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Nucleoprotein is also a major immunogenetic antigen for SARS; however, several tests have shown that N protein had the reactivity with other coronavirus positive sera. It is therefore an advantage of S protein as recombinant target antigen for SARS serum diagnosis compared with recombinant N protein. Moreover, S protein has been shown to be the major antigenic determinant that induces neutralizing antibodies and protective immunity to SARS-CoV, so it is also important in designing SARS genetic engineering vaccine.

SARS-CoV S 蛋白由1255 个氨基酸组成,体外表达如此大的蛋白很困难,而且表达蛋白的可溶性不高;其次,实验已经证实,全长S 蛋白含有大量无关的表位,致使表达的全长S蛋白对SARS 阳性血清的检出率低于S 蛋白片段;另外作为疫苗研究,S 蛋白上大量无关表位作为异体蛋白进入体内,会引发机体的过度免疫反应,这也是SARS-CoV 可能的致病机理之一。

It was then introduced by gene pulser into acrystalliferous strain CryB and a recombinant strain CryB (pBMBZGC10) was obtained. Different fermentative solutions of recombinant strain were used for multi-spray on leaves of Brassica pekinesis, Ipomoea aquatica and Lycopersicon esculentum. Results by using fluorescent detection and PCR amplification revealed that cry1Ac10 gene could not been detected in roots, stems and leaves of plants tested. The cry1Ac10 gene was also not transferred into indigenous bacteria, actinomyces and fungi in soil.

苏云金芽胞杆菌标记重组菌株与杀虫基因水平转移的研究将重组菌株CryB(pBMBZGC10)的发酵液按浓度梯度分次进行喷洒小白菜、蕹菜和番茄作为供试植株,利用标记基因研究的结果表明:cry1Ac10基因没有向供试土壤细菌、真菌和放线菌转移,也未在植物的供试植物根、茎和叶中检测到该基因。

Results We obtained a lysogenic bacteriophage MZTP01with clear plaque and 1 mm diameter. Fragment D with 2362bp (Genebank No. AY639599) was obtained after the phage DNA hydrolyzed by HindⅢ/EcoRⅠ. Among fragment D, the pep gene with molecular weight of 47 kDa and length of 1101bp was cloned and expressed. Recombinant M15 (pQE30pep) was built and overexpressed in Escherichia coli with a 47kDa clear band. At the same place a clear band was observed by Western blot. Judging from the time course expression, we could conclude that PEP protein produced at 1 hour after induction and then increased gradually. PEP protein was mainly in the form of inclusion body in the recombinant and slowed the growth speed of host. Homologous comparison of PEP protein from phage MZTP01 with other PEPs from BLAST were that phage MZTP01 PEP protein had 100% homologe with that of Escherichia coli K12, and most of others took the similarity in the range between 37%~84%.

诱导获得的溶原性噬菌体MZTP01斑点清晰,直径约1mm,成斑时间12h;从噬菌体基因组DNA双酶切(HindⅢ/EcoRⅠ)片段中回收长度为2362bp的D片段(Genbank登录号: AY639599),又从D片段中克隆了长度为1101bp、编码367aa、分子量为47kDa的pep基因,表达载体M15(pQE30pep)在大肠杆菌(Escherichia coli, E.coli)中表达获得了47kDa的清晰表达带,在1h 时开始产生蛋白并有逐步上升的趋势; Western blot 也在47kDa处得到一条清晰的条带;可溶性分析表明PEP蛋白在重组菌株中是以不可溶的包含体形式存在的,该蛋白的产生明显地抑制了宿主的生长速度;噬菌体PEP氨基酸序列之间的同源性比较表明,噬菌体MZTP01 PEP蛋白与来自E.coli K12噬菌体的PEP蛋白的同源性程度最大。

AIM To study the long-term toxicity of modified recombinant human tumor necrosis factor rhTNF-NC in Macaca mulatta compared with recombinant human tumor necrosis factor rhTNF.

目的 研究新型重组人肿瘤坏死因子rhTNF-NC iv对恒河猴的长期毒性,并与重组人肿瘤坏死因子rhTNF作比较。

The Taenia solium oncosphere 18 gene was cloned and reconstructed into expression vector pYA3341.The recombinant plasmid pYA3341-TSOL18 was finally electro-transformed into an attenuated S.typhimurium definitive host strain of X4500 and the recombinant strain of X4500 (pYA3341-TSOL18) was identified its immunogenicity of expression protein, stability, growth curve, safety in vitro and immunity evaluation by animal experiment in mouse.

克隆并改造TSOL18基因,构建重组质粒pYA3341-TSOL18,电转入鼠伤寒沙门氏菌终宿主菌株X4550,体外鉴定重组菌X4550(pYA3341-TSOL18)表达蛋白的免疫原性、稳定性、生长曲线、安全性和小鼠免疫试验进行评价。

The present invention provides important theoretic base for constituting recombinant virus with expanded host domain and provides one hopeful way for developing recombinant virus pesticide with wide spectrum pesticidal activity, and improving and developing genetically the expression vector system.

本发明为构建宿主域拓展的重组病毒提供了重要理论依据,并且为开发出具有广谱杀虫活性的重组病毒杀虫剂以及表达载体系统的遗传改良和开发提供了一条有希望的途径。

Nodosus pilin and the killed D. nodosus vaccines. It also means that the IL-2 in the fused Pilin-IL2 construct may also have a potential role in stimulating vaccine antigen .Key words: D. nodosus Recombinant Pilin gene Recombinant Ovine IL-2 gene Fused Pilin-IL2 gene

用Pilin-IL-2融合基因工程疫苗免疫健康家兔的初步免疫试验结果表明,2l天后产生较高的凝集效价,达到〓,甚至稍高于加佐剂的前三种疫苗,证明绵羊IL-2基因与Pilin基因融合后,可起到一定的免疫佐剂作用。

Results Multicopy recombinant plasmids pDGXHP2.0-2M Fα-HBsAg and pDGXHP2.0-4MFα-HBsAg were constructed. Three recombinant Hansenula polymorpha strains for secretory expression of HBsAg were screened.

结果 构建了HBsAg汉逊酵母多拷贝表达质粒pDGXHP2.0-2MFα-HBsAg和pDGXHP2.0-4MFα-HBsAg,筛选获得了分泌表达HBsAg的汉逊酵母HP/2MFα-HBsAg和HP/4MFα-HBsAg。

The DNA of Yak PrP expressed plasmid extracted from the recombinant expressed bacterium was identified by PCR amplification and two-ezyme digestion. The expressed product was obtained from the recombinant bacterium inducted under the optimized expressed condition (37℃, 1 mmol/L IPTG, 6 h), and it was determined by SDS-PAGE and Western blotting. The GST-BoPrP(23—242) fusion proteins were collected by two ways: the first is to purify and renature from the inclusion bodies; the second is to separate by SDS-PAGE.

将保存的含有牦牛PrP基因重组表达菌提取质粒DNA,进行PCR扩增和双酶切鉴定,并在优化诱导表达条件(37℃,1mmol/L IPTG,6h)下,获得的表达产物进行SDS-PAGE和Western blotting检测;将鉴定之后的重组表达菌诱导表达后,通过两种方法回收GST-BoPrP(23~242)融合蛋白:其一,是从包涵体中提取和复性GST-BoPrP(23~242)融合蛋白;其二,是通过SDS-PAGE直接分离并纯化GST-BoPrP(23~242)融合蛋白。

To express chicken osteoprotegerin in Pichia pastoris and determine its inhibitive effects on mature osteoclastic bone resorptive function in vitro. The chOPG cDNA encoding for 382 amino acid without signal peptice was subcloned into Pichia pastoris expression vector pPICZα-A. The constructed plasmid was transformed into yeast X-33 by electroporation. The recombinant transformants were selected by Zeocin. Induced by the addition of methanol every 24 hours, the product analyzed by SDS-PAGE was sized about 43kDa and 53kDa at a yield of 200mg per litter of culture. The result of Western blotting indicated that the recombinant protein had specific antigenicity mainly owing to heterogeneous glycosylation.

为了在毕赤酵母中分泌表达鸡骨保护素(osteoprotegerin,OPG),采用DNA重组技术将chOPG成熟肽cDNA片段插入毕赤酵母表达载体pPICZα-A中,以电穿孔法转化酵母X-33,用Zeocin 平板筛选重组子,经甲醇诱导表达后, SDS-PAGE 和免疫印迹分析表达产物,由于糖基化不同,表达产物有两种,其相对分子量分别为43kDa和53kDa,表达量约为200mg/L,经Western印迹验证,有较好的抗原性。

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Do not gum up the work by telling mother what we are going to do .

别告诉母亲我们要干的事,那会把事情搞糟的。

In recent years, application of the partial prestressed concrete beam s is developed.

近年来,随着部分预应力砼梁应用的推广,发现我国规范所采用的名义拉应力法的计算结果不稳定。

Gorky once said:"If it is not a book, I'd be stupid and indecent drowning."

高尔基曾说:"如果不是书籍的话,我就要被愚蠢和下流淹死。"