原核

Objective To construct the prokaryotic expression system for hemagglutinin esterase protein of swine hemagglutinating encephalomyelitis virus 67N strain and provide a candidate gene for the development of specific immunodiagnostic method.

目的构建猪血凝性脑脊髓炎病毒株(67N)血凝素酯酶蛋白原核表达系统，为建立特异性免疫学诊断方法提供备选材料。

It could be useful for the further study of NS1 gene function. Method The NS1 gene of H5N1 subtype AIV was amplified by RT_PCR and cloned to pGEM_T_easy vector. The NS1 gene expressing plasmids was constructed by inserting the target gene fragments into pGEX_4T_1 vectors. The expression proteins were indued by IPTG and analysed by Western blot.

方法采用RT_PCR方法对H5N1亚型AIVNS1基因进行扩增，将PCR产物克隆于pGEM_T_easy载体，将该基因插入pGEX_4T_1中构建NS1基因原核表达载体，转化BL21大肠杆菌后，在IPTG诱导下表达NS1蛋白，Westernblot鉴定表达NS1蛋白。

The CP gene of WVMV-BJ was amplified by RT-PCR, and ligated to the expression vector pET22b.

采用RT-PCR方法自WVMV-BJ的基因组中扩增出其CP基因，连接到原核表达载体pET22b上。

Based on the conclusion of determining the concentration of rChIFN-γprokaryotic expression and titrating antiviral of sf9 cells, to establish the quantitive ELISA for ChIFN-γ.

在测定原核表达rChIFN-γ的浓度和滴定昆虫细胞表达rChIFN-γ的抗病毒的活性的基础上，建立了鸡γ-干扰素的定量ELISA检测方法。

In order to express he Opening Reading Frame fragment of Banana bunchy top virus Hainan isolateDNA2 correctly ,a pair of primers were designed in accordance with the reported BBTV-HN DNA2 gene sequence.

本文根据已报道的香蕉束顶病毒海南分离物(Banana bunchy top virus BBTV-HN)BBTV DNA2的基因序列，设计一对引物，以海南地区表现束顶病症状的香蕉组织总DNA为模板，利用PCR方法克隆了DNA2编码区，将其插入到原核表达载体pGEX-6p-1谷胱苷肽-S-转移酶基因下游。

Rabbit zygotes were cultured in sequence culture system that contains: R1 culture medium which is with low concentration of glucose (1.0 mmol/L) and then R2 culture medium which is with high concentration of glucose (3.5 mmol/L).

试验采用含低浓度葡萄糖(1.0mmol/L)的R1培养液和含高浓度葡萄糖(3.5mmol/L)的R2培养液对兔原核期受精卵进行了体外序贯培养，并与传统的RID单一培养进行比较，研究了兔早期胚胎在体外序贯培养系统中的发育效果。

Prokaryotic expressed protein products of candidate cDNAs were purified by SDS-PAGE and used as immunogens to prepare murine antisera.

5利用SDS-PAGE分离原核表达产物，并以纯化的表达产物为免疫原免疫小鼠，制备抗血清。

The fusion DNA fragments of ag85b-mpb64 and ag85b-mpb64-esat-6 were obtained by PCR andSOE technique. Various DNA vaccines were constructed with the pcDNA3.1: fusion of two genes, and of three genes, bivalent combinations and trivalent combinations(pCA+pCM+pCE6). BALB/c mice were vaccinated with this DNA vaccines.The mice injected withBCG were positive control and the mice injected with pCDNA3.1 and PBS were negative control.The mice were immunized 3 times with 2-wk intervals. The animals in group BCG were only inoculatedsubcutaneously with 1×10~6 CFU BCG at initial vaccination. The serum IgG titers and IgG isotype weredetermined using iELISA coated with M. bovis PPD and rMAE protein expressed and depurated inprokaryotic expression system every week.

同样，利用PCR和SOE技术，获得牛分枝杆菌mpb64-ag85b和mpb64-ag85b-esat-6融合基因，以pCDNA3.1为载体构建了牛分枝杆菌多价组合和多基因融合DNA疫苗：二基因融合(pCDNA3.1-MPB64-Ag85B，简称pCMA)和三基因融合(pCDNA3.1-MPB64-Ag85B-ESAT-6，简称pCMAE)DNA疫苗；二价组合和三价组合(pCA+pCM+pCE6)DNA疫苗，免疫BALB／c小鼠，以牛分枝杆菌BCG免疫组为阳性对照，以pCDNA3.1及PBS免疫组为阴性对照，共免疫3次，每次间隔2周，BCG组仅初免时皮下免疫1次。1免后每周，以原核表达纯化的重组MPB64-Ag85B-ESAT-6蛋白和牛分枝杆菌PPD为包被抗原，以间接ELISA方法检测血清IgG水平及lgG亚类。

Blastocyst rate of embryos co-cultured with pig oviduct epithelium cells came from pig oviduct during diestrus was significantly lower than metestrous. 4. The blastocyst rate of transgenic embryos collected at 24~26 hours and 18~22 hours after hCG-treated reached 45% and 20% respectively.

实验证明在hCG注射后24~26h之间获得的受精卵，在原核显微注射人血清白蛋白基因后体外培养，有45%注射胚发育到囊胚阶段，而在18~22h仅有20%，二者差异显著（P.05）。5。

The Prokarya contains a single kingdom, the BACTERIA, which is divided into two subkingdoms, the ARCHAEA and the EUBACTERIA.

原核生物只包括一个界——细菌界，它又可以被再分为两个亚界：古细菌和真细菌。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失