1.Construction of HPV18E7 recombinant plasmid and exploration of its optimization expression condition in Escherichia coli
Renjian HU ; Jiali CAI ; Li LIU ; Manyu TU ; Tao XU ; Cuirong DU ; Jia LUO ; Sen DING
Chongqing Medicine 2013;(30):3647-3649
Objective To construct recombinant plasmids containing HPV18E7 gene ,and explore the optimization condition of its expression in Escherichia coli .Methods The genomic DNA extracted from HeLa cell line which served as a template to the HPV18 E7 gene was amplified using PCR method ;and the amplified product of HPV18E7 gene was connected to the pET-32a(+ ) vector ,which composed the pET-32a(+ )-HPV18E7 recombinant plasmid ;the positive recombinant plasmids were transformed into BL21-DE3-pLysS competent cells and the optimized expression condition was explored in order to obtain a large amount of HPV18E7 oncogenic protein .Results The fragment length of PCR products of HeLa cell genomic DNA was consistent with that of HPV18 E7 gene .In LB medium ,the expression level of the target protein was not high under such conditions as different concentra-tion of IPTG and lactose ,different temperatures and different induction starting amount .Therefore the ZYM-5052 auto-induction medium was tried in this experiment ,and the expression amount of the fusion protein was much higher than that induced with IPTG and lactose .Conclusion The amount of HPV18E7 fusion protein in ZYM-5052 automatic induction medium is much higher than that induced with IPTG and lactose .
2.Synthesis, characterization and blood compatibility studies of biomedical aliphatic polyurethanes.
Minhui DU ; Jianshu LI ; Yang WEI ; Xingyi XIE ; Chengsheng HE ; Cuirong FAN ; Yinping ZHONG
Journal of Biomedical Engineering 2003;20(2):273-276
The one-step method was adopted in this study to synthesize aliphatic polyurethane with 4,4-methylene dicyclohexyl diisocyanate(HMDI), 1,4-butanediol (BDO) and poly (tetrahydrofuran) (PTMG). The tests conducted on this material were: FIR spectrum, mechanical properties test, water contact angles test, hemolysis test and platelet adhesion test. Results showed that this material has a good tensile strength up to 30 Mpa, similar to aromaphatic polyurethane. But its tensile elongation, tensile permanent change, hydrophility are better than those of aromaphatic polyurethane. The hemolysis test and platelet adhesion test showed that it has good blood compatibility.
Animals
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Biocompatible Materials
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chemical synthesis
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chemistry
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toxicity
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In Vitro Techniques
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Materials Testing
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Platelet Adhesiveness
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drug effects
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Polyurethanes
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chemical synthesis
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chemistry
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toxicity
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Rabbits
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Spectroscopy, Fourier Transform Infrared
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Surface Properties
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Tensile Strength
3.Synthesis, characterization and blood compatibility studies of waterproof breathable polyurethanes.
Peng WANG ; Jianbin LUO ; Minhui DU ; Chengsheng HE ; Cuirong FAN ; Yinping ZHONG
Journal of Biomedical Engineering 2005;22(4):734-738
Adopting the two-step method and changing the proportion between PEG (Polyethylene glycol) and PTMG (poly (tetrahydrofuran), we used the MDI (4,4'-diphenylmethane diisocyanate) and short chain extender BDO (1,4-butanediol) as hard segment, the PTMG and PEG as soft segment, and hence prepared a series of polyether-based thermoplastic polyurethanes. FTIR showed the structure character of these polyurethanes. The determination of mechanics property and water contact angles revealed their good mechanics properties and hydrophilicity. Blood compatibility was evaluated by hemolysis test and platelet adhesion test, which revealed their good hemocompatibility. So those polyurethanes may be of wide application in the future.
Animals
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Biocompatible Materials
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chemical synthesis
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chemistry
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Butylene Glycols
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chemistry
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Isocyanates
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chemistry
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Materials Testing
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Polyethylene Glycols
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chemistry
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Polymers
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chemistry
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Polyurethanes
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chemical synthesis
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chemistry
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Rabbits