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Methods HBx gene with EcoR Ⅰ and Hind Ⅲ endoenzyme sites was obtained by using PCR from plasmid pcDNA3.1-HBx.

用PCR方法从质粒pcDNA3.1-HBx中扩增含Kpn Ⅰ和Hind Ⅲ酶切位点的HBx基因序列。

In my experimentⅠcompare the purposes of different combinations of endoenzyme(MseⅠ-HindⅢ, MseⅠ-PstⅠ, MseⅠ-EcoRⅠ) and find that MseⅠ-HindⅢeasily finds thepolymorphism by primers combinations suiTab for AFLP analysis.And by means of this there aremany polymorphism strips. So the double endoenzyme of AFLP was exercisedfor the analysis of soybean cytoplasmic male sterility.In the cource of my experiment, 64 primerpairs of MseⅠand HindⅢwere exerted to analyse the twe mtDNA pools which are made ofsterilities and holdings and the polymorphisms of sterilities and holdings.

运用扩增片段长度多态性的方法,对独特的杂交大豆试材进行分析研究,通过比较MseⅠ-HindⅢ、MseⅠ-PstⅠ、MseⅠ-EcoRⅠ不同酶切组合的效果,结果表明MseⅠ-HindⅢ较容易找到多态性引物组合,而且具有较高的多态性,本试验选用MseⅠ-HindⅢ进行双酶切大豆线粒体DNA从而完成AFLP的分析工作,试验用64对MseⅠ和HindⅢ引物对两个不育系及保持系材料的线粒体DNA制成的pool进行AFLP标记分析,分析了不育系与保持系之间的多态性; 4。

Methods Genomic DNA samples of two isolated Plasmodium falciparum isolate strains prepared directly from 5 cases of cerebral malaria patients′ blood in mengla County, Yunnan Province and in Yingjiang County, Yunnan Province were used for polymerase chain reaction amplification and the two pairs of oligonucleotides for the highly conserved genes encoding FC27 merozoite surface protein 2 (MSP2) of Papua New Guinea strain of Plasmodium falciparum were used as primers. The PCR products were digested with BamH1 and Hind Ⅲ respectively, and the generated fragment MSP2 were cloned into M13mp18 and M13mp19 vectors and their DNA was analyzed as the templates for DNA sequencing by the dideoxy chain-termination method.

应用多聚酶链反应对5例中国脑型疟患者恶性疟原虫云南省勐腊县勐罕分离株和云南省盈江县农场分离株基因组裂殖子表面蛋白2(MSP2)基因进行扩增,将扩增产物分别经BamHI和Hind Ⅲ双酶切后,回收的MSP2基因分子定向克隆M13mp18和M13mp19载体,按Sanger双脱氧链终止法进行DNA序列测定,并与恶性疟原虫株FC27、K1、IC1和CAMP株进行同源性分析比较。

Methods HBV DNA was extracted from the serum of patient with hepatitis B. The X gene was amplified by PCR using the primers with EcoRI and HindⅢ digestion sites, and then cloned into pronucleus expression vector pMAL-C2X, which was detected by EcoRI and HindⅢ digestion and sequence.

从乙肝病人血清中提取HBV DNA;以带有EcoRI和HindⅢ酶切位点的引物,用PCR方法扩增X基因;而后定向克隆到原核表达载体pMAL-C2X上,经酶切鉴定和序列分析;以IPTG诱导X融合蛋白的表达。

Brahmagupta's treatise Brahma-sputa-siddhanta was translated into Arabic under the title Sind Hind.

婆罗门伽塔的论文Brahma-sputa-siddhanta被翻译成阿拉伯语,题目是Sind Hind。

Methods:The polymerasechain reactionand restriction fragment length polymorphismtechniques were used to detect both Hind III and Pvu IIpolymorphisms in 92 children with simple obesity and 80 healthycontrols.The levels of the plasma lipid,plasma lipoproteins,bodymass index,blood pressure,waistline,chest circumference,gluteal circumference,skinfolds thickness at three measuring points(biceps,subscapular and abdominal wall)were also measured.

应用聚合酶链反应和限制性内切酶片断长度多态性技术,检测了92例单纯肥胖症儿童和80例正常健康儿童的LPL-Hind III与LPL-Pvu II基因多态性,并测定血脂、BMI、血压、腰围、胸围、臀围、以及肱二头肌、肩胛下、腹壁等三个部位的皮褶厚度。

The caprine IL-2 gene and the IFN-γgene without the signal peptidesequence were subcloned into (5") Bam H I and (3") HindⅢrestriction sites of pET30aplasmid, the recombinant plasmid were transformed into E. coli BL21(DE3) and induced withIPTG, they were demonstrated by SDS-PAGE that the genes were expressed as inclusion bodies.

应用DNA重组技术,将IL-2基因和去除信号肽的IFN-γ基因分别插入到原核表达载体pET-30a的BamHⅠ和HindⅢ位点上,构建原核表达质粒pET30a-CapIL-2和pET30a-CapIFN-γ,转化大肠杆菌BL21(DE3),经IPTG诱导后进行SDS-PAGE电泳,结果表明目的基因在大肠杆菌中以包涵体形式融合表达。

Methods The target cDNA was amplified by PCR and PCR products were indigested by restriction enzyme Nco Ⅰ and Hind Ⅲ. Then the indigested products were recombined into pET-BBH and pET-BBH was expressed in E. coli BL21 (DE3) by IPTG inducing. Next, the expressed proteins were identified by SDS-PAGE, western blot and enzyme activity test. Finally, fluorescence quantitative PCR was used to test its expression quantity in different worm stages.

PCR扩增GAMMA-BBH cDNA基因,产物经NcoⅠ和HindⅢ限制性内切酶酶切后连接至原核表达载体重组为pET-BBH,在BL21(DE3)中用IPTG诱导表达,SDS-PAGE、Western blot和酶活性测定鉴定表达产物,用荧光定量PCR方法检测广州管圆线虫不同虫龄GAMMA-BBH的表达量。

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。

The Charon library DNA was then digested with restriction enzymes BamH Ⅰ and Hind Ⅲ, and DNA fragments of length of about 0.5-1. 5kb was subcloned into plasmid pUC 18 which was also digested by BamH Ⅰ and Hind Ⅲ and dephosphorylated by alkaline phosphatese before being ligated with Charon fragments.

抽提λCharon文库总DNA,用BamHⅠ和HindⅢ完全酶切电泳,用低熔点琼脂糖回收长约0.5-1.5kb的DNA片段;连接到pUC18上,pUC DNA先用BamHⅠ和HindⅢ酶切,后用碱性磷酸酯酶脱磷。

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No, this is not what we mean by framing, although the principle is the same.

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