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。
Synthesis and industrial production of five hydrogenolysis - debenzylation compounds from hexabenzylhexaazaisowurtzitane
六
芐基六氮雜異伍茲烷五種氫解脫
芐化合物
的合成及工業制備
The barnidipine hydrochloride were prepared via cyclocondensation from m - nitrobenzaldehyde, methyl acetoacetate and anmmonium bicarbonate, hydrolysis, esterification with opticaliy ( s ) - n - benzyl - 3 - py - rrolidinol, recrystallisation and salification
摘要研究了以碳酸氫銨,間硝基苯甲醛和乙酰乙酸甲酯為原料,經縮合、部分水解,再與( s ) - n -
芐基3 -羥基吡咯烷經酯
化、分步重結晶、成鹽得到鹽酸巴尼地平
的工藝路線。
Study on synthesis of benzyl cinnamate in presence of stannic chloride catalysis under microwave irradiation
微波輻射四氯
化錫催
化合成肉桂酸
芐酯
的研究
Synthesis of - benzyl phenylacetonitrile promoted by triphase phase transfer catalyst
芐基苯乙腈
的三相相轉移催
化合成
Study on double carbonylation of benzyl chloride to phenylpyruvic acid by cocl2 1, 10 - phenanthroline catalytic system
鄰菲羅啉催
化氯
芐雙羰
化合成苯丙酮酸
的研究
Poly(vinyl alcohol)has been protected against radiation by benzalation and benzoylation.
聚乙烯醇
的耐輻解可通過
芐叉
化和苯甲酰
化提高。
The cocatalytic effects of k2co3 on the double carbonylation of benzyl chloride
碳酸鉀在吡嗪類鈷配合物對氯
芐雙羰
化反應中
的助催
化作用
Synthesis of high temperature heat - transfer fluid through benzylation of diphenyl
聯苯
芐基
化反應合成高溫導熱油
的研究
The effects of both the ph values of extracts and the grinding of mycelia in liquid nitrogen on the result of dna extracting were also discussed
同時研究比較了液氮研磨和提取液
的ph值對氯
化芐法提取dna效果
的影響。
However, both the ctab and sds - ctab methods were only adequate to be used to extract dna of some strains of penicillium, and the dna bands of some strains were clear, but others were not obvious
另外一些菌株則提取不出譜帶清晰
的dna 。而氯
化芐法適合於供試
的所有青黴菌株
的dna提取,所有菌株
的dna凝膠電泳圖譜帶清晰,效果很好。
Benzyl choeide method, ctab method and sds - ctab method ) were used to extract dna from eleven strains of penicillium, the results showed that the benzyl choeide method was able to be used to extract the dna of those penicillium strains, and bands resulting from gel electrophoretic separations were very clear
比較了ctab 、 sds - ctab 、和氯
化芐這3種dna提取方法提取青黴dna
的效果。實驗結果表明: ctab法和sds - ctab法只能適合於部分青黴菌株
的dna提取。
Benzyl chloride were used for extracting genomic dna of aspergillus. niger 14, about 1. 5kb specific fragment was obtained from genomic dna of aspergillus. niger 14 by pcr amplification with primers ( forward primer5 " ataggcatcatgggcgtctct3 " reverse - primer5 " cagctaagcaaaacactccgc 3 designed according to the known sequences of the phytase gene in the gene bank and pyrobest ? dna polymerase, after ligated with pmd18 - tvector, transformated into e. colidh5a competent cell successfully. 3. nucleotide sequence analysis of the cloned fragment revealed the presence of the whole phya gene in pcr product
用氯
化芐法提取了aspergillus . niger14 ~ #基因組dna ,根據genebank中已知
的黑麴黴植酸酶基因序列設計出一對特異性引物(上游引物: 5 ataggcatcatgggcgtctc3下游引物: 5 cagctaagcaaaacactccgc3 ) ,採用pyrobest ~ ( tm ) dnapolymerase (高保真dna聚合酶) ,通過pcr方法從aspergillus . niger14 ~ #基因組dna中擴增出了預期
的1 . 5kb左右
的特異性產物,將其與pmd18 - tvector連接后,轉
化e . colidh5菌株
的感受態細胞,經質粒抽提、酶切鑒定,確認該目
的產物已得到成功克隆。
Abstract : the application of carbonyl cobalt as a catalyst for carbonylation in organic synthesis is reviewed with 63 references. the carbonylation of benzyl chloride to phenylacetic acid catalyzed by carbonyl cobalt is discussed in great detail
文摘:討論了羰基鈷在有機催
化羰
化反應中
的研究及應用現狀;介紹了羰基鈷催
化合成各種有機羰基
化合物
的方法,重點討論了氯
芐催
化羰
化合成苯乙酸
的研究進展;在此基礎上對羰基鈷在催
化羰
化反應領域
的應用作了展望。
Au. the influence of several variables such as gold and cosolvent concentrations, shaking times, the ratio of tdmba and gold on the extraction of gold and the water concentration of the organic phase were studied. while the cosolvent ratio is more than 10 %, in the tdmba system, gold was extracted completely. the gold extraction mechanism was discussed on the bases of water concentrations of organic phases and infrared spectrum. it was found that the aggregation ratio of tdmba and au
等因素對萃取率
的影響,並研究了有機相
的連續萃取有機相金含量與水含量
的關系。結果表明,助溶劑體積比10 %以上時,十四烷基二甲基
芐基氯
化銨對金具有萃取能力強,平衡時間短
的特點。季銨鹽陽離子與au cn
The factors influencing on the yield of benzyl propionate, such as the dosage of catalyst, molar ratio of benzyl alcohol to propionic acid, reaction time and the amount of cyclohexane and so on, were investigated
考察了催
化劑用量、苯甲醇與丙酸物質
的量比、反應時間和帶水劑環己烷用量等因素對丙酸
芐酯收率
的影響。
The study of catalytic synthesis of butyl acetate under microwave irradiation
微波輻射法催化合成苯甲酸芐酯的研究