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【期刊论文】Real-time quantitative assay of telomerase activity using the duplex scorpion primer
宓怀风, Yanping Huang, , Deming Kong, Yi Yang, Ruifang Niu, Hanxi Shen, * & HuaifengMi
Biotechnology Letters 26: 891-895, 2004.,-0001,():
-1年11月30日
A novel real-time quantitative method for measuring telomerase activity is described in which the duplex scorpionprimer is used to provide an intramolecular probing mechanism for specific detection of telomerase activity. Usingthis method, linearity from 10 to 104 cells expressing telomerase activity could be obtained (R2=0.994). Therequirement of post-PCR steps is thus obviated.
duplex scorpion primer,, real-time quantitative PCR,, telomerase activity,, TRAP assay
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【期刊论文】Detection of Hepatitis B Virus DNA by Duplex ScorpionPrimer-based PCR Assay†
宓怀风, KONG, De-Ming*, a, b, SHEN, Han-Xi b, MI, Huai-Feng a
Chinese Journal of Chemistry, 2004, 22, 903~907,-0001,():
-1年11月30日
The application of a new fluorogenic probe-based PCR assay (PCR duplex scorpion primer assay) to the detectionof Hepatitis B virus (HBV) DNA in human sera was described. Duplex scorpion primer is a modified variant ofduplex Amplifluor, and the incorporation of a PCR stopper between probe and primer sequences improve the detectionspecificity and sensitivity. Combined with PCR amplification, this probe can give unambiguous positive resultsfor the reactions initiated with more than 20 HBV molecules. In addition, the particular unimolecular probingmechanism of this probe makes the use of short target-specific probe sequence possible, which will render thisprobe applicable in some specific systems.
duplex scorpion primer,, hepatitis B virus (, HBV), DNA,, fluorogenic probe
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【期刊论文】Protein Interactions of the MLL PHD Fingers Modulate MLL Target Gene Regulation in Human Cells
宓怀风, KERI FAIR, , MELANIE ANDERSON, ELENA BULANOVA, HUAIFENG MI, MAXIMILIAN TROPSCHUG, AND MANUEL O. DIAZ*
MOLECULAR AND CELLULAR BIOLOGY, May 2001, p. 3589-3597,-0001,():
-1年11月30日
The PHD fingers of the human MLL and Drosophila trx proteins have strong amino acid sequence conservationbut their function is unknown. We have determined that these fingers mediate homodimerization andbinding of MLL to Cyp33, a nuclear cyclophilin. These two proteins interact in vitro and in vivo in mammaliancells and colocalize at specific nuclear subdomains. Overexpression of the Cyp33 protein in leukemia cellsresults in altered expression of HOX genes that are targets for regulation by MLL. These alterations aresuppressed by cyclosporine and are not observed in cell lines that express a mutant MLL protein without PHDfingers. These results suggest that binding of Cyp33 to MLL modulates its effects on the expression of target genes.
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宓怀风, Xuejun Sun, Suxia Chen, Shunzi Li, Husheng Yan*, Yunge Fan, Huaifeng Mi
Peptides xxx (2004) xxx-xxx,-0001,():
-1年11月30日
In our previous paper it was shown that the two C-terminal Gln residues of a C-terminal 15-residue fragment, Mel(12-26)(GLPALISWIKRKRQQ-NH2), of melittin and a series of individual substituted analogues might not involved in the interaction withbacterial membranes. In this paper, peptides with one and two Gln residues deletion, respectively, Mel(12-25) and Mel(12-24), weresynthesized and characterized. Both of the deletion peptides showed higher antimicrobial activities than the parent peptide, Mel(12-26). If both of the Gln residues of Mel(12-26) were respectively replaced by a hydrophilic amino acid Gly, the antimicrobial activity increasedslightly. If the Gln residue of Mel(12-25) was replaced by a hydrophobic amino acid Leu, the antimicrobial activity changed little, althoughthe substituted peptide possessed much higher hydrophobicity and higher α-helical conformation percentage in 1,1,1,3,3,3-hexafluoro-2-propanol/water determined by circular dichroism spectroscopy (CD) than the parent peptide. These results indicated that the two C-terminalresidues might be indeed not involved in the binding to bacterial membranes. The antimicrobial activity increasing with the residue deletionmay be caused by the decrease of the translational and rotational entropic cost of the binding of the peptides to bacterial membranes becauseof the lower molecular weights of the deletion peptides.
Melittin, Antimicrobial peptide, Antibiotic, Hemolysis
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宓怀风, Yan-Ping Huang, ab, De-Ming Kong, a, Qi-Meng Chen, b, Han-Xi Shen*a and Huai-Feng Mi ac
New J.Chem., 2004, 28, 1488-1493,-0001,():
-1年11月30日
Telomerase, a ribonucleoprotein enzyme, is expected to be a new marker for cancer diagnosis. Thedevelopment of the telomeric repeat amplification protocol (TRAP) and its modified versions have facilitatedthe detection of telomerase activity in small tissue and tumour samples. But most of these techniques requirecomplex post-PCR procedures. As for the two real-time quantitative methods (SYBR Green and Amplifluormethods) reported so far, both use fluorogenic probes without specificity. To overcome these problems wedeveloped a new real-time method for the detection of telomerase activity. In this method a duplex scorpionprimer and reverse primers with hairpin-like structures were used. The use of duplex scorpion primers showsa series of advantages: the target-specific probe sequence provides higher specificity than the SYBR Greenand Amplifluor methods; the unimolecular probing mechanism allow the assay to be conducted under fastcycling conditions and a single operation can be completed in 1.5h; the closed-tube system reduces the riskof carryover contamination and supports high throughput. This method may be a useful tool to rapidly,specifically and precisely quantify telomerase activity.
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