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
Benzyl mercuric chloride
芐基汞
化氯
Using niobic acid as catalyst, cyclohexane as dehydrant, benzyl propionate was synthesized based on propionic acid and benzyl alcohol
摘要以鈮作催
化劑,環己烷為帶水劑,用丙酸和苯甲醇直接合成了丙酸
芐酯。
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
Abstract : using sulfamie acid as catalyst, synthesis of benzyl acetate and phenylethyl acetate through esterification were studied. the yields are good with high purity
文摘:研究了氨基磺酸催
化酯
化合成乙酸
芐酯和乙酸苯乙酯。此工藝反應時間短、能耗低、產品純度高、產率良好、無三廢污染,優于文獻方法。
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
Benzalkonium chloride solution
氯化芐烷銨溶液