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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。

And obtained recombinant plasmid PGEM-VP3 by linking prokaryotic expressionby vecto r PGEM-4T-1,and identified by PCR、double restriction enzyme analysis and sequence analysis,the result showed that it's true.

用EcoR I和XhoⅠ双酶切该片段并回收该片段,将此基因片段克隆至相同酶切回收后的PGEM-4T-1原核表达载体中,获得重组质粒PGEM-VP3,经PCR鉴定,限制性内切酶双酶切分析,证实了重组质粒的正确性。

A PCR product was amplified using RT-PCR method from total RNA from the abdomens of R strain using the degerated primer. The PCR product was purificated and cloned. The plasmid is pBluescript Ⅱ KS. Recombined plasmid DNA was identified by endoenzyme. The results indicated there were different P450 genes in the PCR product.

运用这对简并引物首次从德国小蠊抗拟除虫菊酯的品系R中扩增出的特异性的DNA片段,经纯化、连接和转化,重组至pBS质粒,筛选阳性克隆,提取重组质粒的DNA,并经初步的酶切鉴定,发现存在着不同的CYP4基因。

After thermal induction, no specific recombinant protein band in SDS-PAGE was found, but G-CSF activity was detectable. Therefore, a new recombinant plasmid pBVHG2 expressing hG-CSF hybrid protein with additional 8 amino acids which could be cut off specifically with the help of mucosal enterokinase at the N terminus of hG-CSF was constructed.

含此质粒菌株虽然表达菌体裂解后可测得明显的生物学活性,但SDS-PAGE仍未见特异表达产物带;因此,再应用相同方法,在hG-CSF cDNA突变体5′端增加24核苷酸对的FLAG肽编码序列,构建了hG-CSF杂交蛋白(hG-CSF天然蛋白N末端增加8aa的FLAG肽,后者可由肠激肽酶切除)的表达质粒pBVHG2。

Methods The BHD vector and the small interfer RNA targeting BHD was constructed and transfected into the cell line A549. The expression of BHD was examined by immunofluorescence. Transwell test was used to detect the invasion ability of A549 cells. Results After transfected BHD vector into A549 cells, immunofluorescence showed the expression of Folliculin located in cellular plasma.

制备BHD真核表达质粒,转染肺腺癌细胞,用免疫荧光法检测Folliculin蛋白在肺癌细胞中的表达位置;筛选sRNAi片段,选取有效抑制片段转入肺癌细胞,与转入BHD质粒的肺癌细胞对比,用Transwell实验了解BHD基因对肺癌细胞运动方面的影响。

Lividans. Using pIJ903 bearing tsr gene as a vector, the two furthermost fragments of the cluster, 4. 0kb and 1. 5kb in size respectively, were inserted into it. To facilitate the detection of gene replacement, apr gene was placed in the middle of the two inserts. OriT from E coli plasmid RK2 was also incoporated into the vector, therefore, pIJ903 derivatives can be mobilized from E.

在具有硫链丝菌素抗性基因的pIJ903的单—BamHI位点,同向插入了FR-008生物合成基因簇两个最远端的4.0kb和1.5kb片段,并在二者之间插入了可在链霉菌FR-008和变铅青链霉菌中表达的阿泊拉霉素抗性基因,同时也插入了有助于将质粒引入链霉菌的oriT片段,从而构建出了置换整个基因簇的基因置换质粒pHZ691。

Result: 1.(1) Three strips come to appear in the position of 6.0kb, 5.4kb and 600bp after gelose electrophoresis and their size accord with recombined plasmid DNA, pcDNA3 plasmid and VEGF165 gene accordingly.(2) Pure degree of the pcDNA3-VEGF165 recombined plasmid is: OD260/OD280=1.87. Its

在进行了重组质粒的扩增、提取、纯化、鉴定、兔MSCs的原代及传代培养以及RT-PCR、Western Blot 转染检测后,实验取得了令人满意的结果,证实了MSCs 能在体外有效转录外源VEGF 基因并分泌出VEGF 蛋白,同时也说明pcDNA3-VEGF165重组真核表达质粒是有效的。

Using modern biology research technique and means,plasmid extract and agarose gel electrophoresis of molecular biology level explore test were carried out for coal with not coal department the hereditary property of Thiobacillus ferrooxidans.

利用现代生物学研究方法和手段,对煤系与非煤系氧化亚铁硫杆菌的遗传特性进行了质粒抽提和琼脂糖凝胶电泳等分子生物学水平的基础性探索试验,对于质粒在硫杆菌中普遍存在的观点提出了质疑。

The isoform hMAM cDNA consisting of 270 bp, was different from the wild type one of 279 bp due to nine continuous base pair missing. Therefore the translational product of the isoform was seen to be lack of three continuous amino acid (-79~-81, VFM) compared with that of the wild type. The homology of the wild type and the isoform are testified to be 96% in both DNA and amino acid sequences.

乳腺癌组织和乳腺癌细胞株中发现两种hMAM cDNA亚型:hMAM和hMAM,长度分别为279和270bp,二者相差9个连续碱基,其翻译产物相差3个连续氨基酸残基(-79~-81,VFM),二者基因和氨基酸序列匹配度均为96%。cDNA片段分别插入原核表达质粒pGEX-KG,构建的质粒命名为pGEX-KG/hMAM和pGEX-KG/hMAM。

A pair of primers that contained flanking hydropathic amino acid codons were synthesized, to amply the sequence coding for 10~34 aa in the precursor sequence. The amplified fragment was inserted into pRSET-A to generate the first double repeat of the precursor gene. By utilizing a pair of isocaudamer BamH I and Bgl II sites, and another downstream Hind III site of plasmid pRSET-A, following a series of simple double digestions and ligation of the resulted products, a series of repeat (3, 4 and 6) precursor peptide fragment genes were derived.

本实验设计一对两侧含编码疏水性氨基酸密码子的引物,经过扩增前导序列10~34aa基因序列,并重新克隆入质粒pRSET-A构建串联二聚体后,再利用质粒pRSET-A的BamH I / Bgl II同尾酶克隆位点,经一系列简单的酶切和连接,快速构建这一前导肽中不含组氨酸标签序列的串联多聚体基因,并成功表达其六聚体重组蛋白。

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