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METHODS: Subtractive cDNA DNA library of PDLF was constructed with a recently developed gene cloning technique that is based on PCR and subtractive hybridization and then screened by restriction analysis and reverse southern dolt blot.

采用基于PCR和消减杂交的基因克隆技术构建体外培养的人PDLF与GF差异表达基因的扣除文库,采用酶切和反向杂交的方法筛选目的克隆,并进行测序和计算机分析。

On the basis of the previous work, we make further study on the mechanism of the denucleation process, especially on the role of different cytoskeletal elements in the denucleation process. To clone and sequence a gene related to terminal differentiation of erythroblast, and construction of two cDNA libraries of erythroblast differentiation. The results of our study are as follows

本文在我室前期工作的基础上,对红系细胞终末分化中的排核机理作了进一步的探讨,克隆、测序并表达了一个与红系细胞终末分化相关的基因,另外,还成功地构建了小鼠终末分化和差减cDNA两个红系分化cDNA文库,为本室以后的基因克隆工作打下了基础。

A 609 bp DNA sequence of DnaJ-like gene was successfully cloned from halophyte, Salicornia Bigelovii Torn, during early stage of salt stress based on suppression subtractive hybridization and comfirmed by BLAST x/n analysis.

从盐生植物海蓬子的抑制消减杂交文库中克隆了DnaJ-Like基因的长609bp的DNA序列,编码1个202aa的开放阅读框,Northern Bloting显示该基因在检测子(200 mmol/L NaCl)中的表达显著增强。

Targe genes can be selected from cDNA library, and used to analyze the expression of the genes.

从cDAN文库中可以筛选到目的基因,并直接用于该目的基因的表达。

Then Dot blot assay was used to select for and amplify differentially expressed cDNAs. Total 22 differential by displayed fragments were cloned and sequenced.

在SSH-NSC富集差异基因的基础上,我们采用了斑点杂交的方法对2个文库进行了差异基因的进一步筛选,共获得22个差异片段,克隆并测序。

In order to clone AVR-Pia, the flanking sequence of MS220K4 was used as a probe to screening the TAC library and 17 positive clones were obtained. Further restriction enzyme mapping and Southern blot analysis on 10 of the 17 positives clones narrowed the avirulence gene to a region of 10 kb.

为克隆AVR-Pia基因,以MS220K4的侧端序列为探针筛选稻瘟菌基因组TAC文库,得到17个阳性克隆,通过对其中10个进行的限制性内切酶分析并结合Southern杂交确定了目的基因所在区域。

The mouse embryonic stem cells cDNA library was screened by yeast two-hybrid assay,some candidate molecules were get,including DDX39 DEAD (Asp-Glu-Ala-Asp box polypeptide 39, BAT1A(HLA-B-associated transcript 1A), NAC1 ( nucleus accumbens-1 ), BicD2 (bicaudal D homolog 2)、MLL3 (myeloid/lymphoid or mixed-lineage leukemia 3). NAC1 and MLL3 were associated with regulation of gene transcription. DDX39 andBAT1A were associated with pre-mRNA processing and mRNA export from nucleus to cytoplasm. BicD2 was associated with the regulation of Golgi body.

通过利用酵母双杂交方法,以小鼠全长Plk1作为诱饵蛋白筛选小鼠胚胎干细胞cDNA文库中与其相互作用的蛋白,获得了DDX39DEAD(Asp-Glu-Ala-Aspbox polypeptide 39、BAT1A(HLA-B-associated transcript 1A)、NAC1(nucleus accumbens-1)、BicD2(bicaudal D homolog 2)、MLL3(myeloid/lymphoid or mixed-lineage leukemia 3)等几个候选蛋白,其中NAC1、MLL3与基因的转录调控有关,DDX39、BAT1A与mRNA前体的剪切、加工及mRNA从细胞核转运到细胞质紧密相关,而BicD2与高尔基体的调控紧密相关,提示我们Plk1与基因转录、mRNA前体的剪切、加工、mRNA的运输及胞质分裂中高尔基体的调控有关。

The study was conducted to construct the cDNA library of Chimonanthus praecox flower and analyze the expression of LTP cDNA.

构建蜡梅花cDNA文库,分析蜡梅脂转移蛋白基因的表达,为探索蜡梅冬季开花分子机理,分离克隆特色基因奠定基础。

To elucidate whether Era is related to the death of bacterial cells expressed YggG294, A double promoter expression vector that can express YggG294 and Era proteins controllably in cells was constructed.

yggG是从大肠杆菌全基因组文库中钓取并克隆的Era结合蛋白基因,研究表明该基因可能表达产物YggG294蛋白对宿主菌的生长具有强烈的抑制作用。

Full-length cDNA of Gibel carp pou2 gene was cloned from gastrula SMART cDNA library through RACE-PCR. It consists of 2421 bp, with an ORF of 1416 hp encoding a protein of 471 aa. The complete amino acid sequence shares 91.0% identity with zebrafish pou2 gene product.

用RACE-PCR方法从原肠期SMART文库中扩增到银鲫pou2基因的全长cDNA,其全长为2421bp,开放阅读框为1416bp,编码471个氨基酸,与斑马鱼pou2基因的氨基酸序列一致性高达91.0%。

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