When 2 μg, 5 μg, 10 μg of pSG5UTPLHBX DNA were transfected together with reporter gene pHEC 2×CAT in the HepaG2 cells, the CAT activities were (45.5 ±25.9)%, (65.6±14.4)%, and (68.8 ±19.7)%, respectively.
the role of TRAF2 in LMP1, LMP1 (1-231) or LMP1 A187-351-mediated NF-kB activation was evaluated using TRAF2 dominant negative mutant TRAF2△6-86 with reporter gene analysis;
Objective To study the effect of inactivated Coxsackie virus B1(CVB1), which was considered to have high affinity to skeletal muscle cells, on gene transfer efficiency of reporter gene, pCDNA3LacZ, in primary muscle cells.
The 5′flanking sequence of this gene has been cloned with genomic walker kit, and its promoter activity was confirmed through luciferase report gene test.
The hTERT expression and telomerase activity were respectively tested with SEAP report gene assay and with PCR-ELISA analysis in human nasopharyngeal epithelial cells.
Resul ts: After sequence analysis and homology comparison, the candidate p r otein was identified as PRO2605. KGF-2 and PRO2605 could activate the transcrip tion of report gene.
Application of Modern Biotechnology to Environmental Microbiology: Ⅲ.Restriction Fragment Length Polymorphism Analysis?Denaturing/ Temprature Gradient Gel Electrophoresis and Reporter Genes
Aim:To investigate the expression of GFP reporter genes transfected in rat C6 glioma cells as a tag and its effect on biological characteristics of tumor cells.
In this report, the results of immunocytochemistry and Western blot analysis showed that the active form of Stat3 protein existed in COS7 cells. The endogenous Stat3 protein could also induce the expression of m67 sequence directed reporter genes.
Results The high concentrations of newly synthesized RNA were accumulated in cytoplasm when tripartite leader sequence was present in reporter RNA, despite the equal rates of transcription of the two reporter genes.
The sequencing results were aligned with the sequence (Genbank, AY306198, EF012241, L77887, M81327, M92089), indicating that the homology between these two sequences was 99%, indicated that this gene is PLF.
Next, BsNPVp35 was cloned into the MCS of pIZ/V5-His before its stop- codon was mutated and EGFP gene was cloned downstream of p35 gene, be sure that they are in the same reading frame.
Methods: Three fusion reporter plasmids named pM4CAT2、pCMVCAT、pM4CMVCAT were constructed by inserting four repeated copies of c myc specific binding squences(CACGTG) 4、CMV enhancer/promoter 、(CACGTG) 4 containing CMV enhancer/promoter into pBLCAT2 respectively.
Methods: 1.06kb fragment upstream of Nkx3.1 gene was amplified by PCR using human genomic DNA as template. Its promoter activity was determined with dual-luciferase reporter assay after it had been cloned into pGL3-basic vector and transfected into LNCaP cells.
The analysis of the GAL4 luciferase reporter gene indicated that fragments from 228 to 407 amino acids in the DUF654 domain had a strong transcription repression activity.
shown that AcNPV could be used as gene transfer vector in mammalian cells and that barnase is the suitable report gene which could be detected more easily than that commonly used.
It was concluded that the recombinant adenovirus carrying rat calcineurin A α (Ppp3ca) cDNNA as well as a report gene enhancer green fluorescent protein gene was successfully constructed in this experiment.
The HepG2 cells were irradiated by ultrasound at 1 MHz, 0.4-2.0 W/cm2 and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene.
To evaluate the optimal parameters for efficient gene transfer into human hepatoma HepG2 cells by the DNA-poly(L-Lys)Gal complexes, the bacterial reporter genes lacZ and cat were used.
In addition, GhEREB2 and GhEREB3 proteins specifically bind to a GCC-box and strongly activate the expression of HIS3 and LacZ reporter genes in yeast.
An original yeast system with two reporter genes oriented oppositely to each other was developed to study the regulatory functions of noncoding genomic sequences.
A screening of the yeast library of rat lung cDNA allowed the identification of six proteins specifically activating the reporter genes of a two-hybrid system and 21 unlikely protein partners of protein p45.
A PEA-resistant temperature sensitive mutant strain FD2003 was isolated follow-ing DES mutagenesis.Its DNA and protein synthesis,its permeability to ammo acids,bases,crystal violet and 6-APA and its sensitivity to various antibiotics and outer membrane protein-specific phages were determined.Based on these experimental results it was concluded that FD2003 carried a mutation defective in the regulation of synthesis or assembly of outer membrane proteins.By means of Plvir transduction this mutant was mapped a...
A method for amplification and purification of gene prode in Escherichia coli(E.coli)is reported in this paper.Plasmids containing gene probe were transfectedinto E.coli by treating bacteria cells with calcium chloride.The transfected bacteriawere cultured to allow plasmid amplification.Then the amplified plasmids wereextracted from E.coli by alkline lysis procedure.The results are as follows:(1)Theefficiency of transformation of plasmid is related to its concentration.(2)Aftertransformation,amplification a...
The present article reports the acute toxicity of recombmant human interferon-αB (rHuIFN-αB) in mice and its long-term toxicity in rats. The results showed that rHuIFN-αB im LD50>10 9 TU/m2 and there were no significant differences of all parameters (appearance, vitality, activity, appetite, urine, stool, body weight, temperature, blood chemistry, biochemistry and pathological findings) between the medicinal group and normal control group. So there were no toxic reactions in rats when rHuIFN-αB was given 30...
本文报道基因工程人αB干扰素(Recombinant human interferon-αB,rHuIFN-αB)对小鼠的急性毒性和夫鼠的长期毒性。rHuIFN-αB肌注小鼠急性毒性LD_(50)>10~9IU/m~2,大鼠长期毒性所有观察指标(一般行为活动、食量、粪便、体重、体温,血象、牛化指标和病榆器官等)在给药组和对照组间未见明显差别,表明大鼠肌注rHuIFN-αB相当于推荐临床剂量的30倍,连续3个月,未显示药物性毒性反应。
报道基因
reporter gene
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