重组物

A PCR product was amplified using RT-PCR method from total RNA from the abdomens of R strain using the degerated primer. The PCR product was purificated and cloned. The plasmid is pBluescript Ⅱ KS. Recombined plasmid DNA was identified by endoenzyme. The results indicated there were different P450 genes in the PCR product.

运用这对简并引物首次从德国小蠊抗拟除虫菊酯的品系R中扩增出的特异性的DNA片段，经纯化、连接和转化，重组至pBS质粒，筛选阳性克隆，提取重组质粒的DNA，并经初步的酶切鉴定，发现存在着不同的CYP4基因。

Pneumoniae FH strain was cloned and the sequence was analysed by M13 DNA sequencing method. Comparing the PCR product sequcence with MP M-129 strain P1 gene, we found that there are 4 bases different. This may result from the different MP DNA templates. The maximum homology is 98.8%. The result confirmed the fidelity and specificity of the amplified target DNA segment by PCP, and suggested that two categories of MP P1 gene still exist a few differences even in the conservation region. The cloning MP DNA segment was labelled by random hexanucleotide priming, after hybridization, the probe detection was completed using an anti-digoxigenin antibody alkaline phosphatase conjugate, and the substrates 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium. This hybridization system is much superior to the radioactive probe hybridization, because it is safe, easy to handle and has no limitation of decay time. The time required for colormetic detection is also much less than the corresponding autoradiographic exposure time needed to achieve similar detection limits with 32P-labelled probes. The Dig-probes could be used repeatedly, and this made them not only much convenient to use, but also lower the cost, and worthwhile to be used popularly.

将PCR产物进行重组，并将阳性重组质粒，应用M13测序系统对产物进行DNA序列分析，并与MPM-129株P1基因核苷酸进行同源性比较，发现有4个位置的碱基发生了变化，其同源性为98.8％，证实了PCR所扩增DNA片段的准确性和特异性，同时也证实了不同MP组型的P1基因即使在保守区也存在着一定的差异，将克隆的目的DNA片段用异羟基洋地黄毒苷配基用随机引物法标记制备MP DNA探针，杂交后用碱性磷酸酶标记的抗Dig多克隆抗体与杂交体反应，再用BCIP和NBT呈色，制备MP DNA探针，鉴定所扩增片段的特异性，与同位素探针比较，Dig探针不受半衰期限制，可反复使用，而且价格低廉，值得推广使用。

Firstly, the recombinant engineering technology related vaccine preparation was set up, including: 1. the gene knock-out based on linear DNA target molecular in vivo; 2. the selection and counterselection technology of exogenous gene knock in definite site on chromosome; 3. the Gap-repair clone technology in vivo; 4. the recombination technology of single-chain DNA and the overlapping primer; 5. Set up the new selection and counterselection technology of neo-E.

本研究首先建立与疫苗制备相关的重组工程技术包括：①基于线型DNA打靶分子的大肠杆菌体内基因剔除技术，②选择与反选择外源基因定位敲入染色体上特定位点技术，③Gap-repair体内克隆技术，④单链DNA重组技术和重叠引物介导的DNA重组技术，⑤建立&抗生素-酶切位点&选择反选择技术新技术。

Positive rate was relatively high, and the foreign gene could also maintain in testicular tissue for a long time, it could still be detected in the tissue in 7 months after being injected. However. Deferent results could be gained when the detection were made with different primers on different sites of the recombinant plasmid. This could be explained by transgene lost when the foreign gene recombinated and integrated in heterogenetic cell, or transgene degradated by cell nucleases.

然而，我们也看到，在后代检测中，利用不同的引物扩增重组质粒不同部位时，检测结果存在差异，转基因阳性率不同，这种差异说明当外源基因进入到异源细胞内，在重组整合时发生了外源基因丢失的现象，并由此导致外源基因的整合率下降，也可能是受细胞内核酸酶的作用，使得外源基因遭到降解。

Then the fusion protein was primary purified by affinity chromatography. The GST tail of the fusion protein was cleaved by Thrombin. Nucleotide sequence analysis showed that the gdh gene of S.suis possesses the highly conserved motifs typical of family I hexameric GDHs.

猪链球菌2型谷氨酸脱氢酶编码基因克隆、序列分析及原核表达根据已发表的猪链球菌2型的gdh基因设计引物，扩增海安病人分离株Habb中的目的基因，并进行序列测定和分析；构建重组表达载体pGEX4T-2-gdh，在大肠杆菌中表达，并纯化重组蛋白。

Rapid amplification of cDNA end:Retroverse transcription product of total RNA extracted from normal porcine tissue was used as the template,gene specific primers were designed and advantage 2 polymerase mix was used in PCR,of which using porcine genomic DNA as the template:forward primer was designed according to the acquired consensus region of human and pig FGL2 3′ sequences while reverse primer was designed from human FGL2 3′ end downstream sequence;TA cloning.

以猪正常小肠及心脏组织提取新鲜总RNA，反转录后作为模板，设计基因特异性引物，采用Advantage 2 聚合酶混合物进行PCR扩增；依据猪与人FGL2基因3′端已知同源序列设计PCR上游引物，以人FGL2基因3′末端序列设计下游引物，以猪基因组DNA为模板采用Advantage 2 聚合酶混合物进行PCR反应；PCR载体重组质粒DNA亚克隆扩增。

In spite of the low binding affinity between the Mis13/Mis14 dimer and the Mis12/PMF1 dimer,the four proteins form a heterotetramer with a stoichiometry of 1:1:1:1 when these four proteins expressed synchronously,implying that the four subunits can act synergically to form an intact Mis12 complex in vivo.

Mis12复合物在动点上的功能已经较为清楚，但是它们分子间结合方式仍是未知的，为了解析Mis12复合物的分子构架，我们引入了共表达系统在体外表达和重组该复合物。

The biological activity of two immunologically similar recombinant grass carp growth hormone were compared with recombinant tuna Thunnus thynnus growth hormone, as well as an analog of LHRH(D—Ala~6, Pro~9—N—Etylamide—LHRH, LHRH—A).

本文对两种具有相似免疫活性的草鱼基因重组生长激素(r—gGH—Ⅰ、r—gGH—Ⅱ)的生物活性进行了初步研究，并把它们对草鱼鱼种生长的促进能力与金枪鱼基因重组生长激素的一种高效类似物LHRH—A(D—Ala~6，Pro~9—N—Etylamide—LHRH)进行了比较。

After optimizing the expression conditions, a higher production could be obtained when GS115-proCPB was induced in BMGY(pH 6.0), at 28℃, with addition of 0.5% casein acid, 3 mmol/L PMSF, and 0.5% methanol per 12 h. The yield of recombinant protein reached 500 mg/L, and achieved over 94% of total protein in the culture supernatant.

通过表达条件的优化，使用BMGY(pH 6.0)培养基，添加0.5%的酪蛋白水解物和3 mmol/L的PMSF，于28 ℃培养，每隔12 h补加0.5%的甲醇，重组酵母GS115-proCPB 表达的重组蛋白可达500 mg/L 以上，表达的目的蛋白占总蛋白的94%以上。

The packaging systems are based on the expression of helper functions by coinfecting re-combinant poxvirus vectors comprising recombinant polynucleotides.

所述包装系统基于通过包含重组多核苷酸的重组痘病毒载体的共感染表达辅助功能物。

Finally, according to market conditions and market products this article paper analyzes the trends in the development of camera technology, and designs a color night vision camera.

最后根据市场情况和市面上产品的情况分析了摄像机技术的发展趋势，并设计了一款彩色夜视摄像机。

Only person height weeds and the fierce looks stone idles were there.

只有半人深的荒草和龇牙咧嘴的神像。