Level of CYP4G19 Expression Is Associated with Pyrethroid Resistance in Blattella germanica.

German cockroaches have become a large problem in the Shenzhen area because of their pesticide resistance, especially to pyrethroid. A pyrethroid called "Jia Chong Qing" to prevent pests for a long time were found to be resistant to "Jia Chong Qing" with resistance index of 3.88 measured using RT-PCR and immunohistochemistry analysis showed that both CYP4G19 mRNA and CYP4G19 protein expression levels in the wild strain were substantially higher than that of a sensitive strain.

[Prediction and modification analysis of low-affinity HLA-A*0201 restricted CTL epitopes derived from HIV-1 pol antigen].

Aim: To screen the possible HLA-A*0201 restricted low-affinity CTL epitopes derived from HIV-1 pol antigen and to predict and identify the possible change of the affinity between epitope and the HLA-A*0201 molecule when the epitope is modified.

Methods: HLA-A*0201 restricted low-affinity CTL epitopes were predicted by CTL epitope prediction software based on super motif, proteasome cleavage probability, HLA affinity and so on. The candidates were modified acid substitution and analyzed by computer.

Characterization of the genome sequence of an oncolytic Newcastle disease virus strain Italien.

We determined the complete genome sequence of strain Italien, a virulent and oncolytic strain of Newcastle disease virus (NDV) by direct nucleotide sequencing of RT-PCR products, a size of 15,186 nucleotides (nt). Comparison of six coding genes and non-coding regions of Italien with those of the other 25 sequenced strains revealed NDV Herts/33 was the most similar strain with Italien. The gene encoding the RNA dependent RNA polymerase was the most highly conserved, while the gene encoding phosphoprotein was the most highly variable.

[Cloning and expression of PbTIP analogous gene from Plasmodium berghei ANKA and preparation of its polyclonal antibody].

Objective: To clone and express a novel protein analogous to TIP (T cell immunomodulatory protein) of Plasmodium berghei ANKA and prepare its polyclonal antibody.

Methods: The PbTIP encoding nucleotide sequence was searched from the Plasmodium berghei genomic database and amplified by PCR. The gene was sub-cloned into prokaryotic expression vector pGEX4T-1 and expressed in E.

Targeted inhibition of HBV gene expression by single-chain antibody mediated small interfering RNA delivery.

Unlabelled: RNA interference is highly effective at inhibiting HBV gene expression and replication. However, before small interfering RNA (siRNA) can be used in the clinic, it is essential to develop a system to target their delivery. Antibody-mediated delivery is a novel approach for targeting siRNA to appropriate cells.

[Adenovirus-mediated delivery of targeted ribonuclease against HBV replication in vitro].

Objective: To observe the inhibitory effect of targeted ribonuclease delivered via adenovirus against HBV replication in vitro.

Methods: The shuttle plasmids pDC316 were constructed on the basis of the previous plasmids pcDNA3.1(-)/TRL, pcDNA3.

[Expression, purification and transduction of Tat-p53 fusion protein].

Aim: To express and purify Tat-p53 fusion protein and investigate its transduction efficiency.

Methods: The gene encoding wide-type p53 was isolated using RT-PCR from A549 cell line and cloned into pTAT-HA and pET32a prokaryotic expression vectors. Recombinant plasmids were transformed into E.

[DNA/MVA combined immunization: antibody response to Plasmodium falciparum merozoite surface protein 1 in mice].

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.

Effect of vector-expressed shRNAs on hTERT expression.

Aim: To study the effect of short hairpin RNAs (shRNAs) expressed from DNA vector on hTERT expression.

Methods: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expression vector pUC18U6 to form pUC18U6ht1-4, which were then introduced into HepG2 cells by using liposome-mediated transfection. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir, which expressed shRNA against green fluorescent protein (GFP), were used as controls.

Anti-HBV activity of TRL mediated by recombinant adenovirus.

Aim: To investigate the inhibitive effect of hepatitis B virus (HBV)-TRL on HBV replication.

Methods: Based on previously constructed pcDNA3.1(-)/TRL, TR, TRmut, HBV core protein (HBVc) and hEDN, interest gene sequences TRL, TR, HBVc and hEDN were inserted into adenovirus shuttle plasmid pDC316 respectively and co-transfected HEK293 cells with rescue plasmid pBHGloxdeltaE1,3Cre to acquire RAd/TRL, TR, HBVc and hEDN.

Effect of sodium citrate based anticoagulants on the growth activity of malaria parasites.

Objective: To study the effect of anticoagulants based on sodium citrate on the growth activity of malaria parasites.

Methods: The parasites were treated with 3 anticoagulants (ACD, CD and SC), respectively, and the parasitemia was determined to measure the effect of the anticoagulants on the growth of the parasites. Unsynchronized Plasmodium falciparum was treated with the anticoagulants at different concentrations for 3 h at 37 degrees C.

[Modulation of immune response to Plasmodium falciparum apical membrane antigen 1 DNA vaccine by cytokine plasmids].

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.

[Quantitation of human telomerase reverse transcriptase mRNA with real-time fluorescent RT-PCR].

Aim: To establish a real-time fluorescent RT-PCR assay to quantify human telomerase reverse transcriptase (hTERT) mRNA.

Methods: Total cellular RNA was isolated from HepG2 cells using Trizol reagent, which was then reverse-transcribed into cDNA. By using a pair of gene-specific primers and a TaqMan MGB probe, cDNA of hTERT was quantified with real-time fluorescent PCR.

Quantifying anti-HBV effect of targeted ribonuclease by real-time fluorescent PCR.

Aim: To quantify the inhibition of HBV replication by targeted ribonuclease by using real-time fluorescent PCR.

Methods: Targeted ribonuclease was introduced into 2.2.

Effects of tetracycline operator element insertion on Plasmodium falciparum glycophorin binding protein 130 gene promoter activity.

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.

Effect of vector-expressed siRNA on HBV replication in hepatoblastoma cells.

Aim: To study the effect of siRNA expressed from DNA vector on HBV replication.

Methods: Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.

[Preliminary study on regulable DNA vaccines against Plasmodium falciparum].

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.

[Reparative and therapeutic effect of rhBMP-2m on mice injured by chemotherapy].

Aim: To probe the therapeutic effect of recombinant human bone marrow morphogenetic protein-2 maturation peptide(rhBMP-2m) on mouse bone marrow injury caused by cyclophosphamide (CTX).

Methods: 18 mice were divided randomly into 3 groups, namely CTX injection group(CTX group), BMP therapy group(BMP group) and PBS control group(Control group), 6 mice each group. CTX of 200 mg/kg per mouse was intraperitonealy(IP) injected at a time to BMP group and CTX group so as to establish the experimental model of mouse bone marrow injury.

[Construction and expression of prokaryotic expression vector for pTAT-HBV targeted ribonuclease fusion protein].

Aim: To construct Tat-HBV targeted ribonuclease fusion protein prokaryotic expression vector and express it in E.coli.

Methods: The cDNAs encoding HBV targeted ribonuclease, human eosinophil-derived neurotoxin and HBV core protein were respectively cloned into prokaryotic expression vector pTAT-HA.

[Protective antibody response to recombinant fragments of Plasmodium falciparum apical membrane antigen 1].

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.

[Passage adaptability of candidate strains for hemorrhagic fever with renal syndrome purified vaccine in Vero cells and their immunogenicity].

Objective: To adapt the candidate strains of hemorrhagic fever with renal syndrome (HFRS) purified vaccine to Vero cells and to study their antigenicity and immunogenicity.

Methods: The viral strains H8207 (Hantaan virus, HTN) and Y86013 (Seoul virus, SEO) were continuously propagated in Vero cell by the terminal dilution method and studied the characteristics of virus multiplication, viral titers and the amounts of virus antigen after serial passages. Three batches of crude monovalent inactivated vaccine were developed using the different passages of these 2 viral strains.

Anti-HBV effect of TAT- HBV targeted ribonuclease.

Aim: To prepare and purify TAT-HBV targeted ribonuclease fusion protein, evaluate its transduction activity and investigate its effect on HBV replication in 2.2.15 cells.

Inhibition of HBV targeted ribonuclease enhanced by introduction of linker.

Aim: To construct human eosinophil-derived neurotoxin(hEDN) and HBV core protein (HBVc) eukaryotic fusion expression vector with a linker (Gly(4)Ser) (3) between them to optimize the molecule folding, which will be used to inhibit HBV replication in vitro.

Methods: Previously constructed pcDNA3.1(-)/TR was used as a template.