状由柱形逐渐变为扁平形，细胞核为椭圆形，细胞与细胞间有明显的胞连接，细胞基底膜基褶较多，质膜上分布着微绒毛，并与内表皮连接，内表皮上有多层几丁质，外角质层染色较深，细胞质中含有空泡、线粒体、脂质粒、粗面内质网和光面内质网。

METHODS: BMSCs were obtained from Japanese big-eared rabbits, and in vitro cultured. Then the subcultured BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry.

日本大耳兔骨髓间充质干细胞进行体外培养扩增后，通过pCDNA3.1质粒将碱性成纤维细胞生长因子基因转染至生长状态良好的骨髓间充质干细胞，并将转染后的细胞接种于制备好的猪小肠黏膜下层上，进行体外联合培养，构建组织工程皮肤。

METHODS: Lipofectamine method: The BMMSCs were obtained from the tibias and femurs of the guinea pigs. The third passage BMMSCs were cultured with plasmid-lipofectamine mixture for 6 hours, followed by fetal bovine medium for 48 hours. Immunohistochemistry was performed for transient expression. G418 was added after 48 hours.

①脂质体法：取体外分离、培养的第3代豚鼠骨髓间充质干细胞，将质粒-脂质体混合物加入含细胞的培养基中培养6 h，再加入胎牛血清的培养基，孵育48 h 后行免疫组织化学检测，即为瞬时表达。48 h后加入含G418培养基筛选。

In addition, the stability of plasmids in 11 representative strains during culture was examed.

此外，本研究还考察了11个不同质粒型菌株的质粒在培养条件下的稳定性。

Co-transformation of plasmid pGg-gfp and plasmid pCc1001 which harbors the complementary gene trp1 was conducted by the PEG-mediated protoplast transformation of the oidia of LT2, a tryptophan auxotrophic strain of Coprinus cinereus.

采用PEG介导法把表达质粒pGg-gfp与辅助质粒pCc1001（含有trp1基因）共转化进色氨酸营养缺陷型的灰盖鬼伞粉孢子的原生质体中。

After the animal modle done successfully, all animals were randomly divided into two groups. group l: Microbubble contrast agent containing plasmid was injected into nude mice via the tail vein; group 2: the mixture of microbubbles with plasmid was injected as the group 1, and eyeballs were exposed to ultrasound with intensity of 0.5W/cm^2 immediately, and the time were all 60s that working time control at 20%.

将12只BAlB/C裸小鼠双眼玻璃体腔接种HXO-Rb44细胞，造模成功后，将动物随机分为2组，第1组于尾静脉注入含质粒的微泡造影剂；第2组尾静脉注射质粒与微泡的混合液，并立即以0.5W/平方公分〔1〕的超声波辐照小鼠眼球60s，工作时间控制为20%。

The toxin-antitoxin system was originally found on the low-copy plasmid to maintenance the plasmid stability.

毒素-抗毒素系统最早发现于一些低拷贝的质粒，用来维持低拷贝质粒在菌群中的稳定存在。

The expression of PCDNA〓-EGFP plasmid was monitered withfluorescence microscope,and the human thyroid undifferentiated carconoma in situmodel and haematogenous metastasis model were established in nude mice withPCDNA〓-EGFP positive cells.These model were used in discussing tumourmetastasis mechanism and early diagnosis of secondery tumour.

荧光显微镜观察〓-EGFP质粒表达情况，并用转染了PCDNA〓-EGFP质粒的细胞建立人甲状腺裸鼠原位及血源移植模型，探讨其转移机制及转移灶的早期发现。

A SV40 based shuttle vector pSP189 and African green monkey kidney cell line was appied , which constitute a shuttle vector /mammalian cell system to detect mutagenesis in vitro induced by animal drugs olaqiundox and quinocetone.

本研究将基于SV40病毒的短暂复制型穿梭质粒pSP189与非洲绿猴肾Vero细胞组成穿梭质粒／哺乳动物细胞诱变检测系统，并将该系统应用于兽药喹乙醇和喹烯酮的诱变性检测。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！