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The purified N protein was used to coat 96-well plate, each step was optimized, such as coating concentration of recombinant nucleocapsid protein, scample dilutim, chromogen and stop solution as well as concentration.

在此基础上,以纯化的重组N蛋白作为包被抗原,对各种条件进行优化(如抗原的包被,作用时间及底物的选择),确定了判定标准,建立了检测PRRS抗体的间接ELISA方法。

After washed, the chromogen substrate was added and incubated for 30 minutes in the dark. Rinsed with distilled water, purple spots were observed and counted using a dissection microscope. The number of spots means the level of cytokine producted by sCTL. Analysis of ability of specific cytotoxic lymphocytes to lyse target cells: CytoTox96 Non-Radioactive Cytotoxicity Assay kit was used.

5特异性CTL裂解靶细胞能力测定:采用CytoTox96 Non-RadioactiveCytotoxicity Assay试剂盒,将经HBV core18-27短肽、重组乙肝病毒核心抗原共刺激培养13日的PBMC与HLA-A2〓的靶细胞按一定比例混合,靶细胞被裂解后,释放的乳酸脱氢酶与底物反应显色,酶标仪测定吸光度,吸光度与裂解程度成正比,按照公式计算百分裂解率。

Methods: Two sets of primers were designed according to the cyclostyle of pcDNA3.1-HBV-YVDD, and the full-length of HBV genome with different enzyme digestin spot was amplified.

以pcDNA3.1-HBV-YVDD为模板设计两对引物,扩增带不同酶切位点的HBV全基因,扩增产物纯化后,经酶切、连接,得到pcDNA3.1-HBV-YVDD重组体。

As to the plenty of include bodies in the precipitation , denationalization, detergence, purification and dissolution, last regeneration were recommended to acquire great deal of expressed GST fusion proteins.

COlt BLZI生产融合蛋白,重组后的 DNA序列包括一个PGEX质粒,依照PGEX质粒的融合蛋白的表达是在tao启动子的控制之下,而枯启动子可由乳糖的类似物I刚来诱导它表达。

III For constructing the expression vector of a fusion protein and obtain a target protein with full identity on aa sequence of a natural 13- 1,3-1 ,4-glucanase, with the recombined plasmid DNA harbouring the target gene as template, the primers designed with restriction sites for both terminals and enterokinase recognition site, followed by PCR amplification, was induced to the target gene.

为构建融合蛋白表达载体和获得与天然蛋白质序列完全一致的目的蛋白,以含有目的基因的重组质粒DNA为模板,设计引物时加入两端酶切位点及肠激酶裂解位点,通过PCR扩增引入目的基因中,测序结果表明接头和读框正确。

AIM: To obtain a high and stable expression analog of human basic fibroblast growth factor by genetic engineering.

目的:用基因工程的方法获得高效稳定表达的重组人碱性成纤维细胞生长因子结构类似物。

The flic gene of App was obtained by PCR based on highly homology with Escherichia coli, Salmonella, Shigellosis. Then it was legated to pMD-18T vector. The recombinant plasmid was identified by enzyme and sequence analysis. The 528 bp fragment of flic gene was successfully cloned into pGEX-KG vector, then the recombinant plasmid was transformed into E.

以猪传染性胸膜肺炎放线杆菌血清Ⅰ型菌株基因组为模板,根据其鞭毛区与其它细菌高度同源性设计引物,利用PCR方法扩增鞭毛蛋白基因flic片段,将其亚克隆到pMD-18T载体中并进行PCR和酶切鉴定,再将其亚克隆与载体pGEx-kG分别双酶切和连接,构建重组表达载体pGEX-flic,并将其转化大肠杆菌工程菌BL21,进行原核表达。

Genes important for male gametogenesis involved in highly con-served landmark events such as meiotic recombination, formation of the synaptonemal complex, sister chromatid cohesion, spermiogenesis during postmeiotic stages, and checkpoints and factors required for the meiotic cell cycle .

精子发生中的重要基因与一系列精子发生过程中阶段性的细胞事件密切相关,例如减数分裂重组、联会丝复合物的形成、姊妹染色体的结合、减数分裂后精子的变态以及减数分裂周期中的关键点和必需因子等。

Methods GAMP was activated with cyanogen bromide and combined with 1, 6-adipic acid dihydrazide. The prepared polysaccharide derivative GAMP-ADH was coupled to recombinant protein Pep10 with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride.

GAMP经溴化氰活化后,共价接合己二酰肼手臂,在碳二亚胺催化下与重组蛋白Pep10偶联,制备结合物GAMP-ADH-Pep10。

Phage display of recombinant human lymphotoxin mutation librariesand receptor affinity screeningContraction of conformable large molecule mutation library is pivotal for molecular evolution study in vitro. In order to extend the range of the screening region of amino acid in LT gene, several receptor binding sites of LT were mutated randomly by overlap PCR amplification with the random nucleotide primers, the R46, S106, L130 combined site-directed random mutation library and R46~A52, S106~F110, R46~A52 + S106~F110 region random mutation libraries of rhLT were constructed successfully.

二。重组人淋巴毒素噬菌体库的构建及受体亲和筛选构建适合的大容量生物大分子变异体文库是进行体外分子进化研究的关键,为了扩大LT基因序列中被筛选的氨基酸范围,我们采用含随机核苷酸序列的引物,通过Overlap PCR的方法对LT的受体结合区域进行多点随机突变,分别构建了rhLT R46+S106+L130三点随机突变组合文库和R46~A52、S106~F110、R46~A52+S106~F110区域随机突变体库。

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