宓怀风
1)分子免疫学:细胞核和内质网体中亲免蛋白的研究。2)用分子生物学与高分子化学相结合的方法研究与生物大分子相关的高分子材料:克隆蛋白质作为模板用以吸附相应天然蛋白质的分子印迹聚合物的研究
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- 姓名:宓怀风
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学术头衔:
博士生导师
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学科领域:
高分子化学
- 研究兴趣:1)分子免疫学:细胞核和内质网体中亲免蛋白的研究。2)用分子生物学与高分子化学相结合的方法研究与生物大分子相关的高分子材料:克隆蛋白质作为模板用以吸附相应天然蛋白质的分子印迹聚合物的研究
1968年南开大学化学系化学专业本科毕业;78年考入南开大学元素有机研究所杨石先,陈茹玉,李正名研究生指导小组研究生;1979年去西德留学,82年6月通过博士论文答辩,获自然科学博士(Doctor rer. nat.)学位。82.10-87.10南开大学化学系任教,负责建立生物有机化学教研室并任该室负责人之一,86年被聘为副教授。1987年获西德洪堡基金会的研究基金(Alexander von Humboldt-Foundation)在海德堡大学分子遗传研究所客籍研究员。1989-1999先后在德国海德堡德国国家癌症研究中心(DKFZ)、海德堡大学医学系人类基因研究所和德国弗赖堡大学医学系生物化学和分子生物学研究所研究员,课题组组长。1999年3月南开大学高分子研究所教授、博导,2000年9月到2003年12月任吸附分离功能高分子材料国家重点实验室主任。在Molecular Cellular Biology、 Journal of Biological Chemistry、Journal of Molecular Biology、Chemical Communication、FEBS Letters、European Journal of Biochemistry、Macromolecules Human Reproduction、New Journal of Chemistry、Analytica Chemica Acta 和中国科学等国内外一流学术刊物上发表论文五十多篇,编著《生物化学》参考教材一本(南开大学出版社1990年出版)。目前作为项目负责人承担两项国家自然科学基金面上项目:‘白介素-4启动子中衰减子结合蛋白的研究’和‘用克隆蛋白质作为模板用于吸附相应天然蛋白质的分子印迹聚合物的研究’。从事的重点研究方向:1)分子免疫学:细胞核和内质网体中亲免蛋白的研究。2)用分子生物学与高分子化学相结合的方法研究与生物大分子相关的高分子材料:克隆蛋白质作为模板用以吸附相应天然蛋白质的分子印迹聚合物的研究。
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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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【期刊论文】Use of duplex probes simulating TaqMan to detect hepatitis B virus
宓怀风, De-Ming Kong, a, Yan-Ping Huang, Hui Li, Han-Xi Shen*a and Huai-Feng Mi ab
New J. Chem., 2003, 27, 721-726,-0001,():
-1年11月30日
A novel method for duplex probes is designed to simulate the TaqMan probe during polymerase chain reaction(PCR). In this method, two partly complementary single-labelled oligonucleotide probes labelled with afluorophore or a quencher, respectively, are used. At lower temperature the two probes can bind to each otherand form a mismatched duplex, in which the fluorophore and quencher are in close proximity and the sameenergy transfer mechanism as in molecular beacons may occur between them; thus, a quenching efficiencybetter than conventional TaqMan probes is acquired. In the anneal-extend step of PCR, one single-labelledprobe hybridises to the predetermined target and is cleaved by Taq DNA polymerase. Increased fluorescentsignal can be observed at lower temperature. The fluorescent data analysis demonstrated that a significantlyhigher level of fluorescent signal and hence higher sensitivity of detection is obtainable using our duplex probesin place of conventional TaqMan probes. Combined with real-time PCR instruments, the assay can be used toquantify the input target molecules and the dynamic linear range is of at least six orders of magnitude.
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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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【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】A modified molecular beacon combining the properties of TaqMan probe
宓怀风, De-Ming Kong, a, Long Gu, Han-Xi Shen*a and Huai-Feng Mi ab
CHEM. COMMUN., 2002, 854-855,-0001,():
-1年11月30日
A modified molecular beacon that possesses a stem-hairpinstructure as seen in conventional molecular beacons and canbe cleaved during PCR is designed, and it can specificallyrecognize the presence of the target and was obviously moresensitive than conventional molecular beacons.
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【期刊论文】Simulation of TaqMan by two single-labelled probes
宓怀风, De-Ming Kong, a, Long Gu, Han-Xi Shen*a and Huai-Feng Mi ab
CHEM. COMMUN., 2002, 2584-2585,-0001,():
-1年11月30日
A novel method for duplex probes is designed to simulate theTaqMan probe during polymerase chain reaction (PCR); two single-labelled probes are used for this method, whichrelies on the 5'-exonuclease activity of nucleic acid polymerase.
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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】PCR hot-start using duplex primers
宓怀风, Deming Kong, , *, Hanxi Shen, Yanping Huang & HuaifengMi
Biotechnology Letters 26: 277-280, 2004.,-0001,():
-1年11月30日
A new technique of PCR hot-start using duplex primers has been developed which can decrease the undesirableproducts arising throughout PCR amplification thereby giving better results than a manual hot-start method.
duplex primers,, hot-start,, PCR
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【期刊论文】Duplex probes: a new approach for the detection ofspecific nucleic acids in homogenous assays
宓怀风, De-Ming Kong a, Yan-Ping Huang a, Xiao-Bin Zhang a, Wei-Hong Yang c, Han-Xi Shen a, *, Huai-Feng Mi a, b
Analytica Chimica Acta 491 (2003) 135-143,-0001,():
-1年11月30日
A novel method for the detection of specific nucleic acids in homogenous solution was developed. The method is based onthe use of duplex probes in which fluorescent donor and quencher labeled on either oligonucleotide are held in close proximity, so that fluorescence is quenched. Amplification of the target sequence results in the cleavage of the probe and the resultingfluorescence can be detected. The fluorescent data analysis demonstrated that the duplex probes can specifically recognize thepresence of target, and a significantly higher lever of relative fluorescent signal than TaqMan probes is obtainable. Combinedwith real-time PCR instruments, the assay can be used to quantify the input target molecules. As few as five copies of initialtarget molecules can be detected, and a large dynamic linear ranger (five orders of magnitude) is obtained.
Duplex probe, TaqMan, Detection of specific nucleic acids
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