Biomedical subjects

Yue-Ting Gong

Publications and source records attributed to Yue-Ting Gong.

4 recordsLinked to original sources

PubMed1998

Nuclear Factors Enhance the Binding of Estrogen Receptor to Estrogen Response Element.

We obtained the full length of human estrogen receptor (hER) through the in vitro translation. It was shown that the translated product could bind to the estrogen response element (ERE). The nuclear extract prepared from the rat uterus after ovariectomy could enhance the binding of hER-ERE in an estrogen dependent manner. However, the enhancing effect was sharply decreased when the nuclear extract was pre-incubated at 50 degrees for 15 minutes before being used for the binding reaction. These results indicated the presence in the rat uterus extracts after ovariectomy of a heat labile factor that can enhance the binding of hER-ERE in an estrogen-dependent manner. The DNA binding domain of estrogen receptor (ER DBD) fused to the Scistosoma japonicam glutathione S-transferase (GST) was expressed in the E. coli. The expression product also could bind the ERE. However, the binding was not affected by the uterine extract, indicating that the heat-sensitive nuclear factors may interact with hER outside the DBD to enhance the binding of ER to ERE.

Journal Article↗

PubMed1998

Fluorescence-based Quantitative Analysis for mRNA in RT-PCR Process.

A DNA fragment that forms the template of a specific mRNA can be synthesized by reverse transcription PCR. In due number of cycle, the concentration of PCR product increases proportionally to that of the template cDNA, i.e., the corresponding mRNA in RT. Since the fluorescence quantum yield of ethidium bromide greatly increased when it was inserted into the DNA double helix, the extend of expression of mRNA can be calculated by determining the fluorescence intensity under selected excitation and emission wavelengths of PCR cDNA after a proper number of PCR cycle.

Journal Article↗

PubMed1998

Participation of Water in the Binding of Estrogen Receptor with Estrogen Responsive Element in vitro.

Many reports have showed that bound water was involved in the interaction between/among the macromolecules. However, it has not been reported whether bound water is also involved in the binding of trans-factors and cis-elements in the regulation of the eukaryotic gene trans-cription or not. Preliminary studies have been made on the effect of bound water on the binding of estrogen receptor with estrogen responsive element in vitro. In the gel retardation assay using the cytosol extract of rat uterus as the supplier of estrogen receptor and 32 bp oligonucleotide containing a concensus vitellogenin A(2) ERE as the probe, various cosolvents, such as glycerol, sucrose, N-dimethylformamide and dimethylsulfoxide, were added respectively to the reaction mixture in varying concentrations to regulate the osmotic pressure. The results indicated that the binding of ER-ERE was enhanced with the increase in the final concentration of these individual cosolvents. On the other hand, when the reaction was carried out under an increasing hydrostatic pressure, the ER-ERE binding was decreased sharply. After decompression the binding of ER-ERE was gradually restored to the normal level with the lapse of time. These results suggested that bound water was directly involved in the binding of ER-ERE and may play an important role in the regulation of the eukaryotic gene transcription.

Journal Article↗

PubMed1996

The Expression of HCG Epitope Fused to Hepatits B Virus Core Antigen.

The DNA fragments encoding HGC-beta-37-CTP was amplified by PCR and fused to the core gene of HBV at the position of amino Acid 1(N-terminal fusion, pCn-HCG), 154(C-terminal fusion, pCc-HCG), 75-83(internal fusion,pCm-HCG), and both 75-83 and 154 (pC-HCG2) respectively. The fused genes were expressed in E. coli, and the antigenicity of both HBcAg and HCG as well as the expression level were analyzed. In addition, the chimeric particular characteristics of the proteins and their immunogenicity were identified. It revealed that the fusion proteins pCm-HCG and pCc-HCG were able to form particles, and that the fusion protein pCm-HCG could induce antibody of anti-HCG of high titers in mice, suggesting that the position at 75-83 amino acid residue should be a relatively promising fusion site for HCG.

Journal Article↗