Publications by authors named "Jun-Chuan Lei"

Aim: Analyze the effect of Tetracycline-controlled system on Hepatitis B virus core promoter (Cp) activity and tissue specificity.

Methods: The 7 Teto sequence was amplified from plasmid pTL-8 using polymerase chain reaction (PCR) and cloned into T vector pMD19-T Simple. After sequencing, the 7 teto sequence was subcloned into upstream of Cp in pGL3-Basic/Cp.

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Objective: To explore the effect of DNA/MVA combined immunization in enhancing antibody response to MSP1.

Methods: DNA vaccine and recombined MVA were constructed based on synthesized MSP1 gene (3D7). BALB/c mice were primed with DNA solely or together with GM-CSF expressing plasmid and then boosted with rMVA/ 190.

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Objective: To explore the effect of cytokine encoding plasmids on DNA immunization in mice.

Methods: Prototype DNA vaccine VR1020/E which contain Plasmodium falciparum apical membrane antigen 1 (AMA1) ectodomain was constructed, and eukaryotic expression vectors pcDNA3/GM-CSF, pcDNA3.1(-)/IL-4, pIL-12 and pGM-CSF/pTPA-E were also built.

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Objective: To construct tetracycline operator (TetO) modified glycophorin binding protein 130 gene (GBP130) promoter of Plasmodium falciparum and investigate the position effect of insertion of TetO on the promoter activity.

Methods: Cloning of 7-copy of TetO (7cot) sequence into 4 points relative to transcriptional initiation site of GBP130 promoter in pGBPCATdelta2 plasmid (2 upstream and 2 downstream), respectively, produced 4 derivative plasmids, pG/7T (-5), pG/7T (-2), pG/7T (+2) and pG/7T (+5). After transient transfection, the expression level of reporter gene CAT in both pGBPCATdelta2 and its derivative plasmids was detected and analysed by CAT ELISA.

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Aim: To construct regulable DNA vaccine against Plasmodium falciparum by using tetracycline(Tet) regulable system.

Methods: Eukaryotic expression vectors pTL-8/apical membrane antigen 1 (AMA-1) (tTA) and pTL-8/AMA-1(rtTA) gene which express trans-activator (tTA) or reverse trans-activator(rtTA), respectively, and AMA-1 gene of Plasmodium falciparum were constructed. BALB/c mice were immunized with these plasmids and doxycycline (dox) was administered to regulate the expression of AMA-1.

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Objective: To determine the role of putative apical membrane antigen (AMA)1 domains in inducing protective immunity and to provide basis for selection of vaccine applicable segments.

Methods: Encoding gene segments of AMA1 were amplified and cloned into pET prokaryotic expression vectors. Recombinant proteins were expressed and purified.

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