fmdv 中文意思是什麼

In this paper, first strand cdna of 3abc gene was synthesized using template rna extracted from cells infected with fmdv. the complete 3abc gene about isoobp was amplified by pcr and ligated into pgem - t easy vector. after transforming e. coli dh5 a, ampicillin resistant colonies were isolated and plasmid dna was prepared and analyzed by restriction analysis and pcr. presence of the full length 3abc gene was verified by nucleotide sequence analysis and the plasmid containing the expected sequence was named as pgem - 3abc. comparing the aquired sequence of 3abc with that of reference strains, the homology is more than 99 percent. the pgem - 3abc was digested with sal i and bgl ii and ligated into xho i and bgl ii - digested expression vector ptriex - 4 neo. lt was identified by restriction analysis and pcr and sequencing that this fragment had a 17bp deletion hi the nucleotide sequence 708bp of 3abc gene, which happened to form a terminator codon behind 3ab gene, but it contained the complete open reading frame ( orf ) of 3ab gene. positive clones were selected and induced with lmmol / l isopropyl - d - galactoside ( iptg ), bacteria were detected by sds - page and western blotting after properly treated. the results showed that the 3ab gene expressed successfully in e. coli and 33. 5ku fusion protein can be recognized by the positive bovine serum of fmdv. the amount of target protein is over 26 % of the total bacteria protein by gel thin layer scanning analysis

擴增產物連接到pgem - teasy載體中，轉化大腸桿菌dh5菌株，篩選氨芐青霉素抗性菌落，提取質粒經酶切鑒定、 pcr分析以及確證性測序證明，所克隆的1500bp左右的片段含有完整的3abc基因，與國外參考序列相比，同源性在99以上。將重組質粒pgem - 3abc和表達載體ptriex - 4neo分別用sal和bgl與xho和bgl消化后，亞克隆3abc基因至原核表達載體ptriex - 4neo中，通過酶切鑒定、 pcr擴增以及序列分析，發現克隆到ptriex - 4neo載體上的片段於3abc基因708bp處出現了17bp的缺失，碰巧在3ab基因后形成一終止密碼子，但3ab基因的閱讀框架完整，選出含有3ab基因完整閱讀框架的陽性克隆，用iptg誘導表達，收集菌液進行sds - page電泳、 westernblotting分析，結果表明， 3ab基因在大腸桿菌中成功表達，其表達產物為分子量33 . 5ku的融合蛋白，並能被口蹄疫病毒陽性血清識別。經薄層掃描分析，表達量占總蛋白量的26以上。

Foot - and - mouth disease virus ( fmdv ) is the etiological agent of an important disease of livestock

口蹄疫是當今世界危害最嚴重的家畜傳染病之一。

In the third part of the thesis, a chlamydomonas reinhardtii chloroplast expression vector, pactbvpl, containing the fusion of the foot and mouth disease virus ( fmdv ) vp1 gene and the cholera toxin b subunit ( ctb ) gene was constructed. transformation of c. reinhardtii chloroplast was achieved by biolistic bombardment with pactbvpl

論文第三部分主要敘述了將o型fmdvvp1與強黏膜免疫佐劑霍亂毒素b亞基（ ctb ）的融合基因克隆重組到衣藻葉綠體表達載體中，並採用基因槍法轉化衣藻葉綠體，獲得了具有壯觀黴素抗性的轉化子。

First, a pair of pcr primers was designed to isolate fmdv - vp1 gene according to the published fmdv - vp1 sequence. after pcr of dna isolated from the tissues, the fmdv - vp1 gene was cloned into cloning vector. positive clones were analysed with restriction enzyme digestions and further identified with sequence analysis

首先，根據已知的fmdv基因序列，設計特異性引物，擴增出fmdv - vp1基因，將克隆到的vp1基因連接到測序載體上後用dna測序儀對該序列進行測序。

On our conjecture, gene 3a, a highly conserved sequence, may be the determinant of the virulence of fmdv. changes in 3a have been associated with altered host range. a deletion in 3a has been associated with bovine attenuation of fmdv

我們根據已有資料推測， 3a基因可能是口蹄疫病毒的毒力決定簇，它具有高度保守性，其變異和部分缺失對病毒在牛體內的致弱和宿主嗜性發生改變起重要作用。

Second, a prokaryotic expression construct, obtained from invitrogen transformed into prokaryotic and induced to express vp1 protein. the expressed vp1 fusion protein was purified by affinity chromatography using glutathione - agarose resin and used in elisa and western bolt analysis as the antigen. the elisa and western blot results showed that the anti - fmdv antibody was elicited specifically against vp1 antigen

第二，為了得到抗原蛋白，將vp1的原核表達質粒pgex - 4t - vp1轉化入大腸桿菌bl21中，經iptg誘導，裂解細胞後用瓊脂糖珠進行純化，用elisa和westernblot進行檢測，結果表明誘導表達出所需大小的融合蛋白。

The structural protein vp1 of fmdv carries critical epitopes which can induce neutralizing antibodies

口蹄疫病毒的主要抗原組分為vp1結構蛋白，它能夠誘導機體產生保護性的抗體反應。

In the second part of the thesis, we described that a tobacco chloroplast expression vector, ptrvp1, containing the foot and mouth disease virus ( fmdv ) vp1 gene and the selective marker aada gene was constructed and transfered to the tobacci chloroplast genome by the biolistic method

論文第二部分主要敘述了煙草葉綠體表達載體ptrvp1的構建，並通過基因槍方法轉化煙草葉綠體基因組，獲得了3株具有壯觀黴素抗性的轉化再生植株。

To determine the site - specific integration of the foreign gene into the chloroplast genome and the level of homogeneity, the three transformation lines of the second selective generation were analysized through pcr ( pcr - southern blot ) analysis using the primers of fmdv vp1 gene and homologous fragment of tobacco chloroplast, respectively

經過兩輪抗生素篩選后，採用fmdvvp1基因的一對引物和煙草葉綠體同源片段的一對引物分別對抗性植株的葉綠體dna進行的pcr和pcr - southernblot分析。

Western blot and elisa assays indicated that fmdv vp1 gene was expressed in tobacco chloroplasts and accounted for 2 % ~ 3 % of the total soluble protein ( about 20 - 30 ( ig of chloroplast - expressed vp1 protein in 1 mg of total soluble protein )

Westernblot和elisa免疫雜交分析證明fmdvvp1抗原蛋白在煙草葉綠體中得到表達，表達量占可溶性總蛋白量的2 3 （ 1mg可溶性總蛋白含有20 30 g的vp1蛋白） 。

In this study, pichia pastoris system had been utilized for expression of fmdv 2c3abc gene which aimed for establishing a sensitive and specific molecular dignosis method. first, 2c and 3abc genes were amplified individually from p2 and 3abc postive clones and ligated together using pcr method, then this 2c3abc product was cloned into pgem - t easy vector and transformed e. coli dh5a competent cell. a postive recombinant plasmid which contained whole 2c3abc gene had been confirmed by pcr, enzyme digestion and sequencing. after that, the 2c3abc gene was sub - cloned into ppiczaa expression vector and transformed e. coli dh5 a competent cell and selected by zeocin ? antibiotic. the postive recombinant expression vector was linerized and electro - transformed pichia pastoris smd1168 competent cell. a recombinant pichia pastoris had been obtained by zeocin ? antibiotics selection and induced with 0. 5 % methanol for target protein expression. the expression product was analysised by sds - page and western blotting assay. the result sh owed that 2c3abc gene was expressed successfully in pichia pastoris and the product was a 95ku fusion protein which could be recognized by anti - fmdv serum. the amount of target protein was over 15 % of the total bacteria protein by gel thin layer scanning analysis. this research had supplied materials for establishing a fmd diagnosis method to differentiate infected animals from vaccinated animals

首先，用p2和3abc陽性克隆通過連接pcr方法獲得目的基因並將其克隆到pgem - teasy載體上，並轉化e colidh5a感受態細胞中，經pcr 、酶切以及測序證明得到了完整的2c3abc基因，並與國內外參考序列進行比較分析。然後，將目的基因亞克隆于ppiczaa表達載體並轉化大腸桿菌dh5a ，以zeocin ~ （ tm ）抗性篩選陽性克隆，大量提取重組表達質粒並用pme酶線性化后電轉化入畢赤酵母smd1168感受態細胞，通過zeocin ~ （ tm ）抗生素梯度濃度篩選，獲得重組酵母用0 . 5甲醇誘導表達，通過sds - page電泳、 westernblotting分析，結果表明， 2c3abc基因在畢赤酵母中成功表達，其表達產物為一95ku的融合蛋白，並能被口蹄疫病毒陽性血清識別。

Cloning and prokaryotic expression of 3ab gene of fmdv

基因的克隆與原核表達

As a result, developing a safe and effective fmdv vaccine has become a research priority

目前研究人員通過基因工程方法開發口蹄疫安全的新型疫苗成為研究的熱點。

The sequencing results show that the fmdv - vp1 has mutated compared with the published sequences. and at the same time the 3 dimension protein structure was built for research

通過與genebank中所有的vp1序列進行比較我們發現vp1第24 、 33 、 204位氨基酸突變幅度最大。

Moreover, mice feeding on fmdv - vp1 transgenic plant leaves in their diet produced a virus - specific immune response that could protect the immunized mice against fmdv challenge

另外利用轉基因植物表達口蹄疫病毒vp1抗原蛋白具有安全廉價、蛋白折疊充分、免疫原性良好等特點。

This study is to express fmdv - vp1 protein using the expression system and analysize the protein structure. in this study bioinformatic method is also used to research the mutation and epidemiology principles of the fmdv

口蹄疫是口蹄疫病毒（ footandmouthdiseasevirus ， fmdv ）感染引起的偶蹄動物的急性、熱性、高度接觸性傳染病。

The use of transgenic plants for producing fmdv vp1 antigen protein has been reported and may prove to be safer than conventional vaccines. recently, fmdv vp1 protein had be expressed in transgenic plants

最近利用轉基因植物表達的口蹄疫病毒vp1蛋白注射和飼喂小鼠后，能夠誘導其體內產生特異性中和抗體，並使小鼠得到有效的免疫保護。

Third, the cloned fmdv - vp1 gene was subcloned into an eukaryotic expression vector, psupery - vp1. to evaluate the immune responses of this construct, animals sera were used to analyzed their specific antibodies against vp1 using western blot analysis

第四， dna疫苗因其制備簡單、使用和保存便利，能構誘導機體產生體液和細胞免疫等優點，成為新一代疫苗研究的熱點。