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Although overexpression of Hspa5 does not cause detectable defects, its knockdown by injecting Hspa5 morpholino antisense oligonucleotides results in slower epiboly during gastrulation and dorsalized phenotypes at 24 hpf, including smaller and curled tail and absence of caudal ventral fin. When Hspa5 expresison in embryos is knockdown, the expression of the dorsal markers gsc and ntl is enhanced while the expression of ventral markers is reduced.

尽管在胚胎中过量表达Hspa5对胚胎发育没有明显影响,但是利用反义吗啉环寡核苷酸抑制胚胎中Hspa5的表达会导致胚胎外包变得缓慢,24hpf出现尾部变细或卷曲、缺少腹鳍等背部化的表型,同时gsc和ntl等背部标记基因的表达增强,但eve1等腹部标记基因的表达减弱。

A genechip analysis was performed using RNAs derived from embryos injected with squint mRNA, MZoep mutant embryos that are deficient in Nodal signaling, and wildtype embryos at the 30% epiboly stage Transcriptswith at least two-fold changes in expression level between wildtype and the other samples were identifyied In squint mRNA-injected embryos, 265 transcripts show anincreased expression level and 111 have a decreased expression level; in MZoep embryos, the expression of 1 495 transcripts increases while 550 transcripts express at a decreased level.

为鉴定受Nodal信号调控的基因,特别是那些转录因子基因,通过将来自squint过量表达、缺失Nodal信号的MZoep突变体或野生型30%外包期胚胎的RNA与Affymetrix斑马鱼寡核苷酸芯片杂交。

Methods The LPL cDNA was isolated from the human epiploon adipose tissue by means of RT-PCR. The LPL cDNA was ligated into the pcDNA3.1Zeo. The recombinant pcDNA3.1Zeo-LPL cDNA was identified by endonucleases, PCR and DNA sequencing. COS-1 cells were transfected with the recombinant LPL gene plasmid using Lipofectamine 20001M The LPL mass in cells and the culture medium were determined by a Markit-M LPL Kit. Spectrophotometry was used to measure the LPL activity.

采用RT-PCR法从人大网膜脂肪组织获取LPL cDNA基因,以pcDNA3.1Zeo质粒作为载体,运用基因克隆技术构建野生型人LPL cDNA重组质粒,限制性内切酶酶切、PCR以及双向测序鉴定重组质粒;运用Lipofectamine 2000将重组pcDNA3.1 Zeo-LPL cDNA质粒导入COS-1细胞,RT-PCR法检测LPL mRNA,ELISA法和比色法分别检测细胞裂解液及细胞培养基中LPL浓度及活性。

2 STR-PCR We analysed the results of STR with the silver dye in the beginning. The silver dye of STR analysis on a sporadic DMD family shows the band confirmation is subjective by eyeballing. It can not detect the small repeats and the size of polymorphic fragments. For limitation of the silver dye, we analysed the band size of STR with the SAGAGT Genotyping System. Primer are designed according to the nucleotide sequence of five STR markers (located in 5' terminus and intron 44, 45, 49, 50) of dystrophin gene. The 5' end of the forward primer are attached a M13 tailed primer which conjugated to the infrared flourescent dye.

根据dystrophin基因5个多态位点(44、45、49和50号内含子以及5'端启动子区的序列)设计引物,在这些STR的上游引物5'端加一可被红外线荧光物质IRDye700识别标记的M13fw的尾:CAC GACGTT GTA AAA CGA C 通过PCR扩增出重复序列本身,经高分辨率的变性聚丙烯酰胺凝胶电泳后,利用红外激光遗传分析系统及SAGA〓基因分型软件对凝胶上的扩增产物及内标的位置,进行扫描分析,确定产物片段的大小。

Mutation rate at exon of GH in five bovine species was very low with the percentage of 3.48%. The majority of nucleotide substitution was nonsense mutation and only one missense mutation was observed. Molecule tree based on haplotypes of bGH at exon 5 showed that differentiation was apparent relatively between Bubalus bubalis and Bos taurus, Bos indicus, Bos grunniens, Bos frontalis. There was no apparent differentiation among other four bovine species and they shared mutual ancestral sequence.

序列分析表明,5个牛种GH基因外显子5的遗传变异水平较低,平均核苷酸变异率约为3.48%,而且绝大多数位点的核苷酸替换是同义突变,仅发现1个错义突变位点;从GH基因外显子5序列单倍型构建的分子进化树来看,水牛与普通牛、瘤牛、牦牛及大额牛的分化相对比较明显,其他4个牛种之间并无明显分化,还享有共同的祖先序列。

To further understand mechanisms that control male gametogenesis and embryo development,we performed an genetic screen for mutations that caused distorted Mendelian segregation in Ac/Ds transposon system in Arabidopsis thaliana.

通过卡那霉素抗性(由突变体中标记基因编码)非孟德尔分离,从Ds插入的拟南芥突变体库中筛选到一个雄配子体发育突变体:ovp1和一个早期胚胎发生突变体eed1,我们对这两个突变体的表型和相关基因进行了研究。

Hairy roots were induced from tobacco leaves with Agrobacterium rhizogenes LBA1334 harboring agropin type PRil855 and binary vector PIG121HM carrying a kanamycin resistant gene nptll and GUS gene and the good hairy root clones were selected. The root with many root hairs and branches grew vigorously toward all the direction on MS medium without plant hormone and on kanamyain containing medium (km:30mg/L). The structure of the hairy root tip is different from that of ordinary root. Witch was not typical root cap maybe the main cause of losing the geotropism of hairy root. Plantlet regenerated from hairy root when cultured on Ms medium without plant hormone. The nicotine content of the hairy root was little higher than that in the natural roots.

本研究通过发根农杆菌工程菌LBA1334(含有野生型pRi1855质粒和pIG121HM双元载体质粒,编码NPTⅡ基因和GUS基因)转化烟草沙姆逊,获得了具有卡那霉素抗性烟草发状根,并筛选出良好的单克隆株系,该发状根具有典型的发状根特性(多根毛、多分支、在无激素培养基上快速生长),通过根尖压片分析,烟草发状根不具备典型的根冠;发状根中烟碱含量1.61%稍高于与天然栽培品种母体根的含量(1.5%左右),发状根向培养液中释放烟碱,释放量最多时占总烟碱量的81.1%;在无激素的培养基上获得了再生植株。

Different forms of a gene, which may give rise to different phenotypes, are known as alleles.

不同形式的一种基因,这可能会导致不同的表型,被称为等位基因。

Second, low glutelin content was regulated at the level of transcription, but the suppressed glutelin genes were not among those published.

采用定量RT-PCR方法对LGC-1及其野生型日本优中已知谷蛋白基因逐个进行表达分析,结果表明,GluA-1,GluA-2,GluA-3和GluB-1,GluB-2,GluB-4基因的转录水平在二个品种中无明显差异。

Gracilis were applied to investigate the effect of proplastid on adaptability, in which such structure was found to be important to the competitiveness during heterotrophic growth in the dark; 2 a new procedure for differential display technology was developed and used to study the influence of plastid on nuclear gene expression, with which it was shown that the loss of plastid could cause remarkable changes in expression of certain nuclear genes.

本文主要有两点突出之处:1)应用裸藻褪色突变株与野生型的生长竞争实验探讨了原质体对于裸藻的适应意义,发现在黑暗环境和异养生长条件下原质体对于细胞的生存竞争有重要作用;2)改进了一种差异显示技术并利用该技术来研究质体的有无对基因表达的影响,证明质体丢失可导致某些核基因表达的显著变化。

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