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Biomedical subjects

Yuan Guan

Publications and source records attributed to Yuan Guan.

3 recordsLinked to original sources

3D-ordered macroporous materials comprising DNA.

Macroporous materials comprising DNA were fabricated with the colloidal crystal template. First, DNA and diazoresin (DR) molecules are fully filled into the voids of a colloidal crystal template. After thermal treatment and removal of the colloids, DNA porous materials with highly ordered structure were obtained. In the process of thermal treatment the cross-linking reaction takes place between DR and DNA, which plays an important role for sustaining the porous framework. The DNA porous materials will turn into a fluorescent DNA/dye composite after staining with Hoechst 33258 (Hoe), a characteristic fluorescent dye for DNA. This kind of composite DNA porous material may have potential applications in optical devices.

Animals↗

Imidazole as a catalyst for in vitro refolding of enhanced green fluorescent protein.

Imidazole is a reagent widely used in protein purifying processes. Here, we reveal a novel chaperone-like activity for imidazole using enhanced green fluorescent protein (EGFP) as a model protein. Experimental results showed that imidazole acted as an effective catalyst for refolding of the chemically denatured EGFP and suppressor for the heat-induced aggregation of EGFP. The refolding kinetics was determined in real time. Both the recovering yield and refolding rate of denatured EGFP in the presence of imidazole were increased. The studies on elucidating the mechanism show that imidazole may catalyze the prolyl cis/trans isomerization and the possible mechanism was discussed. To our knowledge, there are no data on the effect of imidazole on protein folding. Considering the prolyl isomerization is the rate-limited step for refolding of most proteins and aggregation is a universal serious problem for biotechnology, imidazole thus represents a previous unknown type of protein-folding catalyst.

Catalysis↗

Universal molecular beacon-based tracer system for real-time polymerase chain reaction.

DNA diagnostic has been moving from expensive, low-throughput, multistep methods to inexpensive, higher throughput, closed-tube, and automated methods. Fluorescence is the favored signaling technology for such assays. In this method, we describe a universal molecular beacon (U-MB) as the fluorescent tracer in the real-time PCR technique. A 5'-universal template primer (5'-UT primer) has been designed with a tail in complementary to the loop and 5'-side arm sequence of U-MB at the 5'-end of forward target specific primer. As PCR cycles increase, a new DNA fragment with a 5'-UT primer tail is synthesized, which is used as the template for next PCR cycle. As the reverse primer extends to the 5'-UT primer tail, the U-MB hybridized is displaced and the fluorescence from the fluorophore of the U-MB is quenched, indicating that the allele-specific PCR is in progress. This tracing system combined with an allele-specific reverse primer and vent (exo-) DNA polymerase, a polymerase that lacks 3'- to 5'-exonuclease activity, was used for the detection of point mutations of base G in codon 259 (AGA) of exon 7 of p53 gene on a panel of breast cancer individuals.

Breast Neoplasms↗