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Because the dimeric form is the active form to exert its biological activities, many trials failed to express TGF β s in prokaryotic systems because of their complex folding manner and disulfide patterns. In 1991 Cerletti and coworkers from Ciba-Geigy Corporation in Switzerland successfully finished the functional expression of human TGF β 2 in bacteria, but the details are protected by the patent.

由于双体是其发挥生物活性的形式,TGF β复杂的折叠方式和二硫键配对使其早期原核基因工程研究都以失败而告终。1991年,瑞士Ciba-Geigy公司的Cerletti等成功完成了人TGFβ2基因的细菌表达,但其技术细节被专利加以保护。

The study cloned the flic gene of App and performed sequencing and analysis of biological information. Polymerase chain reaction PCR was used to amplify the flic gene from.

构建猪传染性胸膜肺炎放线杆菌(Actionobacillus pleuropneumoniae, App)鞭毛蛋白编码基因的重组表达质粒,对其编码蛋白进行生物信息学分析,并将其在原核细胞中进行表达。

Producing PHAs,and then were orientationally inserted into a prokaryotic expression vector pBV220. By analyzing the bioactibity and function of their products, the biological function of phaA, phaB and phaC was confirmed. These have been done in our laboratory. To make better use of the three genes by plant bioreactor, two groups of different plant expression vectors that can be used to produce PHAs in different plants were constructed, and the transformation of PHA in Oxalis triangnlaris has been preliminarily reseached.The following is the main results:Three plant expression vectors pBI121-A,pBI121-B and ubquiting-C, which can be used to be transformed into Oxalis triangularis were respectively constructed by using the phaA, phaB and phaC.

菌株的亚克隆基因组片段中,分离出phaA、phaB和phaC三个基因片段,定向克隆至原核表达载体pBV220,通过对其生物活性与功能分析,确证了基因phaA、phaB和phaC的生物学功能;为了通过植物生物反应器对这三个基因进一步研究利用,本实验利用这三个基因构建了两组可用于不同植物生产PHA的真核表达载体,并对紫叶酢浆草进行PHA基因遗传转化的初步研究,获得主要结果如下:利用克隆的phaA、phaB、phaC三个基因分别构建了可用于转化紫叶酢浆草表达载体pBI121-A、pBI121-B、uBquiting-C;可用于玉米转化的种子特异性表达载体p7S-A、pBI121-S-B、uBquiting-C。

Transgenic tobacco plants were obtained through screening with kanamycin. The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants. Pokeweed antiviral protein Ⅱ is expressed with high level in summer leaves. The expression of PAPⅡ is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPⅡ gene by RT-PCR and then the gene was cloned into E. coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPⅡ gene were then transferred into E. coli strain BL21 (DE3)-plysS and Pachia pastoris GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白,本实验以美洲商陆夏季叶片为材料,通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增,对PAPⅡ进行了基因克隆、序列分析,结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9%,然后将该基因构建到原核、真核表达载体上,分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达,在两个表达系统中均获得了有活性的PAPⅡ表达蛋白,体外生物测定表明表达的蛋白均具有抑制病毒的活性,PAPⅡ基因在酵母中还没有表达的报道,这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants.Pokeweed antiviral protein II is expressed with high level in summer leaves. The expression of PAPII is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPII gene by RT-PCR and then the gene was cloned into E.coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPII gene were then transferred into E.coli strain BL21 (DE3)-plysS and Pachia pastor is GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白,本实验以美洲商陆夏季叶片为材料,通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增,对PAPⅡ进行了基因克隆、序列分析,结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9%,然后将该基因构建到原核、真核表达载体上,分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达,在两个表达系统中均获得了有活性的PAPⅡ表达蛋白,体外生物测定表明表达的蛋白均具有抑制病毒的活性,PAPⅡ基因在酵母中还没有表达的报道,这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

Here we summarize the progress of salmon calcitonin's heterogenous expression in prokaryotic and eukaryotic expression systems as well as the amidation of C-terminal. Both the biological activity of salmon calcitonin and the biologic expression system can be improved by genetic engineering techniques.

本文综述了鲑鱼降钙素基因在大肠杆菌、链霉菌、乳酸杆菌和蓝藻等原核细胞,在酵母、昆虫、动植物等真核细胞中异源表达的研究进展,以及通过基因工程的方法能够改进生物表达系统,提高鲑鱼降钙素活性的动态。

Our study designed two pairs of specific primers and cloned two molecular weight of 63ku from Schizaphis graminum, and Rhopalosiphum maize utilizing PCR technique and have done the prokaryotic expression, the main results are as follows:(1)Amplified the Buchnera groEL of Schizaphis graminum, and Rhopalosiphum maize Yangling Biotype at first time all over the world ,and sequenced the full lenghth gene.

本研究设计了特异性引物,利用PCR技术从玉米蚜和麦二叉蚜中克隆出蚜虫Buchnera groEL基因并进行原核表达研究,主要结果如下:(1)从杨凌生物型玉米蚜和麦二叉蚜中在国内外首先扩增出Buchnera groEL基因,测定了全长基因序列,并对相关的同源基因核苷酸进行了比较。

This results, combining pathological, evolutionally biological and genetic evidences, suggested it was possible that foreign transposable elements exist widely both in prokaryotes and in eukaryotes.

并且结合许多病理学、进化生物学及其遗传学的证据对异源转座子在原核和真核生物中广泛存在的可能性进行了讨论。

Methods By the bioinformatics analyzing tools in bioinformatics webs site,a Ta CRISP 2 fulllength gene from the Taenia saginata asiatica fulllength cDNA plasmid libratory was identified and the coding region sequence and the characteristics of the deduced protein were analyzed.The coding region of Ta CRISP was amplified with PCR,endonuclease digestion and cloned into the prokaryotic expression vector pET30a and then expressed in E.coli BL21 with IPTG induction.

用生物信息学方法从亚洲牛带绦虫成虫全长cDNA质粒文库中识别TaCRISP的全长编码基因并预测编码蛋白质的各种结构与功能;将其编码区序列克隆到原核表达载体pET-30a上,测序鉴定重组质粒,之后进行诱导表达,表达产物经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳鉴定。

In most species they are composed of roughly equal amounts of protein and ribosomal RNA , the ribosome consists of two unequally sized rounded subunits arranged on top of each other like a cottage loaf, eukaryotic cells have larger ribosomes than prokaryotic cells but the ribosomes in mitochondria and chloroplasts are about the same size as prokaryotic ribosomes.

在大多数的物种中,蛋白质的量和 rRNA 量是大体相等的。核糖体包括两个大小不同的亚单位,上部结合形成小屋状结构。真核生物细胞中的核糖体要比原核细胞的核糖体大,但是线粒体和叶绿体中的核糖体与原核细胞的核糖体大小相当。

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