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Based on the cloning FeSOD gene of the HZNH, the recombinant prokaryotic expression vector, PQESOa/FeSOD, was constructed and digested with restriction endonuclease BamH I and Pst I to check its construction. The PQE30a/FeSOD was then transformed into E.coli Ml5 and induced with IPTG. The high expression in vitro was obtained and analyzed on SDS-PAGE gel. The*results showed that the target proteins held a 37% portion in whole bacterial proteins and consisted of two parts, the soluble proteins and inclusion bodies. The soluble proteins in the aqueous layer, checked by means of activities of FeSOD enzymes and analyzed by means of activities of isozymogram from PAGE, demonstrated the induced expression proteins had the active nature of FeSOD enzymes.

以克隆的特异种质烟草HZNH的FeSOD基因为基础,构建了原核表达载体PQE30a/FeSOD,经限制性内切酶BamH Ⅰ、Pst Ⅰ双酶切鉴定后,再转化入大肠杆菌M15中,通过IPTG诱导,得到高效体外表达,经SDS-聚丙烯酰胺凝胶电泳检测,表达的目的蛋白占总菌体蛋白的37%,可溶性和包涵体两种形式均有存在,上清中的可溶性蛋白经FeSOD酶活测定和同工酶活性谱带分析,表明诱导表达的上清中的目的蛋白为有活性的FeSOD酶。

In my experimentⅠcompare the purposes of different combinations of endoenzyme(MseⅠ-HindⅢ, MseⅠ-PstⅠ, MseⅠ-EcoRⅠ) and find that MseⅠ-HindⅢeasily finds thepolymorphism by primers combinations suiTab for AFLP analysis.And by means of this there aremany polymorphism strips. So the double endoenzyme of AFLP was exercisedfor the analysis of soybean cytoplasmic male sterility.In the cource of my experiment, 64 primerpairs of MseⅠand HindⅢwere exerted to analyse the twe mtDNA pools which are made ofsterilities and holdings and the polymorphisms of sterilities and holdings.

运用扩增片段长度多态性的方法,对独特的杂交大豆试材进行分析研究,通过比较MseⅠ-HindⅢ、MseⅠ-PstⅠ、MseⅠ-EcoRⅠ不同酶切组合的效果,结果表明MseⅠ-HindⅢ较容易找到多态性引物组合,而且具有较高的多态性,本试验选用MseⅠ-HindⅢ进行双酶切大豆线粒体DNA从而完成AFLP的分析工作,试验用64对MseⅠ和HindⅢ引物对两个不育系及保持系材料的线粒体DNA制成的pool进行AFLP标记分析,分析了不育系与保持系之间的多态性; 4。

In order to obtain symmetry and parallel polarized light,the double Wollaston prism is amended. After getting symmetric polarized light at a big splitting beam angle,an ordinary triangular prism is added so that the symmetric light beam can be divided into parallel light beams.

为了获得对称且平行分束的偏振光,对双Wollaston棱镜进行了修正,在获得大分束角的对称偏振光后,配合普通玻璃三棱镜,使对称分束光变成平行分束光,通过改变三棱镜与双Wollaston棱镜间的距离来改变平行光的剪切距离。

In order to obtain symmetry and parallel polarized light, the double Wollaston prism is amended. After getting symmetric polarized light at a big splitting beam angle, an ordinary triangular prism is added so that the symmetric light beam can be divided into parallel light beams. The distance between the triangular prism and the double Wllaston prism is changed so that the distance of the shearing parallel light can be changed.

为了获得对称且平行分束的偏振光,对双wollaston棱镜进行了修正,在获得大分束角的对称偏振光后,配合普通玻璃三棱镜,使对称分束光变成平行分束光,通过改变三棱镜与双wollaston棱镜间的距离来改变平行光的剪切距离。

The mouse embryonic stem cells cDNA library was screened by yeast two-hybrid assay,some candidate molecules were get,including DDX39 DEAD (Asp-Glu-Ala-Asp box polypeptide 39, BAT1A(HLA-B-associated transcript 1A), NAC1 ( nucleus accumbens-1 ), BicD2 (bicaudal D homolog 2)、MLL3 (myeloid/lymphoid or mixed-lineage leukemia 3). NAC1 and MLL3 were associated with regulation of gene transcription. DDX39 andBAT1A were associated with pre-mRNA processing and mRNA export from nucleus to cytoplasm. BicD2 was associated with the regulation of Golgi body.

通过利用酵母双杂交方法,以小鼠全长Plk1作为诱饵蛋白筛选小鼠胚胎干细胞cDNA文库中与其相互作用的蛋白,获得了DDX39DEAD(Asp-Glu-Ala-Aspbox polypeptide 39、BAT1A(HLA-B-associated transcript 1A)、NAC1(nucleus accumbens-1)、BicD2(bicaudal D homolog 2)、MLL3(myeloid/lymphoid or mixed-lineage leukemia 3)等几个候选蛋白,其中NAC1、MLL3与基因的转录调控有关,DDX39、BAT1A与mRNA前体的剪切、加工及mRNA从细胞核转运到细胞质紧密相关,而BicD2与高尔基体的调控紧密相关,提示我们Plk1与基因转录、mRNA前体的剪切、加工、mRNA的运输及胞质分裂中高尔基体的调控有关。

After the NK gene from recombinant plasmid pUC19-NK was cloned on expression vector pBV220, it was transformed into host cell E. coli HB101. Recombinant plasmid pBV220-NK was extracted for identifying by analysis of restriction enzymes, PCR and sequencing. Studies on recombinant strain's growth and expression showed that heterogene have no distinct effect on host cell. The recombinant plasmid have excellent segregational stability but low structural stability. The results of kinetics of pBV220-NK expression in E.

将重组质粒pUC19-NK上的纳豆激酶基因成功克隆到表达载体pBV220上,并将之转入大肠杆菌HB101中,得到转纳豆激酶基因工程菌,提取重组质粒经单酶切、双酶切及PCR分析验证后,对重组菌的生长及表达进行一系列研究,结果表明:外源纳豆激酶基因的导入对宿主的生长没有明显的影响:重组质粒的稳定性实验表明:该质粒具有良好的分离稳定性,而结构稳定性较差。

Since Watson and Crick have brought forward the model of DNA helix structure and explain about it's reproduce mechanism in molecule level, research work concerning DNA has been continuously lucubrated. However, whether translating inheritance information and rebuilding DNA molecule via menstruating DNA sequence, or to date the most compelling gene project, chromosome protract et. al, there all need a sort of molecule scissors cleaving DNA, DNA cleavage reagent. At present, tool enzyme of cleaving DNA is mostly DNA restriction enzymes founded in 70 years, which only cleave DNA at specific 4-8 base pair sequences that are usually palindromic, so this restricts the at very fast speed developing of gene technology at certain extent.

自从Watson和Crick提出DNA的双螺旋结构并对其复制机制在分子水平上进行解释后,使得对DNA的研究不断深入,但无论是通过测定DNA的序列来破译遗传信息、改造DNA分子,还是目前最令人关注的基因工程、染色体绘制等,都需要一种断裂DNA分子的&剪刀&——DNA断裂试剂,目前断裂DNA的工具酶主要是上个世纪70年代发现的DNA限制性内切酶,但所知的Ⅱ型限制性内切酶的DNA序列识别长度较短(约为4~8个碱基对),而且一般要求回文结构,这就给飞速发展的基因工程技术带来一定的限制。

Two relevant sites of enzymatic digestion were added to the mTNF-α by PCR. The mTNF-α was linked to the 3'end of m/〓 in pGEX4T-1 vector. The prokaryotic expression vector pGEX4T-1m/〓-mTNF-α was constructed successfully. After induction and expression by IPTG, the expression of two kinds of fusion protein is 15% and 12% of total bacteria proteins respectively. The anti-HCC bifunctional antibodies m/〓-mTNF-α were identified by electrophoresis after the inclusion bodies were purified, denature, renature, re-purified, digested by thrombin and further purified.

采用PCR的方法在mTNF-α的两端加上所需要的酶切位点,将之连接在m/〓的3'端,构建原核表达载体pGEX4T-1 m/〓-mTNF-α,通过IPTG的诱导表达之后,两种融合蛋白的表达量分别占细菌总蛋白的15%、12%,表达产物经包涵体的纯化→变性→复性→纯化→凝血酶酶切→进一步纯化后,可以得到纯度为电泳纯的m/〓-mTNF-α抗肝癌双功能抗体。

Thirty-six rats ,weighing(250±10)g, were randomly divided into normal group, experimental group and control group.Orthodontical devices were put between the upper incisor teeth and dens molaris in both experimental group and control group.The corrective force was adjusted to 60g.Salvia miltiorrhiza combination was implanted to each rat in the experimental group every day.The animals of both experimental group and control group were killed at 7,14 and 21 days.The upper dens malarias and periodontal tissue slices were observed under the light microscope and transmission electro microscope,and the X-ray dental films were taken and measured with digitization dental scanning apparatus and its software.

以36只体重(250±10)g的雄性大鼠为样本,随机分为正常组、实验组和对照组,设定不同浓度丹参复合膜,以同体对照方式在正常组进行体内植入实验,在2周内观察不同浓度丹参的牙周组织诱导作用,寻找最佳应用浓度;在实验组和对照组的上切牙与磨牙之间隙安装正畸装置,建立大鼠磨牙移动实验模型,矫治力的大小为60g;实验组每日每只体内植入丹参复合生物膜,两组动物于正畸加力7、14、21d,分三批处死;处死后立即切取双侧上颌磨牙及牙周组织制取标本,制片透射电镜和光镜观察;拍摄X线牙片,用数字化牙科扫描仪及软件对其进行测量分析。

The corrective force was adjusted to 60g.Salvia miltiorrhiza combination was implanted to each rat in the experimental group every day.The animals of both experimental group and control group were killed at 7,14 and 21 days.The upper dens malarias and periodontal tissue slices were observed under the light microscope and transmission electro microscope,and the X-ray dental films were taken and measured with digitization dental scanning apparatus and its software.Result:The most fitting concentration of combined biological membrane was 0.80g;the speed of the remodeling of the periodontal tissue at the compression side in the experimental group was faster than that of the control group.

以36只体重(250±10)g的雄性大鼠为样本,随机分为正常组、实验组和对照组,设定不同浓度丹参复合膜,以同体对照方式在正常组进行体内植入实验,在2周内观察不同浓度丹参的牙周组织诱导作用,寻找最佳应用浓度;在实验组和对照组的上切牙与磨牙之间隙安装正畸装置,建立大鼠磨牙移动实验模型,矫治力的大小为60g;实验组每日每只体内植入丹参复合生物膜,两组动物于正畸加力7、14、21d,分三批处死;处死后立即切取双侧上颌磨牙及牙周组织制取标本,制片透射电镜和光镜观察;拍摄X线牙片,用数字化牙科扫描仪及软件对其进行测量分析。

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